GENE CLUSTER FOR BIOSYNTHESIS OF PAMAMYCIN

This application claims priority of applications with number EP 13198688.7, EP 14169265.7 and EP 14183171.9, all of which are incorporated by reference in their entirety.

FIELD OF THE INVENTION

The invention pertains to the field of production of natural products and, in particular, in the field of production of pamamycin. It provides polynucleotides and polypeptides involved in the biosynthesis of pamamycin and pamamycin precursors as well as vectors and recombinant microorganisms comprising such polynucleotides and polypeptides. Also provided are methods for the production of pamamycins, in particular pamamycin 607 and pamamycin 621 , and their precursors.

BACKGROUND OF THE INVENTION

Pamamycins are a group of macrolide compounds and have been described as early as 1979 (McCann and Pogell, JOURNAL OF ANTIBIOTICS (1979) Vol.: 32(7) pages: 673 to 678). They are produced by several Streptomyces species and have raised scientific interest, because they stimulate the formation of aerial mycelium in Streptomycete species which produce pamamycins, as well as in other streptomycetes which are not known to produce pamamycins. They also enhance the production of secondary metabolites in several Streptomycete species and have antibiotic activity against gram-positive bacteria, fungi and mycobacteria (Pogell, BM ; CELLULAR AND MOLECULAR BIOLOGY (1998); 44(3); pages 461 to 463; Natsume, M. et al. JOURNAL OF ANTIBIOTICS (1995)

Vol.:48(10); pages: 1 159 to 1 164; Hashimoto et al. in BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY. 2003, Vol 67(4), pages 803 to 808 and Hashimoto, M. et a.

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY (201 1 ) Vol. :75(9) pages: 1722 to 1726). Further investigations showed, that the different side chains located on the ring structure of pamamycins, as well as their dimethylamino group have different effects on the capacity of pamamycins to induce aerial mycelia, to inhibit growth of Streptomyces albonigeror to work as antibiotic on Bacillus subtilis, see Natsume, M. et al. JOURNAL OF ANTIBIOTICS (1995) Vol.:48(10); pages:1 159 to 1 164 and Natsume, M. et al.

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY (1995) Vol.:59(9) pages: 1766 to 1768.

The use of C-13- and N-15-labeled precursor units provided further information on the metabolic pathway leading to the production of pamamycins. N-15-labeling suggests that the nitrogen atom of the dimethylamino group present in many pamamycins was derived from the alpha-amino group of an amino acid which has been introduced into the pamaycin structure by transamination, followed by N-methylation. Feeding experiments with C-13- labeled acetate or propionate indicates that the carbon skeleton of pamamycins is derived from six acetate, four propionate and three succinate units, see Hashimoto, M. et al.

JOURNAL OF ANTIBIOTICS (2005) Vol.:58(1 1 ); pages:722 to 730 and Hashimoto, M. et al. BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY (2005) Vol.:69(2) pages:315 to The attractive features and potential uses of pamacins have spured attempts to provide sufficient amounts of pamamycins to enable technical applications. Thus, numerous methods to chemically synthesize pamamycins have been developed and are disclosed for example in Fischer, P et al. CHEMISTRY-A EUROPEAN JOURNAL (201 1) Vol. : 17(47) pages: 13334 to 13340; Germay, O et al. ORGANIC & BIOMOLECULAR CHEMISTRY (2012) Vol.: 10(48) pages: 9709 to 9733; Ren, GB et al. JOURNAL OF ORGANIC

CHEMISTRY (2010) Vol.:75(15) pages: 5048 to 5064; Ren, GB et al. ORGANIC LETTERS (2009) Vol.: 1 1 (24) pages: 5638 to 5641 and Miura, A et al. EUROPEAN JOURNAL OF ORGANIC CHEMISTRY (2008) lssue:29, pages: 4955 to 4962. However, the complex structure of pamamycins require complex and lenthy synthesis methods.

Alternative approaches have tried to avoid this and relied on the use of natural producers of pamamycins. Organisms which produce pamamycins, as well as fermentation methods for the production of pamamycins and methods to purify pamamycins have been disclosed for example in US4283391 , JP62135476A, DE4134168, DE4316836, Natsume, M. et al.

JOURNAL OF ANTIBIOTICS (1995) Vol.:48(10); pages:1 159 to 1 164 and HAERTL et al. THE JOURNAL OF ANTIBIOTICS (1998) VOL. 51 , NO. 1 1 , pp. 1040-1046. These fermentative methods have also been modified to include steps of chemical modification of the produced pamamycins. One important modification has been the hydrolysation of the produced pamamycins in order to produce the antibiotic I MET 4570 which can be used as antibiotic, fungicide and as anionophore, as disclosed in DE4134168. Another fragment of pamamycin produced via hydrolysation is tetrahydorfuranyl-propionic acid, which can be used to transport organic cations into cells and to enhance the availability of cationic therapeutics. Further information can be found in DE4316836. However, the available methods to produce pamamycins via use of their natural producers are limited by the natural production level of these organism. Thus, there is still a need to develop further methods for the production of pamamycins.

Accordingly, the technical problem underlying the present invention can be seen as the provision of additional means and methods for the production of pamamycins, in particular pamamycin 607 and pamamycin 621 . The technical problem is solved by the embodiments characterized in the claims and herein below.

SUMMARY OF THE INVENTION

The invention provides for polypeptides and polynucleotides as well as gene clusters, expression cassettes and vectors comprising one or several of those polynucleotides for the production of pamamycins, which can be used to construct, identify and improve

microorganisms having the capacity to produce one or several pamamycins, in particular to produce pamamycin 607 and/or pamamycin 621. The provided polypeptides have different activities and can be used in different combinations to enhance, or down regulate the capacity of microorganisms to produce one or several pamamycins.

Accordingly, the invention encompasses recombinant microorganism comprising: a) at least one polypeptide having pamC activity, and b) at least one polypeptide having pamG activity, and c) at least one polypeptide having pamF activity, and d) at least one polypeptide having pamA activity, and e) at least one polypeptide having pamB activity, and f) at least one polypeptide having pamD activity, and g) at least one polypeptide having pamE activity, and h) at least one polypeptide having pamO activity, and i) at least one polypeptide having pamK activity, and j) at least one polypeptide having pamJ activity, and k) at least one polypeptide having pamM activity, and I) at least one polypeptide having pamN activity, and m) at least one polypeptide having pamL activity, and n) at least one polypeptide having pamX activity, and o) at least one polypeptide having pamY activity, and p) at least one polypeptide having pamS activity, wherein at least one of those activities a) to p) is up- regulated, or comprising the activities of a) to p) and comprising q) an up-regulated polypeptide having pamR1 activity, or r) a down-regulated polypeptide having pamR2 activity, or s) an up-regulated polypeptide having pamW activity, or t) a down-regulated polypeptide having pamH activity, or u) a combination of at least two of q), r) s) and t), or v) at least one up-regulated activities of a) to p) and at least one of q) to u).

The information on gene clusters being involved in pamamycin production capacity of microorganisms (SEQ ID NO: 1 and 2) allow to identify new microorganisms suitable to be used for the production of pamamycins and to identify new cluster variants comprised in such microorganisms, as well as to design new variants of these clusters, having new gene orders, gene orientation, gene expression, and/or gene numbers for the encoded

polypeptides. Thus, the invention comprises also recombinant microorganisms comprising i) at least one polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2, ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2 and comprising at least one expression cassette for a) at least one polypeptide having pamC activity, and b) at least one polypeptide having pamG activity, and c) at least one polypeptide having pamF activity, and d) at least one polypeptide having pamA activity, and e) at least one polypeptide having pamB activity, and f) at least one polypeptide having pamD activity, and g) at least one polypeptide having pamE activity, and h) at least one polypeptide having pamO activity, and i) at least one polypeptide having pamK activity, and j) at least one polypeptide having pamJ activity, and k) at least one polypeptide having pamM activity, and I) at least one polypeptide having pamN activity, and m) at least one polypeptide having pamL activity, and n) at least one polypeptide having pamX activity, and o) at least one polypeptide having pamY activity, and p) at least one polypeptide having pamS activity iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of i) or ii), if the fragments are combined. The recombinant microorganisms may also comprise at least one expression cassette for q) a polypeptide having pamR1 activity, or r) a polypeptide having pamR2 activity, or s) a polypeptide having pamW activity, or t) a polypeptide having pamH activity, or u) a polypeptide q), r) s) and t) and at least one expression cassette for a polypeptide of q), r) s) and t), which encodes a different polypeptide than the first expression cassette.

Also comprised are recombinant microorganism as described above comprising i) at least one polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 , ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and comprising at least one expression cassette for a) at least one

polypeptide having pamC activity, and b) at least one polypeptide having pamG activity, and c) at least one polypeptide having pamF activity, and d) at least one polypeptide having pamA activity, and e) at least one polypeptide having pamB activity, and f) at least one polypeptide having pamD activity, and g) at least one polypeptide having pamE activity, and h) at least one polypeptide having pamO activity, and i) at least one polypeptide having pamK activity, and j) at least one polypeptide having pamJ activity, and k) at least one polypeptide having pamM activity, and I) at least one polypeptide having pamN activity, and m) at least one polypeptide having pamL activity, and n) at least one polypeptide having pamX activity, and o) at least one polypeptide having pamY activity, and p) at least one polypeptide having pamS activity, and q) at least one polypeptide having pamR1 activity, and r) at least one polypeptide having pamR2 activity, and s) at least one polypeptide having pamW activity, and t) at least one polypeptide having pamH activity, and iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments comprise functional expression cassettes for one or more of the polypeptides of a) to t) and wherein the fragments cover at least the whole sequence of a polynucleotide of i) or ii), if the fragments are combined.

The invention provides also the possibility to enhance the expression of polypeptides for the production of pamamycins via the overexpression of a polypeptide having pamR1 activity. Accordingly, the invention comprises recombinant microorganisms comprising i) at least one polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2, ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2 and comprising at least one expression cassette for a) at least one polypeptide having pamC activity, and b) at least one polypeptide having pamG activity, and c) at least one polypeptide having pamF activity, and d) at least one polypeptide having pamA activity, and e) at least one polypeptide having pamB activity, and f) at least one polypeptide having pamD activity, and g) at least one polypeptide having pamE activity, and h) at least one polypeptide having pamO activity, and i) at least one polypeptide having pamK activity, and j) at least one polypeptide having pamJ activity, and k) at least one polypeptide having pamM activity, and I) at least one polypeptide having pamN activity, and m) at least one polypeptide having pamL activity, and n) at least one polypeptide having pamX activity, and o) at least one polypeptide having pamY activity, and p) at least one polypeptide having pamS activity iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of i) or ii), if the fragments are combined and comprising a recombinant expression cassette for a polypeptide having pamR1 activity wherein the polypeptide is q 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, or q2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17, 1 18, 1 19, 120 and/or 121 , or q3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15, or q4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 1 15, or q5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 per primer, or q6) a polypeptide of q1 ) and/or q2) and comprising the amino acids K105, K190 and at least one of E 153 and E 154, or q7) a polypeptide of at least two of q1), q2), q3), q4), q5) and q6). The polypeptide having pamR1 activity is preferably expressed via an inducible or constitutive promoter.

The recombinant microorganisms as described above and variants therof will ususally have the capacity to produce pamamycins, in particular pamamycin 607 or pamamycin 621 or both. Accordingly, the recombinant microorganisms can be used in methods to produce at least one pamamycin, so that the invention provides for method for the production of pamamycin comprising the steps of: i) cultivating a recombinant microorganism as described above under conditions which allow for the production of pamamycin by said recombinant microorganism; ii) obtaining produced pamamycin, preferably obtaining pamamycin 607 and/or pamamycin 621. Pamamycins produced by the recombinant microorganisms of the invention can be further modified, e.g. by hydrolyzing in order to produce compounds of Formular IV or Formula V, or both. Thus, the invention comprises methods for the production of compounds of the Formula IV or Formula V comprising the steps of: i) cultivating a recombinant microorganism as described above under conditions which allow for the production of pamamycin by said recombinant microorganism; ii) obtaining produced pamamycin, preferably obtaining pamamycin 607 and/or pamamycin 621 , iii) hydrolysing pamamycin, preferably hydrolysing pamamycin 607 and/or pamamycin 621 and iv) obtaining compounds of Formula IV or obtaining compounds of Formula V or obtaining compounds of Formula IV and Formula V. The produced pamamycins may be obtained from the culture medium (culture broth), or from the produced biomass, or from both. The produced pamamycins or compounds of Formula iV or Formula V may be obtained in the form of their agriculturally or pharmacologically acceptable salts.

The gene clusters provided by the invention comprise promoter and terminator sequences as well as transcription factors (pamR1 and pamR2), which regulate the activity of the provided promoter sequences. The promoters and transcription factors can be used to construct systems for coordinated gene expression in recombinant microorganisms comprising 1 ) at least one expression cassette comprising a promoter being able to provide for gene expression in said microorganism and being operably linkted to q1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, or q2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17, 1 18, 1 19, 120 and/or 121 , or q3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15, or q4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 1 15, or q5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 per primer, or q6) a polynucleotide of q1), q2), q3), q4) and/or q5) being obtainable from a pamamycin producing organism, or q7) a polynucleotide of at least two of q1 ), q2), q3), q4), q5) and q6); and 2) at least one expression cassette comprising a promoter comprising i) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 143, 144 or 145, or ii) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 143, 144 or 145, or iii) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive

polynucleotides of SEQ ID NO: 143, 144 or 145 per primer, or iv) a polynucleotide of at least one of i), ii) or iii) and being obtainable from a pamamycin producing organism, or v) a polynucleotide of at least one of i), ii), iii) or iv) and comprising promoter activity in

Streptomyces alboniger. These systems can be used to coordinate the expression of any polypeptide encoding region operatively linked to at least one of the respective promoters. In one embodiment, the polypeptide encoding regions encode one or several of the polypeptides involved in pamamycin production as described above. The recombinant microorganisms comprising such a system for coordinated gene expression are preferably bacteria, yeasts or filamentous fungi, even more preferred they belong to the genus

Streptomyces, in particular to the species Streptomyces albus or Streptomyces alboniger. The information about polypeptides, polynucleotides and gene clusters involved in the production of pamamycin can be used to identify microorganisms which are potentially capable to produce at least one pamamycin, in particular at least one of pamamycin 607 and 621. Hence, the invention comprises methods to identify pamaycin producing microorganisms or microorganisms potentially capable to produce pamamycin comprising the steps of a) providing a microorganism, b) isolating DNA and/or cDNA from such organism, c) identifying polynucleotide sequences encoding polypeptides having at least one of pamC activity, pam G activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity, pamS activity, by I) hybridizing the isolated DNA or RNA under medium or high stringent conditions with polynucleotide fragments of SEQ ID NO: 1 or 2 or both, or II) performing PCR reactions with PCR-primer suitable to amplify fragments of SEQ ID NO: 1 or 2 or both III) sequencing the isolated DNA and/or RNA and perform sequence comparisons with polynucleotide sequences of fragments of SEQ ID NO: 1 or 2 or both, d) selecting microorganisms which comprise sequences encoding polypeptides for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15 or all of the polynucleotide sequences of d), e) testing the selected microorganisms for production of pamamycin. The invention does also provide for methods to enhance the production of pamamycin, preferably the production of pamamycin 607 or pamamycin 621 or both, in a microorganism comprising i) at least one polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2, preferably 2, ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2, preferably 2, and comprising at least one expression cassette for each one of a) to p) as described above; iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of i) or ii), if the fragments are combined by providing the microorganism with at least one expression cassette comprising a promoter being able to provide for gene expression in said microorganism and being operably linkted to q1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, or q2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17, 1 18, 1 19, 120 and/or 121 , or q3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15, or q4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ I D NO: 1 15, or q5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 per primer, or q6) a polynucleotide of q1), q2), q3), q4) and/or q5) being obtainable from a pamamycin producing organism, or q7) a

polynucleotide of at least two of q 1 ), q2), q3), q4), q5) and q6).

Because of the versatile applications of the polypeptides, polynucleotides, gene clusters, expression cassettes, vectors, host cells, recombinant micororganisms and systems for coordinated gene expression, the invention comprises also the use of one or several of them in any method, in particular in a method involving the production of at least one pamamycin, the identification of microorganisms potentially being able to produce at least one pamamycin, methods to improve the capacity of a microorganism to produce at least one pamamycin.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows schematic drawings of different pamamycin gene clusters. The drawing of Cluster 1 represents a polynucleotide comprising polypeptide encoding regions for polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity, respectively. The drawing of Cluster 2 represents a cluster, which comprises polypeptide encoding regions for all of the activites mentioned for Cluster 1 , but comprises in addition thereto a polypeptide encoding region encoding a polypeptide having pamH activity. The drawing of Cluster 3 represents a cluster comprising polypeptide encoding regions for all activities mentioned for Cluster 1 , but comprises also a polypeptide encoding region encoding a polypeptide having pamR1 activity. The drawing of Cluster 4 represents a cluster comprising polypeptide encoding regions for all activities mentioned for Cluster 1 , but comprises also a polypeptide encoding region encoding a polypeptide having pamH activity and a polypeptide encoding region encoding a polypeptide having pamR1 activity. The drawing of Cluster 5 represents a cluster comprising polypeptide encoding regions for all activities mentioned for Cluster 4, but comprises also a polypeptide encoding region encoding a polypeptide having pamR2 activity and a polypeptide encoding region encoding a polypeptide having pamW activity. The boxed arrows represent the order and orientation of the polypeptide encoding regions, wherein the colour or pattern of the boxed arrows represent the general class or family of the encoded polypeptides based on sequence homology, e.g. the encoded polypeptides having pamG, pamF, pamA, pam D, pam E, pamK and pamJ activity are represendted by black boxed arrows. The legend of boxed arrows provided under the representations of the clusters shows that these polypeptides belong to the homology class or family of ketosynthases " KS" . The polypeptide having pamB activity, represented by a white boxed arrow with black borders belongs to the homology class of acetyltransferases " AT" . The same principle applies to the rest of the listed homology class or families depicted in the legend, wherein " KR/DH" represent ketoreductases/dehydrogenases, " ACP" represent acyl carrier proteins, " hydrolase" represent hydrolases, " AmT" represents aminotransferases, " MT" represents methyltransferases, " transport" represent polypeptides having transporter functions, " enoyl-CoA hydratases" represents enoyl-CoA hydratases, " CoA-ligase" represents CoA-ligases, " regulation" represents polypeptides having regulatory functions. The schematic drawings of the clusters are intended to represent polynucleotides comprising promoter and terminator sequences operatively connected to the polypeptide encoding regions, which are suited to enable expression of the respective polypeptide in a microorganism, preferably of the genus Streptomyces. The schematic drawings of the clusters are intended to represent also polynucleotides comprising at least one of the different polypeptide encoding regions in a different order and/or orientation. A preferred example of Cluster 1 is disclosed as SEQ ID NO: 1 , while a preferred example of Cluster 5 is disclosed as SEQ ID NO: 2. Preferred examples of Cluster 2 and 4 are respective fragments of SEQ ID NO: 2, while a preferred version of Cluster 3 is a fragment of SEQ ID NO: 2, wherein expression of the polypeptide having pamH activity is prevented, e.g. by deletion or mutation of parts or all of the polypeptide encoding region or its promoter, or by substitution of parts or all of the pamD encoding region or expression cassette with a pamR1 encoding region or expression cassette. Examples of Cluster 3 preferably express the polypeptide having pamR1 activity under control of a constitutive promoter. These examples of Clusters 1 , 2, 3, 4 or 5 are preferably used for expression of the endocded polypeptides in microorganisms of the genus Streptomyces, such as, but not excluding others, Streptomyces alboniger and Streptomyces albus.

Figure 2 schows a schematic drawing of the metabolic pathway leading to pamamycins. The first step involves the activity of pamA, followed by the activity of pamB, pamD and the activity of pamE. The pathway splits in order to produce the compound S-chain via the activities of pamO, pamM, pamN, pamS and to procude the compound K-chain via the activities of pamF, pamG, pamO, pamM, pamN and pamS. Which is then further modified to produce the compound L-chain via the activities of pamX and pamY. The produced compounds S-chain and L-chain are combined with Coenzyme A (CoASH) via the activitiy of pamL. The final pamamycins are then produced by the combination of one compound S- chain-SCoA with one compound L-chain-SCoA, mediated by the activity of pamJ and pamK. The residues described as R1 , R2, R3, R4 and R5 of all formulas are the same residues as listed in the definition of Formula (I) given below.

Figure 3 represents an amino acid sequence alignment of the polypeptide (SEQ ID NO: 4) encoded by gene pamC and variants thereof (SEQ ID NO: 5, 6, 7, 8, and 9). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being L29, D32, S33, E44 and F47. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 4a and Figure 4b (together Figure 4) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 1 1) encoded by gene pamG and variants thereof (SEQ ID NO: 12, 13, 14, 15 and 16). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being P19, A64, C1 16, L139, D163, A167, H258, G287 and N288. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background. Amino acid P19 may be substituted by another amino acid in preferred variants, if P20 is not changed. Figures 5a to Figure 5c (together Figure 5) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 18) encoded by gene pamF and variants thereof (SEQ ID NO: 19, 20, 21 , 22 and 23). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being G16, A186, G255, G261 , G366 and H267. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background. Figures 6a to Figure 6c (together Figure 6) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 25) encoded by gene pamA and variants thereof (SEQ ID NO: 26, 27, 28, 29 and 30). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in one or all the amino acids marked with a black arrow, being G21 , C191 , G258, G264, H333, G371 and H272, more preferred, they also have no change in at least one of the amino acids A190, V192, G193, G334, T335 and Y370, e.g. having no change in the amino acids G21 , A190, G258, G264, G371 and H272. More preferred, the variants will only have

conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 7a to Figure 7c (together Figure 7) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 32) encoded by gene pamB and variants thereof (SEQ ID NO: 33, 34, 35, 36 and 37). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being T2, G3, P6, R22, A23, P59, L89, D218, G221 , N222, P260, P266, V270, G407, G409, E434, G436, G491 , G506, G508, G509. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background. One amino acid of G506, G508, G509 may be substituted in the preferred variants, in case the other two are kept identical.

Figures 8a and Figure 8b (together Figure 8) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 39) encoded by gene pamD and variants thereof (SEQ ID NO: 40, 41 , 42, 43 and 44). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being P21 , P101 , C121 , H246, G275 and G308. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background. Figures 9a and Figure 9b (together Figure 9) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 46) encoded by gene pamE and variants thereof (SEQ ID NO: 47, 48, 49, 50 and 51 ). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being P25, P103, C136, H270, G299 and G333. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background. Figures 10a and Figure 10b (together Figure 10) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 53) encoded by gene pamO and variants thereof (SEQ ID NO: 54, 55, 56, 57 and 58). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being G1 1 , G17, D61 , Y164, K168, P194 and at least two of G256, G257 and G258. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 1 1a and Figure 1 1 b (together Figure 1 1) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 60) encoded by gene pamK and variants thereof (SEQ ID NO: 61 , 62, 63, 64 and 65). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being F33, C159, G314, G346 and G348. One of G346 and G348 may be substituted, in case G347 is kept. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background..

Figures 12a to Figure 12c (together Figure 12) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 67) encoded by gene pamJ and variants thereof (SEQ ID NO: 68, 69, 70, 71 and 72). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being G14, G246, G252, G256 and H257. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background. Figures 13a to Figure 13c (together Figure 13) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 74) encoded by gene pamM and variants thereof (SEQ ID NO: 75, 76, 77, 78 and 79). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being G1 1 , V139, G241 , G247, D291 , A319, Y384, K388, P414, G415, T419, G465, G473. G473 may be substituted, in case G472 is kept identical. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background. Figures 14a and Figure 14b (together Figure 14) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 81 ) encoded by gene pamN and variants thereof (SEQ ID NO: 82, 83, 84, 85 and 86). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being G16, G22, D66, Y158, K162, P188 and G189. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 15a to Figure 15c (together Figure 15) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 88) encoded by gene pamL and variants thereof (SEQ ID NO: 89, 90, 91 , 92 and 93). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being L82, V93, T214, S215, G216, T218, G219, K222, E352, G370, G394, Y410, T438, G439, D430, R445, G454, E463, P520, P530, G535 and K536. T218 may be substituted, in case T217 is kept. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 16a to Figure 16c (together Figure 16) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 95) encoded by gene pamX and variants thereof (SEQ ID NO: 96, 97, 98, 99 and 100). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being G41 , D46, G57, G1 13, H147, D241 , K271 , G276 and T308. G41 may be substituted, in case G40 is kept. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 17a and Figure 17b (together Figure 17) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 102) encoded by gene pamY and variants thereof (SEQ ID NO: 103, 104, 105, 106 and 107). The amino acids are

represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being G67, G69, D86, G154 and G244. G154 and/or G244 may be substituted, in case the neighboring G153 and G243 are kept. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 18a and Figure 18b (together Figure 18) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 109) encoded by gene pamS and variants thereof (SEQ ID NO: 1 10, 1 1 1 , 1 12, 1 13 and 1 14). The amino acids are

represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being N41 , F76, G79, D81 , G1 16, G120, G122, D130, G148, G187 and K221. G120 or G122 may be substituted, in case G121 is kept. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 19a and Figure 19b (together Figure 19) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 1 16) encoded by gene pamR1 and variants thereof (SEQ ID NO: 1 17, 1 18, 1 19, 120 and 121 ). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being K105, E153 and K190. E153 may be substituted, in case E154 is kept. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 20a and Figure 20b (together Figure 20) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 123) encoded by gene pamH and variants thereof (SEQ ID NO: 124, 125, 126, 127 and 128). The amino acids are

represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being D16, N1 17, H193, P1 19, H227 and N317. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background. Figures 21a and Figure 21 b (together Figure 21) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ I D NO: 130) encoded by gene pamR2 and variants thereof (SEQ ID NO: 131 , 132, 133, 134 and 135). The amino acids are represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being E3, W19, A41 , G49, A59, Y70, L79, G169, P207, F226, L230 and G236. A41 may be substituted, in case A40 is kept. G49 may be substituted, in case G48 is kept. L79 may be substituted, in case L80 is kept. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 22a to Figure 22c (together Figure 22) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ I D NO: 137) encoded by gene pamW and variants thereof (SEQ ID NO: 138, 139, 140, 141 and 142). The amino acids are

represented according to the standard single letter amino acid code. Further sequence variants can be deduced from this alignment. Such sequence variants preferably have no change in the amino acids marked with a black arrow, being D92, G95 and G131. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background.

Figures 23a to Figure 23c (together Figure 23) represent consecutive parts of an amino acid sequence alignment of polypeptides (SEQ ID NO: 349, 356, 363, 370 and 377, see also Table 8g) which show pamB activity and which are comprised by different

Streptomyces species. The amino acids are represented according to the standard single letter amino acid code. Examples of sequence variants are described by SEQ ID NO: 349, 356, 363, 370, 377). Further sequence variants can be deduced from the alignment in Figure 23, as well as from alignments of SEQ ID NO: 349 to 354, 356 to 361 , 363 to 368, 370 to 375 and 377 to 382). Such sequence variants preferably have no change in the amino acids marked with a black arrow in Figure 23, being G34, P46, L77, T146, P162, D188, G191 , P230, P226, V240, G369, G, 371 , E396, G471 and R586. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background in Figure 23.

Figures 24a to Figure 24b (together Figure 24) represent consecutive parts of an amino acid sequence alignment of the polypeptide (SEQ ID NO: 384, 391 , 398, 405 and 412, see also Table 1 1g) which show pamO activity and which are comprised by different

Streptomyces species. The amino acids are represented according to the standard single letter amino acid code. Examples of sequence variants are described by SEQ ID NO: 384, 391 , 398, 405 and 412). Further sequence variants can be deduced from the alignment in Figure 24, as well as from alignments of SEQ ID NOs: 284 to 389, 391 to 396, 398 to 403, 405 to 410 and 412 to 417). Such sequence variants preferably have no change in the amino acids marked with a black arrow in Figure 24, being G22, G28, D72, Y177, K181 , and P207. More preferred, the variants will only have conservative amino acid substitutions in the amino acids depicted as white letters on a black background in Figure 24.

Figure 25a depicts results of Example 4. The production of pamamycin 607 of the natural pamamycin producing organism Streptomyces alboniger DSM 40043 and of a recombinant strain of Streptomyces albus J 1074 is confirmed by testing the SGG culture medium

(" Super" ) and the produced biomass (" Biomass" ) for the presence of pamamycin. The recombinant strain of Streptomyces albus J 1074 comprises the R2 cosmid with the pamamycin cluster depicted in Figure 1 as Cluster 5. It is shown that the recombinant strain of Streptomyces albus J 1074 (" S. albus + R22" ) comprises less pamamycin in the produced biomass, but more pamamycin in the culture medium than the wildtype strain of Streptomyces albus J 1074.

Figure 25b depicts results of Example 5. Three different strains of pamamycin producing microorganisms are analysed for their production of pamamycin 607 in two different media, and separately analysed for pamamycin 607 production in the culture supernatant (" PPM KF" and (" SGG KF) or in the produced biomass (" PPM Mycel" and " SGG Mycel" ) of the respective media PPM and SGG described in Example 1. The first strain is Streptomyces albus J 1074 comprising the R2 cosmid (" S. albus + R2" ), the second strain is the wildtype of Streptomyces alboniger DSM 40043 (" S. alboniger" ), while the third strain is a recombinant Streptomyces alboniger DSM 40043 comprising the R2 cosmid (" S. alboniger + R2" ). The overall production of pamamycin of each strain is depicted for the medium PPM and SSG (" PPM" and " SGG" ). This highest overall production of pamamycin is achieved with a recombinant Streptomyces alboniger DSM 40043 comprising the R2 cosmid.

Figure 26 depicts results of Example 8. Four different strains of Streptomyces alboniger. The first strain is the wildtype of Streptomyces alboniger DSM 40043 (" S. alboniger" ), the second strain is a recombinant Streptomyces alboniger DSM 40043 comprising the R2 cosmid (" S. alboniger + R2" ), the third strain is a recombinant Streptomyces alboniger DSM 40043 (" pTOSR1 P21 " ) overexpressing the pamR1 polypeptide from the

polypeptide encoding region (SEQ ID NO: 129) obtained from Streptomyces alboniger DSM 40043 under transcriptional control of the heterologous p21 promoter (SEQ ID NO: 193), while the fourth strain is a recombinant Streptomyces alboniger DSM 40043 overexpressing the pamR1 polypeptide from the polypeptide encoding region (SEQ ID NO: 129) obtained of Streptomyces alboniger DSM 40043 (" pTOSRI BX" ) under

transcriptional control of its homolgous pamR1 promoter (SEQ ID NO: 145). All four strains are grown in the culture medium " PPM" or " SGG" , respectively. Figure 26 shows that all four strains produce pamamycin 607, but on different level. The highest level of production is obtained with the fourth strain.

Table 1 :

Pamamycin R1 R2 R3 R4 R5

593 CH ₃ H CHs CHs H

607 CHs CHs CHs CHs H

621A CHs CHs CHs CHs CHs

621 B CH2CH5 H CHs CHs CHs

Formula (I)

The term " pamamycin 593" means a compound of Formula (I), having the respective residues R1 , R2, R3, R4 and R5 as defined in Table 1.

The term " pamamycin 607" means a compound of Formula (I), having the respective residues R1 , R2, R3, R4 and R5 as defined in Table 1. The term " pamamycin 621 " means compounds of Formula (I), having the respective residues R1 , R2, R3, R4 and R5 for 621 A, 621 B, 621 C, or 621 D as defined in Table 1. Usually the term pamamycin 621 refers to a mixture of at least two of 621 A, 621 B, 621 C and 621 D. The terms " pamamycin 621 A" , " pamamycin 621 B" , " pamamycin

621 C" and " pamamycin 621 D" mean compounds of Formula (I), having the respective residues R1 , R2, R3, R4 and R5 for 621 A, 621 B, 621 C, or 621 D as defined in Table 1.

The term " pamamycin 635" means compounds of Formula (I), having the respective residues R1 , R2, R3, R4 and R5 for 635A, 635B, 635C, 635D, 635E, or 635F as defined in Table 1. Usually the term pamamycin 635 refers to a mixture of at least two of 635A, 635B, 635C, 635D, 635E, and 635F. The terms " pamamycin 635A" , " pamamycin 635B" , " pamamycin 635C" , " pamamycin 635D" , " pamamycin 635E" and " pamamycin 35F" mean compounds of Formula (I), having the respective residues R1 , R2, R3, R4 and R5 for 621 A, 621 B, 621C, 621 D, 621 E or 621 F as defined in Table 1. The term " pamamycin 649" means a mixture of compounds of Formula (I), having the respective residues R1 , R2, R3, R4 and R5 for 649A or 649B as defined in Table 1. The terms " pamamycin 649A" and " pamamycin 649B" mean compounds of Formula (I), having the respective residues R1 , R2, R3, R4 and R5 for 649A, or 6249B as defined in Table 1.

The term " S-chain" means compounds of Formula (II) AND Enantiomer

Formula (II)

R4 and R5 can be H, CH ₃ or CH2CH5 in combinations as shown for R4 and R5 in Table 1. SCoA, representing bound Coenzyme A, can also be OH, preferably it is OH.

The term " L-chain" means a compound of Formula (III),

AND Enantiomer

Formula (III)

R1 , R2 and R3 can be H, CH3 or CH2CH5 in combinations as shown for R1 , R2 and R3 in Table 1. SCoA, representing bound Coenzyme A, can also be OH, preferably it is OH.

The term " K-chain" means a compound of Formula (IV),

The term„ IMET4570" means a compound of Formula (V)

C H ₃ C H ₃

Formula (V) in which

R1 is hydrogen, a carboxyl, hydroxyl, ester, amide or hydroxamate group, a halogenide or a salt with anorganic or organic catione, and

R2 is hydrogen, a hydroxyl, ester or ether group or a halogenide,

or wherein the compound of Formula (iV) is a salt of the dimethylamino group.

The term„ Tetrahydrofuranylpropionic acid" means compounds of Formula (VI)

Variant I: R1 =CH ₃ , R2= H;

Variant II: R1 and R2 = H;

Variant III: R1 = CH ₃ ; R2: = COCH ₃ )

Formula (VI)

The term " about" is used herein to mean approximately, roughly, around, or in the region of. When the term " about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values-set forth. In general, the term " about" is used herein to modify a numerical value above and below the stated value by a variance of 20 percent up or down (higher or lower), preferably 15 percent, more preferably 10 percent and most preferably 5 percent.

The term "genome" or "genomic DNA" is referring to the heritable genetic information of a host organism. Said genomic DNA comprises the entire genetic material of a cell or an organism, including the DNA of the nucleus (chromosomal DNA), extrachromosomal DNA, and organellar DNA (e.g. of mitochondria). Preferably, the terms genome or genomic DNA is referring to the chromosomal DNA of the nucleus.

The term "chromosomal DNA" or "chromosomal DNA sequence" is to be understood as the genomic DNA of the cellular nucleus independent from the cell cycle status. Chromosomal DNA might therefore be organized in chromosomes or chromatids, they might be

condensed or uncoiled. An insertion into the chromosomal DNA can be demonstrated and analyzed by various methods known in the art like e.g., polymerase chain reaction (PCR) analysis, Southern blot analysis, fluorescence in situ hybridization (FISH), in situ PCR and next generation sequencing (NGS).

The term " promoter" refers to a polynucleotide which directs the transcription of a structural gene to produce mRNA. Typically, a promoter is located in the 5' region of a gene/operon, proximal to the start codon of a structural gene. If a promoter is an inducible promoter, then the rate of transcription increases in response to an inducing agent. In contrast, the rate of transcription is not regulated by an inducing agent, if the promoter is a constitutive promoter.

The term " enhancer" refers to a polynucleotide. An enhancer can increase the efficiency with which a particular gene is transcribed into mRNA irrespective of the distance or orientation of the enhancer relative to the start site of transcription. Usually an enhancer is located close to a promoter, a 5' -untranslated sequence or in an intron.

A polynucleotide is "heterologous to" an organism or a second polynucleotide if it originates from a foreign species, or, if from the same species, is modified from its original form. For example, a promoter operably linked to a heterologous coding sequence refers to a coding sequence from a species different from that from which the promoter was derived, or, if from the same species, a coding sequence which is not naturally associated with the promoter (e. g. a genetically engineered coding sequence or an allele from a different ecotype or variety).

"Transgene", " transgenic" or "recombinant" refers to a polynucleotide manipulated by man or a copy or complement of a polynucleotide manipulated by man. For instance, a transgenic expression cassette comprising a promoter operably linked to a second polynucleotide may include a promoter that is heterologous to the second polynucleotide as the result of manipulation by man (e.g., by methods described in Sambrook et al., Molecular Cloning-A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, (1989) or Current Protocols in Molecular Biology Volumes 1 -3, John Wiley & Sons, Inc. (1994-1998)) of an isolated nucleic acid comprising the expression cassette. In another example, a recombinant expression cassette may comprise polynucleotides combined in such a way that the polynucleotides are extremely unlikely to be found in nature. For instance, restriction sites or plasmid vector sequences manipulated by man may flank or separate the promoter from the second polynucleotide. One of skill will recognize that polynucleotides can be manipulated in many ways and are not limited to the examples above.

In case the term "recombinant" is used to specify an organism or cell, e.g. a microorganism, it is used to express that the organism or cell comprises at least one "transgene",

" transgenic" or "recombinant" polynucleotide, which is usually specified later on. The term "recombinant" comprises also " man-made mutations" or " man-made mutants" Both are not isolated from nature, but are the result of artificial mutagenesis and/or selection pressure created by interference from man. Usually such mutations are produced by exposing the respective organisms to mutagenic chemicals or mutagenic irradiation . Man-made mutants are microorganisms comprising man-made mutations. A polynucleotide "exogenous to" an individual organism is a polynucleotide which is introduced into the organism by any means other than by a sexual cross.

The terms " operable linkage" or " operably linked" are generally understood as meaning an arrangement in which a genetic control sequence, e.g. a promoter, enhancer or terminator, is capable of exerting its function with regard to a polynucletide being operably linked to it, for example a polynucleotide encoding a polypeptide. Function, in this context, may mean for example control of the expression, i.e. transcription and/or translation, of the nucleic acid sequence. Control, in this context, encompasses for example initiating, increasing, governing or suppressing the expression, i.e. transcription and, if appropriate, translation. Controlling, in turn, may be, for example, growth stage- and / or cell type- specific. It may also be inducible, for example by certain chemicals, stress, pathogens and the like. Preferably, operable linkage is understood as meaning for example the sequential arrangement of a promoter, of the nucleic acid sequence to be expressed and, if

appropriate, further regulatory elements such as, for example, a terminator, in such a way that each of the regulatory elements can fulfill its function when the nucleic acid sequence is expressed. An operably linkage does not necessarily require a direct linkage in the chemical sense. Genetic control sequences such as, for example, enhancer sequences are also capable of exerting their function on the target sequence from positions located at a distance to the polynucleotide, which is operably linked. Preferred arrangements are those in which the nucleic acid sequence to be expressed is positioned after a sequence acting as promoter so that the two sequences are linked covalently to one another. The distance between the promoter and the amino acid sequence encoding polynucleotide in an expression cassette, is preferably less than 200 base pairs, especially preferably less than 100 base pairs, very especially preferably less than 50 base pairs. The skilled worker is familiar with a variety of ways in order to obtain such an expression cassette. However, an expression cassette may also be constructed in such a way that the nucleic acid sequence to be expressed is brought under the control of an endogenous genetic control element, for example an endogenous promoter, for example by means of homologous recombination or else by random insertion. Such constructs are likewise understood as being expression cassettes for the purposes of the invention. The term " expression cassette" means those construct in which the nucleic acid sequence encoding an amino acid sequence to be expressed is linked operably to at least one genetic control element which enables or regulates its expression (i.e. transcription and / or translation). The expression may be, for example, stable or transient, constitutive or inducible.

The terms " express," " expressing," " expressed" and " expression" refer to expression of a gene product (e.g., a biosynthetic enzyme of a gene of a pathway or reaction defined and described in this application) at a level that the resulting enzyme activity of this protein encoded for, or the pathway or reaction that it refers to allows metabolic flux through this pathway or reaction in the organism in which this gene/pathway is expressed. The expression can be done by genetic alteration of the microorganism that is used as a starting organism. In some embodiments, a microorganism can be genetically altered (e.g., genetically engineered) to express a gene product at an increased level relative to that produced by the starting microorganism or in a comparable microorganism which has not been altered. Genetic alteration includes, but is not limited to, altering or modifying regulatory sequences or sites associated with expression of a particular gene (e.g. by adding strong promoters, inducible promoters or multiple promoters or by removing regulatory sequences such that expression is constitutive), modifying the chromosomal location of a particular gene, altering nucleic acid sequences adjacent to a particular gene such as a ribosome binding site or transcription terminator, increasing the copy number of a particular gene, modifying proteins (e.g., regulatory proteins, suppressors, enhancers, transcriptional activators and the like) involved in transcription of a particular gene and/or translation of a particular gene product, or any other conventional means of deregulating expression of a particular gene using routine in the art (including but not limited to use of antisense nucleic acid molecules, for example, to block expression of repressor proteins).

In some embodiments, a microorganism can be physically or environmentally altered to express a gene product at an increased or lower level relative to level of expression of the gene product unaltered microorganism. For example, a microorganism can be treated with, or cultured in the presence of an agent known, or suspected to increase transcription of a particular gene and/or translation of a particular gene product such that transcription and/or translation are enhanced or increased. Alternatively, a microorganism can be cultured at a temperature selected to increase transcription of a particular gene and/or translation of a particular gene product such that transcription and/or translation are enhanced or increased.

The terms " deregulate," " deregulated" and " deregulation" refer to alteration or modification of at least one gene in a microorganism, wherein the alteration or modification results in increasing efficiency of production of a given compound in the microorganism relative to production in absence of the alteration or modification. In some embodiments, a gene that is altered or modified encodes an enzyme in a biosynthetic pathway, or a transport protein, such that the level or activity of the biosynthetic enzyme in the

microorganism is altered or modified, or that the transport specificity or efficiency is altered or modified. In some embodiments, at least one gene that encodes an enzyme in a biosynthetic pathway, i.e. a polypeptide bringing about a specific activity in the biosynthetic pathway, is altered or modified such that the level or activity of the enzyme is enhanced or increased relative to the level in presence of the unaltered or wild type gene.

Deregulation also includes altering the coding region of one or more genes to yield, for example, an enzyme that is feedback resistant or has a higher or lower specific activity. Also, deregulation further encompasses genetic alteration of genes encoding transcriptional factors (e.g., activators, repressors) which regulate expression of genes coding for enzymes or transport proteins. The terms " deregulate," " deregulated" and " deregulation" can further be specified in regard to the kind of deregulation present.

In case the particular activity, is altered or modified such that the level or activity of the enzyme is enhanced or increased relative to the level in presence of the unaltered or wild type gene, the term " up-regulated" is used. In case particular activity, is altered or modified such that the level or activity of the enzyme is lowered or decreased relative to the level in presence of the unaltered or wild type gene, the term " down -regulated" is used. Preferably the recombinant microorganism having an " up-regulated" activity comprises additional expression cassettes for the expression of polypeptides having the respective up- regulated activity and/or may have an modified or exchanged promoter region for the respective endogenous expression cassette, in order to produce more of the polypeptide. However, the activities may be also up-regulated according to other technologies known to a person skilled in the art, for example but not excluding others, modifiying the mRNA sequence in order to enhance its stability or promote its translation, by using a codon optimized mRNA sequence, or by down-regulating an repressor of the respective activity or expression of the polypeptide providing the respective activity.

Similar means, but having the opposite effects, can be applied in order to " down- regulate" a particular activity.

The term " deregulated" includes expression of a gene product at a level lower or higher than that expressed prior to manipulation of the microorganism or in a comparable microorganism which has not been manipulated. In one embodiment, the microorganism can be genetically manipulated (e.g., genetically engineered) to express a level of gene product at a lesser or higher level than that expressed prior to manipulation of the microorganism or in a comparable microorganism which has not been manipulated. Genetic manipulation can include, but is not limited to, altering or modifying regulatory sequences or sites associated with expression of a particular gene (e.g., by removing strong promoters, inducible promoters or multiple promoters), modifying the chromosomal location of a particular gene, altering nucleic acid sequences adjacent to a particular gene such as a ribosome binding site or transcription terminator, decreasing the copy number of a particular gene, modifying proteins (e.g., regulatory proteins, suppressors, enhancers, transcriptional activators and the like) involved in transcription of a particular gene and/or translation of a particular gene product, or any other conventional means of deregulating expression of a particular gene routine in the art (including but not limited to use of antisense nucleic acid molecules, or other methods to knock-out or block expression of the target protein).

The term " deregulated gene activity" also means that a gene activity is introduced into a microorganism where the respective gene activity, has not been observed before, e.g. by introducing a recombinant gene, e.g. a heterologous gene, in one or more copies into the microorganism preferably by means of genetic engineering.

The phrase " deregulated pathway or reaction" refers to a biosynthetic pathway or reaction in which at least one gene that encodes an enzyme in a biosynthetic pathway or reaction is altered or modified such that the level or activity of at least one biosynthetic enzyme is altered or modified. The phrase " deregulated pathway" includes a biosynthetic pathway in which more than one gene has been altered or modified, thereby altering level and/or activity of the corresponding gene products/enzymes. In some cases the ability to " deregulate" a pathway (e.g., to simultaneously deregulate more than one gene in a given biosynthetic pathway) in a microorganism arises from the particular phenomenon of microorganisms in which more than one enzyme (e.g., two or three biosynthetic enzymes) are encoded by genes occurring adjacent to one another on a contiguous piece of genetic material termed a " cluster" or " gene cluster" In other cases, in order to deregulate a pathway, a number of genes must be deregulated in a series of sequential engineering steps.

To express the deregulated genes according to the invention, the DNA sequence encoding the polypeptide must be operably linked to regulatory sequences that control transcriptional expression in an expression vector and then, introduced into either microorganism. In addition to transcriptional regulatory sequences, such as promoters and enhancers, expression vectors can include translational regulatory sequences and a marker gene which is suitable for selection of cells that carry the expression vector.

The terms " overexpress" , " overexpressing" , " overexpressed" and

" overexpression" refer to expression of a gene product, in particular to enhancing the expression of a gene product at a level greater than that present prior to a genetic alteration of the starting microorganism. In some embodiments, a microorganism can be genetically altered (e.g., genetically engineered) to express a gene product at an increased level relative to that produced by the starting microorganism. Genetic alteration includes, but is not limited to, altering or modifying regulatory sequences or sites associated with expression of a particular gene (e.g., by adding strong promoters, inducible promoters or multiple promoters or by removing regulatory sequences such that expression is constitutive), modifying the chromosomal location of a particular gene, altering nucleic acid sequences adjacent to a particular gene such as a ribosome binding site or transcription terminator, increasing the copy number of a particular gene, modifying proteins (e.g., regulatory proteins, suppressors, enhancers, transcriptional activators and the like) involved in transcription of a particular gene and/or translation of a particular gene product, or any other conventional means of deregulating expression of a particular gene using routine in the art (including but not limited to use of antisense nucleic acid molecules, for example, to block expression of repressor proteins). Another way to overexpress a gene product is to enhance the stability of the gene product to increase its life time.

The term "domain" refers to a set of amino acids conserved at specific positions along an alignment of sequences of evolutionarily related proteins. While amino acids at other positions can vary between homologues, amino acids that are highly conserved at specific positions indicate amino acids that are likely essential in the structure, stability or function of a protein. Identified by their high degree of conservation in aligned sequences of a family of protein homologues, they can be used as identifiers to determine if any polypeptide in question belongs to a previously identified polypeptide family.

The term " motif" or " consensus sequence" or " signature" refers to a short conserved region in the sequence of evolutionarily related proteins. Motifs are frequently highly conserved parts of domains, but may also include only part of the domain, or be located outside of conserved domain (if all of the amino acids of the motif fall outside of a defined domain). Specialist databases exist for the identification of domains, for example, SMART (Schultz et al. (1998) Proc. Natl. Acad. Sci. USA 95, 5857-5864; Letunic et al. (2002) Nucleic Acids Res 30, 242-244), InterPro (Mulder et al., (2003) Nucl. Acids. Res. 31 , 315-318), Prosite (Bucher and Bairoch (1994), A generalized profile syntax for biomolecular sequences motifs and its function in automatic sequence interpretation. (In) ISMB-94; Proceedings 2nd International Conference on Intelligent Systems for Molecular Biology. Altman R., Brutlag D., Karp P., Lathrop R., Searls D., Eds., pp53-61 , AAAI Press, Menlo Park; Hulo et al., Nucl. Acids. Res. 32:D134-D137, (2004)), or Pfam (Bateman et al., Nucleic Acids Research 30(1 ): 276-280 (2002) & The Pfam protein families database: R.D. Finn, J. Mistry, J. Tate, P. Coggill, A. Heger, J.E. Pollington, O.L. Gavin, P. Gunesekaran, G. Ceric, K. Forslund, L. Holm, E.L. Sonnhammer, S.R. Eddy, A. Bateman Nucleic Acids Research (2010) Database Issue 38:D21 1 -222). A set of tools for in silico analysis of protein sequences is available on the ExPASy proteomics server (Swiss Institute of Bioinformatics (Gasteiger et al., ExPASy: the proteomics server for in-depth protein knowledge and analysis, Nucleic Acids Res. 31 :3784-3788(2003)). Domains or motifs may also be identified using routine techniques, such as by sequence alignment.

Methods for the alignment of sequences for comparison are well known in the art, such methods include GAP, BESTFIT, BLAST, FASTA and TFASTA. GAP uses the algorithm of Needleman and Wunsch ((1970) J Mol Biol 48: 443-453) to find the global (i.e. spanning the complete sequences) alignment of two sequences that maximizes the number of matches and minimizes the number of gaps. The BLAST algorithm (Altschul et al. (1990) J Mol Biol 215: 403-10) calculates percent sequence identity and performs a statistical analysis of the similarity between the two sequences. The software for performing BLAST analysis is publicly available through the National Centre for Biotechnology Information (NCBI).

Homologues may readily be identified using, for example, the ClustalW multiple sequence alignment algorithm (version 1.83), with the default pairwise alignment parameters, and a scoring method in percentage. Global percentages of similarity and identity may also be determined using one of the methods available in the MatGAT software package

(Campanella et al., BMC Bioinformatics. 2003 Jul 10;4:29. MatGAT: an application that generates similarity/identity matrices using protein or DNA sequences.). Minor manual editing may be performed to optimize alignment between conserved motifs, as would be apparent to a person skilled in the art. Furthermore, instead of using full-length sequences for the identification of homologues, specific domains may also be used. The sequence identity values may be determined over the entire nucleic acid or amino acid sequence or over selected domains or conserved motif(s), using the programs mentioned above using the default parameters. For local alignments, the Smith-Waterman algorithm is particularly useful (Smith TF, Waterman MS (1981) J. Mol. Biol 147(1); 195-7).

Typically, this involves a first BLAST involving BLASTing a query sequence against any sequence database, such as the publicly available NCBI database. BLASTN or TBLASTX (using standard default values) are generally used when starting from a nucleotide sequence, and BLASTP or TBLASTN (using standard default values) when starting from a protein sequence. The BLAST results may optionally be filtered. The full-length sequences of either the filtered results or non-filtered results are then BLASTed back (second BLAST) against sequences from the organism from which the query sequence is derived. The results of the first and second BLASTS are then compared. A paralogue is identified if a high-ranking hit from the first blast is from the same species as from which the query sequence is derived, a BLAST back then ideally results in the query sequence amongst the highest hits; an orthologue is identified if a high-ranking hit in the first BLAST is not from the same species as from which the query sequence is derived, and preferably results upon BLAST back in the query sequence being among the highest hits.

High-ranking hits are those having a low E-value. The lower the E-value, the more significant the score (or in other words the lower the chance that the hit was found by chance). Computation of the E-value is well known in the art. In addition to E-values, comparisons are also scored by percentage identity. Percentage identity refers to the number of identical nucleotides (or amino acids) between the two compared nucleic acid (or polypeptide) sequences over a particular length. In the case of large families, ClustalW may be used, followed by a neighbour joining tree, to help visualize clustering of related genes and to identify orthologues and paralogues.

The term " sequence identity" between two nucleic acid sequences is understood as meaning the percent identity of the nucleic acid sequence over in each case the entire sequence length which is calculated by alignment with the aid of the program algorithm GAP (Wisconsin Package Version 10.0, University of Wisconsin, Genetics Computer Group (GCG), Madison, USA), setting the following parameters:

Gap Weight: 12 Length Weight: 4

Average Match: 2,912 Average Mismatch:-2,003

The term " sequence identity" between two amino acid sequences is understood as meaning the percent identity of the nucleic acid sequence over in each case the entire sequence length which is calculated by alignment with the aid of the program algorithm GAP (Wisconsin Package Version 10.0, University of Wisconsin, Genetics Computer Group (GCG), Madison, USA), setting the following parameters:

Gap Weight: 8 Length Weight: 2

Average Match: 2,912 Average Mismatch:-2,003

The term "hybridisation" as defined herein is a process wherein substantially homologous complementary nucleotide sequences anneal to each other. The hybridisation process can occur entirely in solution, i.e. both complementary nucleic acids are in solution. The hybridisation process can also occur with one of the complementary nucleic acids immobilised to a matrix such as magnetic beads, Sepharose beads or any other resin. The hybridisation process can furthermore occur with one of the complementary nucleic acids immobilised to a solid support such as a nitro-cellulose or nylon membrane or immobilised by e.g. photolithography to, for example, a siliceous glass support (the latter known as nucleic acid arrays or microarrays or as nucleic acid chips). In order to allow hybridisation to occur, the nucleic acid molecules are generally thermally or chemically denatured to melt a double strand into two single strands and/or to remove hairpins or other secondary structures from single stranded nucleic acids.

The term " stringency" refers to the conditions under which a hybridisation takes place. The stringency of hybridisation is influenced by conditions such as temperature, salt concentration, ionic strength and hybridisation buffer composition. Generally, low stringency conditions are selected to be about 30°C lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. Medium stringency conditions are when the temperature is 20°C below T _m , and high stringency conditions are when the temperature is 10°C below T _m . High stringency hybridisation conditions are typically used for isolating hybridising sequences that have high sequence similarity to the target nucleic acid sequence. However, nucleic acids may deviate in sequence and still encode a substantially identical polypeptide, due to the degeneracy of the genetic code. Therefore medium stringency hybridisation conditions may sometimes be needed to identify such nucleic acid molecules.

The Tm is the temperature under defined ionic strength and pH, at which 50% of the target sequence hybridises to a perfectly matched probe. The T _m is dependent upon the solution conditions and the base composition and length of the probe. For example, longer sequences hybridise specifically at higher temperatures. The maximum rate of hybridisation is obtained from about 16°C up to 32°C below T _m . The presence of monovalent cations in the hybridisation solution reduce the electrostatic repulsion between the two nucleic acid strands thereby promoting hybrid formation; this effect is visible for sodium concentrations of up to 0.4M (for higher concentrations, this effect may be ignored). Formamide reduces the melting temperature of DNA-DNA and DNA-RNA duplexes with 0.6 to 0.7°C for each percent formamide, and addition of 50% formamide allows hybridisation to be performed at 30 to 45°C, though the rate of hybridisation will be lowered. Base pair mismatches reduce the hybridisation rate and the thermal stability of the duplexes. On average and for large probes, the Tm decreases about 1 °C per % base mismatch. The T _m may be calculated using the following equations, depending on the types of hybrids:

1 ) DNA-DNA hybrids (Meinkoth and Wahl, Anal. Biochem., 138: 267-284, 1984):

T _m = 81.5°C + 16.6xlogi ₀ [Na ⁺ ] ^a + 0.41x%[G/C ^b ] - 500x[L ^c ]- ¹ - 0.61x% formamide

2) DNA-RNA or RNA-RNA hybrids:

T _m = 79.8°C+ 18.5 (logi ₀ [Na ⁺ ] ^a ) + 0.58 (%G/C ^b ) + 1 1.8 (%G/C ^b ) ² - 820/L ^c

3) oligo-DNA or oligo-RNA ^d hybrids:

For <20 nucleotides: T _m = 2 (l _n )

For 20- 35 nucleotides: T _m = 22 + 1.46 (l _n )

a or for other monovalent cation, but only accurate in the 0.01- 0.4 M range.

b only accurate for %GC in the 30% to 75% range.

c L = length of duplex in base pairs.

d oligo, oligonucleotide; l _n , = effective length of primer = 2χ(ηο. of G/C)+(no. of A T). Non-specific binding may be controlled using any one of a number of known techniques such as, for example, blocking the membrane with protein containing solutions, additions of heterologous RNA, DNA, and SDS to the hybridisation buffer, and treatment with Rnase. For non-homologous probes, a series of hybridizations may be performed by varying one of (i) progressively lowering the annealing temperature (for example from 68°C to 42°C) or (ii) progressively lowering the formamide concentration (for example from 50% to 0%). The skilled artisan is aware of various parameters which may be altered during hybridisation and which will either maintain or change the stringency conditions.

Besides the hybridisation conditions, specificity of hybridisation typically also depends on the function of post-hybridisation washes. To remove background resulting from non- specific hybridisation, samples are washed with dilute salt solutions. Critical factors of such washes include the ionic strength and temperature of the final wash solution: the lower the salt concentration and the higher the wash temperature, the higher the stringency of the wash. Wash conditions are typically performed at or below hybridisation stringency. A positive hybridisation gives a signal that is at least twice of that of the background.

Generally, suitable stringent conditions for nucleic acid hybridisation assays or gene amplification detection procedures are as set forth above. More or less stringent conditions may also be selected. The skilled artisan is aware of various parameters which may be altered during washing and which will either maintain or change the stringency conditions. For example, typical high stringency hybridisation conditions for DNA hybrids longer than 50 nucleotides encompass hybridisation at 65°C in 1x SSC or at 42°C in 1x SSC and 50% formamide, followed by washing at 65°C in 0.3x SSC. Examples of medium stringency hybridisation conditions for DNA hybrids longer than 50 nucleotides encompass

hybridisation at 50°C in 4x SSC or at 40°C in 6x SSC and 50% formamide, followed by washing at 50°C in 2x SSC. The length of the hybrid is the anticipated length for the hybridising nucleic acid. When nucleic acids of known sequence are hybridised, the hybrid length may be determined by aligning the sequences and identifying the conserved regions described herein. 1 xSSC is 0.15M NaCI and 15mM sodium citrate; the hybridisation solution and wash solutions may additionally include 5x Denhardt's reagent, 0.5-1.0% SDS, 100 μ g/ml denatured, fragmented salmon sperm DNA, 0.5% sodium pyrophosphate.

For the purposes of defining the level of stringency, reference can be made to Sambrook et al. (2001) Molecular Cloning: a laboratory manual, 3 ^rd Edition, Cold Spring Harbor

Laboratory Press, CSH, New York or to Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989 and yearly updates).

" Homologues" of a protein encompass peptides, oligopeptides, polypeptides, proteins and enzymes having amino acid substitutions, deletions and/or insertions relative to the unmodified protein in question and having similar biological and functional activity as the unmodified protein from which they are derived.

A deletion refers to removal of one or more amino acids from a protein. An insertion refers to one or more amino acid residues being introduced into a

predetermined site in a protein. Insertions may comprise N-terminal and/or C-terminal fusions as well as intra-sequence insertions of single or multiple amino acids. Generally, insertions within the amino acid sequence will be smaller than N- or C-terminal fusions, of the order of about 1 to 10 residues. Examples of N- or C-terminal fusion proteins or peptides include the binding domain or activation domain of a transcriptional activator as used in the yeast two-hybrid system, phage coat proteins, (histidine)-6-tag, glutathione S- transferase-tag, protein A, maltose-binding protein, dihydrofolate reductase, Tag* 100 epitope, c-myc epitope, FLAG ^® -epitope, lacZ, CMP (calmodulin-binding peptide), HA epitope, protein C epitope and VSV epitope.

A substitution refers to replacement of amino acids of the protein with other amino acids having similar properties (such as similar hydrophobicity, hydrophilicity, antigenicity, propensity to form or break a -helical structures or β -sheet structures). Amino acid substitutions are typically of single residues, but may be clustered depending upon functional constraints placed upon the polypeptide and may range from 1 to 10 amino acids; insertions will usually be of the order of about 1 to 10 amino acid residues. The amino acid substitutions are preferably conservative amino acid substitutions. Conservative substitution tables are well known in the art (see for example Creighton (1984) Proteins. W.H. Freeman and Company (Eds) and Table 1 below).

Table 2: Examples of conservative amino acid substitutions

Reference herein to an " endogenous" gene not only refers to the gene in question as found in an organism in its natural form (i.e., without there being any human intervention), but also refers to that same gene (or a substantially homologous nucleic acid/gene) in an isolated form subsequently (re)introduced into a microorganism (a transgene). For example, a transgenic microorganism containing such a transgene may encounter a substantial reduction of the transgene expression and/or substantial reduction of expression of the endogenous gene. The isolated gene may be isolated from an organism or may be manmade, for example by chemical synthesis. The terms " orthologues" and " paralogues" encompass evolutionary concepts used to describe the ancestral relationships of genes. Paralogues are genes within the same species that have originated through duplication of an ancestral gene; orthologues are genes from different organisms that have originated through speciation, and are also derived from a common ancestral gene.

The term " splice variant" as used herein encompasses variants of a nucleic acid sequence in which selected introns and/or exons have been excised, replaced, displaced or added, or in which introns have been shortened or lengthened. Such variants will be ones in which the biological activity of the protein is substantially retained; this may be achieved by selectively retaining functional segments of the protein. Such splice variants may be found in nature or may be manmade. Methods for predicting and isolating such splice variants are well known in the art (see for example Foissac and Schiex (2005) BMC Bioinformatics 6: 25).

The term " vector" , preferably, encompasses phage, plasmid, fosmid, cosmid, viral vectors as well as artificial chromosomes, such as bacterial or yeast artificial chromosomes. Moreover, the term also relates to targeting constructs which allow for random or site- directed integration of the targeting construct into genomic DNA. Such target constructs, preferably, comprise DNA of sufficient length for either homologous or heterologous recombination as described in detail below. The vector encompassing the polynucleotide of the present invention, preferably, further comprises selectable markers for propagation and/or selection in a recombinant microorganism. The vector may be incorporated into a recombinant microorganism by various techniques well known in the art. If introduced into a recombinant microorganism, the vector may reside in the cytoplasm or may be incorporated into the genome. In the latter case, it is to be understood that the vector may further comprise nucleic acid sequences which allow for homologous recombination or

heterologous insertion. Vectors can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques.

The terms " transformation" and " transfection" , conjugation and transduction, as used in the present context, are intended to comprise a multiplicity of prior-art processes for introducing foreign nucleic acid (for example DNA) into a recombinant microorganism, including calcium phosphate, rubidium chloride or calcium chloride co-precipitation, DEAE- dextran-mediated transfection, lipofection, natural competence, carbon-based clusters, chemically mediated transfer, electroporation or particle bombardment. Methods for many species of microorganisms are readily available in the literature, for example, in Turgeon (2010) Molecular and cell biology methods for fungi, p3-9, in Koushki, MM et al., (201 1 ), AFRICAN JOURNAL OF BIOTECHNOLOGY Vol.10 (41): p7939-7948, in Coyle et al.

(2010) Appl Environ Microbiol 76:3898- 3903, in Current Protocols in Molecular Biology, Chapter 13. Eds Ausubel F.M. et al. Wiley & Sons, U.K., and in Genome Analysis: A

Laboratory Manual, Cloning Systems. Volume 3. Edited by Birren B, Green ED, Klapholz S, Myers RM, Riethman H, Roskams J. New York: Cold Spring Harbor Laboratory Press; 1999:297-565; Kieser et al. (2000), Practical Streptomyces Genetics, John Innes

Foundation.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

In a first aspect, the present invention provides nucleic acid sequences which

encodespolypeptides being involved in pamamycin synthesis. As used herein, pamamycin synthesis encompasses all steps of the biosynthesis of pamamycin. Accordingly, a polypeptide which is involved in the synthesis of pamamycin may either convert a substrate into pamamycin or may produce any of the precursors which occur in pamamycin biosynthesis. Preferably, the polypeptide encoded by the polynucleotide of the present invention shall be capable of increasing the amount of pamamycin or a precursor thereof upon expression in an organism, preferably a recombinant microorganism as specified elsewhere herein. Precursors of pamamycin are depicted in Figure 2. Such an increase is, preferably, statistically significant when compared to a control organism which lacks expression of the polynucleotide of the present invention. Preferably the control organism is of the same species and even more preferred belongs to the same strain that was used to construct the recombinant microorganism. Whether an increase is significant can be determined by statistical tests well known in the art including, e.g., Student ^' s t-test. More preferably, the increase is an increase of the amount of pamamycin of at least 5%, at least 10%, at least 15%, at least 20% or at least 30% compared to said control. Suitable assays to identify and measure the amount of pamamycin are known by the person skilled in the art and are described for example in Hashimoto et al (2004) Biosynthetic Origin of the Carbon Skeleton and Nitrogen Atom of Pamamycin-607, a Nitrogen-Containing Polyketide, Biosci. Biotechnol. Biochem. 69; Hashimoto et al. (201 1 ) Effect of Pamamycin-607 on Secondary Metabolite Production by Streptomyces spp., Biosci. Biotechnol. Biochem. 75; Hashimoto et al. (2003) Relationship between Response to and Production of the Aerial Mycelium- inducing Substances Pamamycin-607 and A-factor, Biosci. Biotechnol. Biochem. 67;

Natsume et al. (1991 ) The Structures of Four New Pamamycin Homologues Isolated from Streptomyces alboniger, Tetrahedron Letters 32.

One nucleic acid sequence provided by the invention is a gene cluster described by SEQ ID NO: 1 , which comprises expression cassette for polypeptides being involved in pamamycin synthesis. A schematic representation of the gene cluster is provided by Figure 1 .

Table 3: Listing of genes and encoded polypeptides of the polnucleotidesequence described by SEQ ID NO: 2:

Gene Gene Starting Endpoint Seq Function Correspond in

Number Name point in in SEQ ID I D g protein Seq

(ORF) SEQ ID No No 1 NO: ID NO:

1

0726 pamR2 18 755 129 TetR type 130

regulator

0727 pamW 843 2519 136 transporter 137

0729 pamC 2616 2846 3 ACP 4

0730 pamG 2900 3934 10 KS 1 1

0731 pamF 3946 5265 17 KS 18

0732 pamA 5477 6814 24 KS 25

0733 pamB 681 1 8673 31 Acyl-CoA transf. 32

0734 pamD 871 1 9670 38 KS 39

0736 pamE 9723 10757 45 KS 46

0737 pamO 1081 1 1 1605 52 KR 53

0738 pamK 1 1621 12700 59 KS 60

0740 pamJ 12717 14012 66 KS 67

0741 pamM 14038 15582 73 KR 74 0742 pamN 15579 16406 80 DH/KR 81

0743 pamL 16403 18106 87 Acyl-CoA ligase 88

0744 pamD 18090 19073 122 hydrolase 123

0745 pamX 19170 20516 94 amino 95

transferase

0746 pamY 20565 21386 101 methyl 102

transferase

0747 pamS 21604 22410 108 enoyl-CoA 109

hydratase

0749 pamR1 22435 23082 1 15 LuxR Transcrf. 1 16

Table 3 provides a listing of the polypeptide (protein) encoding sequences of SEQ ID NO: 2, the respective ORF names, the number of the nucleotides in SEQ ID NO: 2, which are starting and endpoints of the polypeptide encoding sequences, the likely function of the encoded polypeptides and the respective SEQ ID NOs: of the polynucleotide and amino acid sequences in the sequence listings. The starting points and end points given in Table 3 do not necessarily represent the 5-prime and 3-prime ends of the polypeptide encoding regions. The orientation of the transcribed region is depicted in the sketch of Cluster 5 in Figure 1.

The provided recombinant polynucleotides can either be isolated from their natural genomic environment and modified after their isolation or produced artificially from pure sequence information. A natural source of polynucleotides of the invention are pamamycin producing organisms and related species. One way to isolate polynucleotides from pamamycin producing organisms is to perform a PCR reaction on genomic DNA of such organisms using primers comprising at least 15, preferably at least 20, consecutive polynucleotides of the 5-prime and 3-prime ends of the polypeptide encoding sequences disclosed in SEQ ID NO: 1 and/or 2. Pamamycin producing organisms can, for example but not excluding others, be found in the group consisting of the genus Streptomyces preferably of the species Streptomyces alboniger, Streptomyces aurantiacus, or Streptomyces kitasatoensis, in particular in the strains Streptomyces alboniger ATCC 12461 Streptomyces alboniger DSM 40043, Streptomyces aurantiacus ATCC 19822 and Streptomyces kitasatoensis JCM5001 ,. Pamamycin producing organisms can also be used to isolate or identify variants of SEQ ID NO: 1 and/or 2, by using PCR and/or other hybridisation techniques and/or by using bioinformatic searches of available genomic sequence information. Variants of SEQ ID NO: 1 and/or 2 have preferably polypeptide encoding regions for most, preferably all of the polypeptides of the group of polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and/or pamS activity.

Strains of these species are deposited, for example, as Streptomyces alboniger ATCC 12461 also deposited as DSM 40043 and IFO 12738 and isolated in 1952 from forest soil in Wisconsin (USA), Streptomyces aurantiacus ATCC 19822 also deposited as DSM 40412, Streptomyces aurantiacus I MET 43917, Streptomyces aurantiacus J A 4570 and

Streptomyces kitasatoensis JCM5001. These strains are deposited under the respective deposit numbers and disclosed in DE4134168, DE4316836, US4283391 , JP62135476A, in Hashimoto et al. in Biosci. Biotechnol. Biochm. 2003, Vol 67(4), pages 803 to 808 and/or HAERTL et al. THE JOURNAL OF ANTIBIOTICS (1998) VOL. 51 , NO. 1 1 , pp. 1040-1046.

The polynucleotides and polypeptides provided by the present invention are suitbable to identify and construct recombinant microorganisms having the ability to produce pamamycin or to produce recombinant versions of pamamycin producing microorganisms which have an enhanced or modified production of pamamycin.

Accordingly, the invention includes microorganisms comprising:

a) at least one expression cassette encoding a polypeptide having pamC activity, and b) at least one expression cassette encoding a polypeptide having pamG activity, and c) at least one expression cassette encoding a polypeptide having pamF activity, and d) at least one expression cassette encoding a polypeptide having pamA activity, and e) at least one expression cassette encoding a polypeptide having pamB activity, and f) at least one expression cassette encoding a polypeptide having pamD activity, and g) at least one expression cassette encoding a polypeptide having pamE activity, and h) at least one expression cassette encoding a polypeptide having pamO activity, and i) at least one expression cassette encoding a polypeptide having pamK activity, and j) at least one expression cassette encoding a polypeptide having pamJ activity, and k) at least one expression cassette encoding a polypeptide having pamM activity, and

I) at least one expression cassette encoding a polypeptide having pamN activity, and m) at least one expression cassette encoding a polypeptide having pamL activity, and n) at least one expression cassette encoding a polypeptide having pamX activity, and o) at least one expression cassette encoding a polypeptide having pamY activity, and

P) at least one expression cassette encoding a polypeptide having pamS activity.

Microorganisms comprising at least one expression cassette for the polypeptide activities a) to p) can be identified by testing microorganisms for the production of pamamycin and/or for the presence of polypeptide encoding regions for at least 3 activities selected from pamC activity, pamG activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity and pamS activity. Microorganism which are selected to be tested for the production of pamamycin and/or to be used as sources to obtain sequence variants of SEQ ID NO: 1 or 2 or at least one of the polpypetide encoding regions, promoters or terminators comprised therein, have in order of preference

polypeptide encoding regions for at least 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, or all of the activities mentiond above. Also preferred is to test the microorganism for the production of pamamycin, preferably for the production of at least one of pamamycin 607, pamamycin 621 A, pamamycin 621 B, pamamycin 621C or pamamycin 621 D.

Microorganisms can be tested for the presence of expression cassettes for each one of the polypeptide activities of a) to p), by several ways known to the person skilled in the art. For example, but not excluding others, by testing the genome of a microorganism for

sequences hybridizing under stringent conditions to polynucleotide probes covering at least 50, 100, 200, 300, 400, 500, 600 or at least 700 consecutive polynucleotides of at least one of the polypeptide encoding regions for pamC activity, pamG activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity and pamS activity comprised by SEQ I D NO: 1.

Another way to test for expression cassettes for each one of the polypeptide activities of a) to p) is to perform sequence comparisons of sequenced DNA of the microorganism, in order to identify DNA sequences having at least 70%, 80% or 90% sequence identity to at least one of the polypeptide encoding regions for pamC activity, pam G activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity and pamS activity comprised by SEQ ID NO: 1.

A further way to test for expression cassettes for each one of the polypeptide activities of a) to p) in a given microorganisms is to perform sequence comparisons of sequenced RNA of the microorganism in question, in order to identify microorganisms comprising mRNAs being at least 70%, 80% or 90% identical to at least one of the polypeptide encoding regions for pamC activity, pamG activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity and pamS activity comprised by SEQ ID NO: 1.

The recombinant microorgaisms of the invention are preferably microorganisms which have been intentionally modified via molecular biology techniques, like transformation or gene knock-out or gene replacement. However, this term recombinant microorganism does also apply to non-naturally occurring microorganisms. Non-naturally occurring microorganisms are microorganisms which have been created by single or repeated mutagenesis and selection performed on microorganism isolated from nature. Non-naturally occurring microorganisms comprise at least one up-regulated activity of a polypeptide having pamC activity, pamG activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity, pamS activity, pamR1 activity, pamW activity, and/or at least one down-regulated pamR2 activity or pamH activity, in comparison to the unmodified version of such microorganism. Recombinant microorganisms are preferably produced via transformation of a

microorganism with polynucletide constructs comprising at least one expression cassette encoding a polypeptide having pamC activity, pamG activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity, pamS activity, pamR1 activity, pamW activity, pamR2 activity or pamD activity. Microorganisms can also be transformed with polynucleotide constructs which are able to modify expression of at least one expression cassette encoding such an activity and which is already present in such microorganism. Such polynucleotide construes may replace the endogenous promoter with a heterologous promoter sequence providing for constitutive or inducible expression in such microorganism, or which provide for a stronger or weaker expression in such microorganism in comparison to the endogenous promoter. Alternatively, or in addition thereto, the expression of at least one expression cassette can also be modified by providing the microorganism with an expression cassette for a transcriptional regulator which enhances or decreases expression from such expression cassette, or to provide the organism with an expression cassette encoding a polypeptide having one of the above described activities, but providing such activity with a higher activity per expressed polypeptide molecule or having a higher stability of such polypeptide molecules in order to prolong enzymatic activity per expressed polypeptide.

Polypeptides having pamC activity:

A polypeptide having pamC activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 4, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 3, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 4. Preferably it is the same promoter.

The invention provides for polypeptides having pamC activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a complement of SEQ ID NO: 3.

Preferably the polynucleotides encoding polypeptides having pamC activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 3 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 3, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 3.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 4 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 4. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 4 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 5, 6, 7, 8 and 9. Further variants can be deduced by the information provided by Figure 3 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 4 shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 4, 5, 6, 7, 8 and 9. The parameters used for the pairwise sequence alignments are:

Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 4:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 4 NO: 5 NO: 6 NO: 7 NO: 8 NO: 9

SEQ ID NO: 4 100% 95% 95% 92% 89% 89%

SEQ ID NO: 5 - 100% 92% 89% 89% 87%

SEQ ID NO: 6 - - 100% 89% 88% 87%

SEQ ID NO: 7 - - - 100% 92% 84%

SEQ ID NO: 8 - - - - 100% 80%

SEQ ID NO: 9 - - - - - 100%

Further examples for polypeptides having pamC activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamC activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 209, and/or are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 208, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 208.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 208 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 208, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 208.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 209 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 209. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 209 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 210, 21 1 , 212, 213 and 214. Further variants can be deduced aligning the sequences of SEQ ID NO: 209, 210, 21 1 , 212, 213 and 214 to the information provided by Figure 3 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 4a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs 4, 209, 210, 21 1 , 212, 213 and 214. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 4a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 4 NO: 209 NO: 210 NO: 21 1 NO: 212 NO: 213 NO: 214

SEQ ID NO: 4 100% 86% 82% 80% 80% 76% 75%

SEQ ID NO: 209 - 100% 95% 95% 92% 89% 89%

SEQ ID NO: 210 - - 100% 95% 89% 89% 89%

SEQ ID NO: 21 1 - - - 100% 87% 87% 87%

SEQ ID NO: 212 - - - - 100% 84% 88%

SEQ ID NO: 213 - - - - - 100% 82%

SEQ ID NO: 214 - - - - - - 100%

Polypeptides having pamG activity:

A polypeptide having pamG activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 1 1 , or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 10, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ I D NO: 1 1. Preferably it is the same promoter.

The invention provides for polypeptides having pamC activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 1 1 , and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 10, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 10.

Preferably the polynucleotides encoding polypeptides having pamG activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 10 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 10, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 10.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 1 1 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%,

98%, 99%, or 100% identical SEQ ID NO: 1 1. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 1 1 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 12, 13, 14, 15 and 16. Further variants can be deduced by the information provided by Figure 4 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 5 shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 1 1 , 12, 13, 14, 15 and 16. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 5:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 1 1 NO: 12 NO: 13 NO: 14 NO: 15 NO: 16

SEQ ID NO 1 1 100% 95% 95% 93% 90% 90%

SEQ ID NO 12 - 100% 91 % 90% 88% 86%

SEQ ID NO 13 - - 100% 89% 88% 87%

SEQ ID NO 14 - - - 100% 88% 88%

SEQ ID NO 15 - - - - 100% 85%

SEQ ID NO 16 - - - - - 100%

Further examples for polypeptides having pamG activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamG activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 216, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 215, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 215.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 215 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 215, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 215.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 209 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 216. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 216 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 217, 218, 219, 220 and 221. Further variants can be deduced aligning the sequences of SEQ ID NO: 216, 217, 218, 219, 220 and 221 to the information provided by Figure 4 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 5a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 1 1 , 216, 217, 218, 219, 220 and 221 . The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 5a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 1 1 NO: 216 NO: 217 NO: 218 NO: 219 NO: 220 NO: 221

SEQ ID NO: 1 1 100% 90% 86% 85% 83% 81 % 81 %

SEQ ID NO: 216 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 217 - - 100% 90% 89% 87% 89%

SEQ ID NO: 218 - - - 100% 89% 88% 86%

SEQ ID NO: 219 - - - - 100% 85% 86%

SEQ ID NO: 220 - - - - - 100% 86%

SEQ ID NO: 221 - - - - - - 100%

Polypeptides having pamF activity:

A polypeptide having pamF activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 18, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 17, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 18. Preferably it is the same promoter.

The invention provides for polypeptides having pamC activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 17. Preferably the polynucleotides encoding polypeptides having pamF activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 17 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 17, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 17.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 18 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 18. These sequence variants can be obtained by substitution of one or more amino acids of SEQ I D NO: 18 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 19, 20, 21 , 22 and 23. Further variants can be deduced by the information provided by Figure 5 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 6 shows the % sequence identity of pairwise sequence alignments of SEQ I D NO: 18, 19, 20, 21 , 22 and 23. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 6:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 18 NO: 19 NO: 20 NO: 21 NO: 22 NO: 23

SEQ ID NO: 18 100% 95% 95% 93% 90% 90%

SEQ ID NO: 19 - 100% 91 % 89% 87% 89%

SEQ ID NO: 20 - - 100% 89% 87% 87%

SEQ ID NO: 21 - - - 100% 87% 87%

SEQ ID NO: 22 - - - - 100% 83%

SEQ ID NO: 23 - - - - - 100% Further examples for polypeptides having pamF activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamF activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 223, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 222, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 222.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 222 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 222, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 222.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 223 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 223. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 223 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 224, 225, 226, 227 and 228. Further variants can be deduced aligning the sequences of SEQ ID NO: 223, 224, 225, 226, 227 and 228 to the information provided by Figure 5 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 6a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 18, 223, 224, 225, 226, 227 and 228. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 6a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 18 NO: 223 NO: 224 NO: 225 NO: 226 NO: 227 NO: 228

SEQ ID NO: 18 100% 92% 92% 90% 86% 84% 83%

SEQ ID NO: 223 - 100% 100% 95% 93% 90% 90%

SEQ ID NO: 224 - - 100% 95% 93% 90% 90%

SEQ ID NO: 225 - - - 100% 90% 90% 86%

SEQ ID NO: 226 - - - - 100% 88% 85%

SEQ ID NO: 227 - - - - - 100% 85%

SEQ ID NO: 228 - - - - - - 100% Polypeptides having pamA activity:

A polypeptide having pamA activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 25, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 24, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 25. Preferably it is the same promoter.

The invention provides for polypeptides having pamA activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 24, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 24.

Preferably the polynucleotides encoding polypeptides having pamA activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 24 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 24, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 24.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 25 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 25. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 25 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 26, 27, 28, 29 and 30. Further variants can be deduced by the information provided by Figure 6 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 7 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 25, 26, 27, 28, 29 and 30. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 7:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 25 NO: 26 NO: 27 NO: 28 NO: 29 NO: 30

SEQ ID NO: 25 100% 95% 95% 93% 90% 90%

SEQ ID NO: 26 - 100% 91 % 89% 87% 87%

SEQ ID NO: 27 - - 100% 90% 87% 88%

SEQ ID NO: 28 - - - 100% 87% 86%

SEQ ID NO: 29 - - - - 100% 84%

SEQ ID NO: 30 - - - - - 100%

Further examples for polypeptides having pamA activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamA activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 230, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 229, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 229.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 229 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 229, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 229.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 230 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 230. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 230 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 231 , 232, 233, 234 and 235. Further variants can be deduced aligning the sequences of SEQ ID NO: 230, 231 , 232, 233, 234 and 235 to the information provided by Figure 6 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 7a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 25, 230, 231 , 232, 233, 234 and 235. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 7a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 25 NO: 230 NO: 231 NO: 232 NO: 233 NO: 234 NO: 235

SEQ ID NO: 25 100% 92% 88% 88% 86% 84% 83%

SEQ ID NO: 230 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 231 - - 100% 100% 89% 88% 87%

SEQ ID NO: 232 - - - 100% 89% 88% 87%

SEQ ID NO: 233 - - - - 100% 86% 86%

SEQ ID NO: 234 - - - - - 100% 82%

SEQ ID NO: 235 - - - - - - 100%

Polypeptides having pamB activity:

A polypeptide having pamB activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 32, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 31 , in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 32. Preferably it is the same promoter.

The invention provides for polypeptides having pamB activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31 , or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 31.

Preferably the polynucleotides encoding polypeptides having pamB activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 31 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 31 , using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 31.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 32 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 32. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 32 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 33, 34, 35, 36 and 37. Further variants can be deduced by the information provided by Figure 7 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 8 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 32, 33, 34, 35, 36 and 37. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 8:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 32 NO: 33 NO: 34 NO: 35 NO: 36 NO: 37

SEQ ID NO: 32 100% 95% 95% 93% 90% 90%

SEQ ID NO: 33 - 100% 92% 90% 88% 89%

SEQ ID NO: 34 - - 100% 90% 88% 87%

SEQ ID NO: 35 - - - 100% 87% 88%

SEQ ID NO: 36 - - - - 100% 87%

SEQ ID NO: 37 - - - - - 100%

Further examples for polypeptides having pamB activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamB activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 237, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 236, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a complement of SEQ ID NO: 236.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 236 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 236, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 236.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 237 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 237. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 237 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ I D NO: 238, 239, 240, 241 and 242. Further variants can be deduced aligning the sequences of SEQ ID NO: 237, 238, 239, 240, 241 and 242 to the information provided by Figure 7 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 8a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 32, 237, 238, 239, 240, 241 and 242. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62. Table 8a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 32 NO: 237 NO: 238 NO: 239 NO: 240 NO: 241 NO: 242

SEQ ID NO: 32 100% 88% 84% 84% 82% 79% 80%

SEQ ID NO: 237 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 238 - - 100% 92% 90% 88% 89%

SEQ ID NO: 239 - - - 100% 91 % 89% 88%

SEQ ID NO: 240 - - - - 100% 87% 88%

SEQ ID NO: 241 - - - - - 100% 86%

SEQ ID NO: 242 - - - - - - 100%

Polypeptides having pamB activity which are derived from other metabolic pathways

Further examples for polypeptides having pamB activity are polypeptides being obtainable from Streptomyces fulvissimus and their variants.

Thus, the invention provides for polypeptides having pamB activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 349, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 348, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a complement of SEQ ID NO: 348.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 348 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 348, using genomic DNA or cDNA of Streptomyces fulvissimus as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 348. The invention does also include sequence variants of polypeptides of SEQ ID NO: 349 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 349. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 349 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 350, 351 , 352, 353 and 354. Further variants can be deduced aligning the sequences of SEQ ID NO: 349, 350, 351 , 352, 353 and 354 to the information provided by Figure 23 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 8b shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 349, 350, 351 , 352, 353 and 354. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 8b:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 349 NO: 350 NO: 351 NO: 352 NO: 353 NO: 354

SEQ ID NO: 349 100% 95% 95% 93% 90% 90%

SEQ ID NO: 350 - 100% 92% 89% 89% 87%

SEQ ID NO: 351 - - 100% 91 % 88% 89%

SEQ ID NO: 352 - - - 100% 88% 87%

SEQ ID NO: 353 - - - - 100% 87%

SEQ ID NO: 354 - - - - - 100%

Further examples for polypeptides having pamB activity are polypeptides being obtainable from Streptomyces griseus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamB activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 356, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 355, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a complement of SEQ ID NO: 355.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 355 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 355, using genomic DNA or cDNA of Streptomyces griseus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 355. The invention does also include sequence variants of polypeptides of SEQ ID NO: 356 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 356. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 356 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 357, 358, 359, 360 and 361. Further variants can be deduced aligning the Sequences of SEQ ID NO: 356, 357, 358, 359, 360 and 361 to the information provided by Figure 23 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 8c shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 356, 357, 358, 359, 360 and 361 . The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 8c:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 356 NO: 357 NO: 358 NO: 359 NO: 360 NO: 361

SEQ ID NO: 356 100% 95% 95% 93% 90% 90%

SEQ ID NO: 357 - 100% 92% 90% 88% 90%

SEQ ID NO: 358 - - 100% 91 % 88% 87%

SEQ ID NO: 359 - - - 100% 87% 86%

SEQ ID NO: 360 - - - - 100% 88%

SEQ ID NO: 361 - - - - - 100%

Further examples for polypeptides having pamB activity are polypeptides being obtainable from Streptomyces sp. SolWspMP and their variants.

Thus, the invention provides for polypeptides having pamB activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 363, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 362, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a complement of SEQ ID NO: 362.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 362 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 362, using genomic DNA or cDNA of Streptomyces sp. SolWspMP as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 362. The invention does also include sequence variants of polypeptides of SEQ ID NO: 363 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 363. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 363 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 364, 365, 366, 367 and 368. Further variants can be deduced aligning the sequences of SEQ ID NO: 363, 364, 365, 366, 367 and 368 to the information provided by Figure 23 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 8d shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 363, 364, 365, 366, 367 and 368. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 8d:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 363 NO: 364 NO: 365 NO: 366 NO: 367 NO: 368

SEQ ID NO: 363 100% 95% 95% 93% 90% 90%

SEQ ID NO: 364 - 100% 91 % 90% 89% 87%

SEQ ID NO: 365 - - 100% 89% 90% 88%

SEQ ID NO: 366 - - - 100% 87% 86%

SEQ ID NO: 367 - - - - 100% 86%

SEQ ID NO: 368 - - - - - 100%

Further examples for polypeptides having pamB activity are polypeptides being obtainable from Streptomyces sp. W007 and their variants.

Thus, the invention provides for polypeptides having pamB activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 370, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 369, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a complement of SEQ ID NO: 369.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 369 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 369, using genomic DNA or cDNA of Streptomyces sp. W007 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 369. The invention does also include sequence variants of polypeptides of SEQ ID NO: 370 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 370. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 370 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 371 , 372, 373, 374 and 375. Further variants can be deduced aligning the Sequences of SEQ ID NO: 370, 371 , 372, 373, 374 and 375 to the information provided by Figure 23 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 8e shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 370, 371 , 372, 373, 374 and 375. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 8e:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 370 NO: 371 NO: 372 NO: 373 NO: 374 NO: 375

SEQ ID NO: 370 100% 95% 95% 95% 90% 90%

SEQ ID NO: 371 - 100% 91 % 91 % 89% 88%

SEQ ID NO: 372 - - 100% 100% 87% 88%

SEQ ID NO: 373 - - - 100% 87% 88%

SEQ ID NO: 374 - - - - 100% 85%

SEQ ID NO: 375 - - - - - 100%

Further examples for polypeptides having pamB activity are polypeptides being obtainable from Streptomyces californicus and their variants.

Thus, the invention provides for polypeptides having pamB activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 377, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 376, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 376.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 376 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 376, using genomic DNA or cDNA of Streptomyces californicus as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 376. The invention does also include sequence variants of polypeptides of SEQ ID NO: 377 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 377. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 377 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 378, 379, 380, 381 and 382. Further variants can be deduced aligning the sequences of SEQ ID NO: 377, 378, 379, 380, 381 and 382 to the information provided by Figure 23 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 8f shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 377, 378, 379, 380, 381 and 382. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 8f:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 377 NO: 378 NO: 379 NO: 380 NO: 381 NO: 382

SEQ ID NO: 377 100% 95% 95% 93% 90% 90%

SEQ ID NO: 378 - 100% 91 % 90% 87% 88%

SEQ ID NO: 379 - - 100% 90% 88% 89%

SEQ ID NO: 380 - - - 100% 87% 88%

SEQ ID NO: 381 - - - - 100% 85%

SEQ ID NO: 382 - - - - - 100%

Table 8g shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs 32, 237, 349, 356, 363, 370 and 377. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 8g:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 32 NO: 237 NO: 349 NO: 356 NO: 363 NO: 370 NO: 377

SEQ ID NO: 32 100% 88% 58% 59% 57% 59% 59%

SEQ ID NO: 237 - 100% 57% 58% 58% 58% 57% SEQ ID NO: 349 100% 90% 92% 90% 90%

SEQ ID NO: 356 - 100% 88% 97% 90%

SEQ ID NO: 363 - - 100% 88% 88%

SEQ ID NO: 370 - - - 100% 89%

SEQ ID NO: 377 - - - - 100%

Polypeptides having pamD activity:

A polypeptide having pamD activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 39, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 38, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 39. Preferably it is the same promoter. The invention provides for polypeptides having pamD activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 38.

Preferably the polynucleotides encoding polypeptides having pamD activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 38 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 38, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 38. Further variants can be deduced by the information provided by Figure 8 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. The invention does also include sequence variants of polypeptides of SEQ ID NO: 39 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 39. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 39 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 40, 41 , 42, 43 and 44.

Table 9 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 39, 40, 41 , 42, 43 and 44. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 9:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 39 NO: 40 NO: 41 NO: 42 NO: 43 NO: 44

SEQ ID NO: 39 100% 95% 95% 93% 90% 90%

SEQ ID NO: 40 - 100% 91 % 90% 89% 87%

SEQ ID NO: 41 - - 100% 90% 87% 85%

SEQ ID NO: 42 - - - 100% 85% 86%

SEQ ID NO: 43 - - - - 100% 86%

SEQ ID NO: 44 - - - - - 100% Further examples for polypeptides having pamD activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamD activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 244, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 243, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 243.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 243 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 243, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 243. The invention does also include sequence variants of polypeptides of SEQ ID NO: 244 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 244. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 244 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ I D NO: 245, 246, 247, 248 and 249. Further variants can be deduced aligning the sequences of SEQ ID NO: 244, 245, 246, 247, 248 and 249 to the information provided by Figure 8 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 9a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 39, 244, 245, 246, 247, 248 and 249. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 9a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 39 NO: 244 NO: 245 NO: 246 NO: 247 NO: 248 NO: 249

SEQ ID NO: 39 100% 93% 88% 89% 86% 83% 84%

SEQ ID NO: 244 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 245 - - 100% 91 % 90% 88% 87%

SEQ ID NO: 246 - - - 100% 90% 87% 86%

SEQ ID NO: 247 - - - - 100% 88% 85%

SEQ ID NO: 248 - - - - - 100% 85%

SEQ ID NO: 249 - - - - - - 100% Polypeptides having pamE activity:

A polypeptide having pamE activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 46, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 45, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 46. Preferably it is the same promoter. The invention provides for polypeptides having pamE activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 45.

Preferably the polynucleotides encoding polypeptides having pamE activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 45 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 45, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 45.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 46 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 46. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 46 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 47, 48, 49, 50 and 51. Further variants can be deduced by the information provided by Figure 9 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 10 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 46, 47, 48, 49, 50 and 51. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 10:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 46 NO: 47 NO: 48 NO: 49 NO: 50 NO: 51

SEQ ID NO: 46 100% 95% 95% 93% 90% 90%

SEQ ID NO: 47 - 100% 90% 90% 86% 86%

SEQ ID NO: 48 - - 100% 89% 88% 86%

SEQ ID NO: 49 - - - 100% 85% 85%

Further examples for polypeptides having pamE activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants. Thus, the invention provides for polypeptides having pamE activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 251 , and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 250, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 250.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 250 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 250, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 250. The invention does also include sequence variants of polypeptides of SEQ ID NO: 251 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 251. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 251 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 252, 253, 254, 255 and 256. Further variants can be deduced aligning the sequences of SEQ ID NO: 251 , 252, 253, 254, 255 and 256 to the information provided by Figure 9 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 10a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 46, 251 , 252, 253, 254, 255 and 256. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 10a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 46 NO: 251 NO: 252 NO: 253 NO: 254 NO: 255 NO: 256

SEQ ID NO: 46 100% 94% 89% 89% 88% 84% 84% SEQ ID NO: 251 100% 95% 95% 93% 90% 90% SEQ ID NO: 252 100% 90% 90% 88% 85% SEQ ID NO: 253 100% 90% 90% 90% SEQ ID NO: 254 100% 85% 84%

SEQ ID NO: 255 - 100% 85%

SEQ ID NO: 256 - - 100%

Polypeptides having pamO activity:

A polypeptide having pamO activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 53, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 52, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 53. Preferably it is the same promoter. The invention provides for polypeptides having pamO activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 54, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 52, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 52.

Preferably the polynucleotides encoding polypeptides having pamO activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 52 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 52, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 52. The invention does also include sequence variants of polypeptides of SEQ ID NO: 53 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 53. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 53 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 54, 55, 56, 57 and 58. Further variants can be deduced by the information provided by Figure 10 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 1 1 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 53, 54, 55, 56, 57 and 58. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 1 1 :

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 53 NO: 54 NO: 55 NO: 56 NO: 57 NO: 58

SEQ ID NO: 53 100% 95% 95% 93% 90% 90%

SEQ ID NO: 54 - 100% 91 % 88% 89% 88%

SEQ ID NO: 55 - - 100% 89% 88% 86%

SEQ ID NO: 56 - - - 100% 85% 84%

SEQ ID NO: 57 - - - - 100% 84%

SEQ ID NO: 58 - - - - - 100%

Further examples for polypeptides having pamO activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamO activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 258, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 257, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 257.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 257 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 257, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 257.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 258 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 258. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 258 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 259, 260, 261 , 262 and 263. Further variants can be deduced aligning the sequences of SEQ ID NO: 258, 259, 260, 261 , 262 and 263 to the information provided by Figure 10 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 1 1a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs 53, 258, 259, 260, 261 , 262 and 263. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 1 1a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 53 NO: 258 NO: 259 NO: 260 NO: 261 NO: 262 NO: 263

SEQ ID NO: 53 100% 93% 88% 88% 86% 84% 84%

SEQ ID NO: 258 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 259 - - 100% 89% 88% 85% 86%

SEQ ID NO: 260 - - - 100% 89% 87% 87%

SEQ ID NO: 261 - - - - 100% 86% 86%

SEQ ID NO: 262 - - - - - 100% 85%

SEQ ID NO: 263 - - - - - - 100%

Polypeptides having pamO activity which are derived from other metabolic pathways

Further examples for polypeptides having pamO activity are polypeptides being obtainable from Streptomyces fulvissimus and their variants.

Thus, the invention provides for polypeptides having pamOB activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 384, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 383, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 383.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 383 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 383, using genomic DNA or cDNA of Streptomyces fulvissimus as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 383. The invention does also include sequence variants of polypeptides of SEQ ID NO: 384 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 384. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 384 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 385, 386, 387, 388 and 389. Further variants can be deduced aligning the sequences of SEQ ID NO: 384, 385, 386, 387, 388 and 389 to the information provided by Figure 24 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 1 1 b shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 384, 385, 386, 387, 388 and 389. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 1 1b:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 384 NO: 385 NO: 386 NO: 387 NO: 388 NO: 389

SEQ ID NO: 384 100% 95% 95% 93% 90% 90%

SEQ ID NO: 385 - 100% 91 % 89% 86% 86%

SEQ ID NO: 386 - - 100% 88% 86% 86%

SEQ ID NO: 387 - - - 100% 86% 85%

SEQ ID NO: 388 - - - - 100% 85%

SEQ ID NO: 389 - - - - - 100%

Further examples for polypeptides having pamO activity are polypeptides being obtainable from Streptomyces griseus and their variants.

Thus, the invention provides for polypeptides having pamO activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 391 , and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 390, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 390.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 390 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 390, using genomic DNA or cDNA of Streptomyces griseus as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 390. The invention does also include sequence variants of polypeptides of SEQ ID NO: 391 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 391. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 391 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 392, 393, 394, 395 and 396. Further variants can be deduced aligning the sequences of SEQ ID NO: 391 , 392, 393, 394, 395 and 396 to the information provided by Figure 24 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 1 1c shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 391 , 392, 393, 394, 395 and 396. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 1 1c:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 391 NO: 392 NO: 393 NO: 394 NO: 395 NO: 396

SEQ ID NO: 391 100% 95% 95% 93% 90% 90%

SEQ ID NO: 392 - 100% 91 % 88% 87% 85%

SEQ ID NO: 393 - - 100% 88% 87% 87%

SEQ ID NO: 394 - - - 100% 85% 88%

SEQ ID NO: 395 - - - - 100% 85%

SEQ ID NO: 396 - - - - - 100%

Further examples for polypeptides having pamO activity are polypeptides being obtainable from Streptomyces sp. SolWspMP and their variants.

Thus, the invention provides for polypeptides having pamO activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 398, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 397, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 397.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 397 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 397, using genomic DNA or cDNA of Streptomyces sp. SolWspMP as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 397. The invention does also include sequence variants of polypeptides of SEQ ID NO: 398 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 398. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 398 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 399, 400, 401 , 402 and 403. Further variants can be deduced aligning the sequences of SEQ ID NO: 398, 399, 400, 401 , 402 and 403 to the information provided by Figure 24 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 1 1 d shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 398, 399, 400, 401 , 402 and 403. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 1 1d:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 398 NO: 399 NO: 400 NO: 401 NO: 402 NO: 403

SEQ ID NO: 398 100% 95% 95% 93% 90% 90%

SEQ ID NO: 399 - 100% 92% 88% 88% 86%

SEQ ID NO: 400 - - 100% 88% 87% 86%

SEQ ID NO: 401 - - - 100% 85% 86%

SEQ ID NO: 402 - - - - 100% 84%

SEQ ID NO: 403 - - - - - 100%

Further examples for polypeptides having pamO activity are polypeptides being obtainable from Streptomyces sp. W007 and their variants.

Thus, the invention provides for polypeptides having pamO activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 405, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 404, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 404.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 404 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 404, using genomic DNA or cDNA of Streptomyces sp. W007 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 404. The invention does also include sequence variants of polypeptides of SEQ ID NO: 405 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 405. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 405 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 406, 407, 408, 409 and 410. Further variants can be deduced aligning the sequences of SEQ ID NO: 405, 406, 407, 408, 409 and 410 to the information provided by Figure 24 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 1 1e shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 405, 406, 407, 408, 409 and 410. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 1 1 e:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 405 NO: 406 NO: 407 NO: 408 NO: 409 NO: 410

SEQ ID NO: 405 100% 95% 95% 93% 90% 90%

SEQ ID NO: 406 - 100% 90% 88% 86% 87%

SEQ ID NO: 407 - - 100% 89% 88% 86%

SEQ ID NO: 408 - - - 100% 86% 85%

SEQ ID NO: 409 - - - - 100% 85%

SEQ ID NO: 410 - - - - - 100%

Further examples for polypeptides having pamO activity are chimeric polypeptides and polynucleotides comprising polypeptide and polynucleotide sequences obtainable from Streptomyces californicus. Also comprised are polypeptides having pamO activity and the respective encoding polynucleotides being obtainable from Streptomyces californicus.

Thus, the invention provides for polypeptides having pamO activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 412, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 41 1 , or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 41 1.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 41 1 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 41 1 , using genomic DNA or cDNA of Streptomyces californicus as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 41 1.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 412 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 412. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 412 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 413, 414, 415, 416 and 417. Further variants can be deduced aligning the sequences of SEQ ID NO: 412, 413, 414, 415, 416 and 417 to the information provided by Figure 24 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 1 1f shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 412, 413, 414, 415, 416 and 417. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 1 1 f:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 412 NO: 413 NO: 414 NO: 415 NO: 416 NO: 417

SEQ ID NO: 412 100% 95% 95% 93% 90% 90%

SEQ ID NO: 413 - 100% 91 % 90% 87% 86%

SEQ ID NO: 414 - - 100% 89% 87% 86%

SEQ ID NO: 415 - - - 100% 85% 86%

SEQ ID NO: 416 - - - - 100% 84%

SEQ ID NO: 417 - - - - - 100%

Table 1 1g shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs 53, 258, 384, 391 , 398, 405 and 412. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 1 1g:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 53 NO: 258 NO: 384 NO: 391 NO: 398 NO: 405 NO: 412

SEQ ID NO: 53 100% 93% 57% 59% 57% 57% 58%

SEQ ID NO: 258 - 100% 59% 59% 59% 59% 59%

SEQ ID NO: 384 - - 100% 91 % 92% 92% 88%

SEQ ID NO: 391 - - - 100% 90% 98% 88%

SEQ ID NO: 398 - - - - 100% 90% 86%

SEQ ID NO: 405 - - - - - 100% 89%

SEQ ID NO: 412 - - - - - - 100%

Polypeptides having pamK activity:

A polypeptide having pamK activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 60, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 59, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 60. Preferably it is the same promoter.

The invention provides for polypeptides having pamK activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 59, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 59.

Preferably the polynucleotides encoding polypeptides having pamK activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 59 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 59, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 59.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 60 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 60. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 60 with other amino acids, preferably using conservative amino acid substitutions. Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 61 , 62, 63, 64 and 65. Further variants can be deduced by the information provided by Figure 1 1 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 12 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 60, 61 , 62, 63, 64 and 65. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 12:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 60 NO: 61 NO: 62 NO: 63 NO: 64 NO: 65

SEQ ID NO: 60 100% 95% 95% 93% 90% 90%

SEQ ID NO: 61 - 100% 91 % 90% 86% 88%

SEQ ID NO: 62 - - 100% 90% 89% 87%

SEQ ID NO: 63 - - - 100% 85% 84%

SEQ ID NO: 64 - - - - 100% 83%

SEQ ID NO: 65 - - - - - 100%

Further examples for polypeptides having pamK activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamK activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 265, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 264, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 264.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 264 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 264, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 264.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 265 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 265. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 265 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 266, 267, 268, 269 and 270. Further variants can be deduced aligning the sequences of SEQ ID NO: 265, 266, 267, 268, 269 and 270 to the information provided by Figure 1 1 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 12a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 60, 265, 266, 267, 268, 269 and 270. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 12a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 60 NO: 265 NO: 266 NO: 267 NO: 268 NO: 269 NO: 270

SEQ ID NO: 60 100% 95% 90% 90% 88% 85% 85% SEQ ID NO: 265 100% 95% 95% 93% 90% 90% SEQ ID NO: 266 100% 90% 89% 88% 86% SEQ ID NO: 267 100% 91 % 88% 89% SEQ ID NO: 268 100% 87% 85% SEQ ID NO: 269 100% 84% SEQ ID NO: 270 100%

Polypeptides having pamJ activity:

A polypeptide having pamJ activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 67, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 66, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 67. Preferably it is the same promoter.

The invention provides for polypeptides having pamJ activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 66. Preferably the polynucleotides encoding polypeptides having pamJ activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 66 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 66, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 66.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 67 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 67. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 67 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 68, 69, 70, 71 and 72. Further variants can be deduced by the information provided by Figure 12 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 13 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 67, 68, 69, 70, 71 and 72. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 13:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 67 NO: 68 NO: 69 NO: 70 NO: 71 NO: 72

SEQ ID NO: 67 100% 95% 95% 93% 90% 90%

SEQ ID NO: 68 - 100% 91 % 89% 91 % 87%

SEQ ID NO: 69 - - 100% 90% 86% 89%

SEQ ID NO: 70 - - - 100% 86% 87%

SEQ ID NO: 71 - - - - 100% 86%

SEQ ID NO: 72 - - - - - 100%

Further examples for polypeptides having pamJ activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants. Thus, the invention provides for polypeptides having pamJ activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 272, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 271 , or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 271.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 271 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 271 , using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 271.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 272 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 272. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 272 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 273, 274, 275, 276 and 277. Further variants can be deduced aligning the sequences of SEQ ID NO: 272, 273, 274, 275, 276 and 277 to the information provided by Figure 12 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 13a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 67, 272, 273, 274, 275, 276 and 277. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 13a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 67 NO: 272 NO: 273 NO: 274 NO: 275 NO: 276 NO: 277

SEQ ID NO: 67 100% 92% 87% 88% 85% 84% 84%

SEQ ID NO: 272 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 273 - - 100% 91 % 89% 86% 87%

SEQ ID NO: 274 - - - 100% 90% 87% 86%

SEQ ID NO: 275 - - - - 100% 87% 84%

SEQ ID NO: 276 - - - - - 100% 84%

SEQ ID NO: 277 - - - - - - 100%

Polypeptides having pamM activity:

A polypeptide having pamM activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 74, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 73, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 74. Preferably it is the same promoter.

The invention provides for polypeptides having pamM activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 73, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 73. Preferably the polynucleotides encoding polypeptides having pamM activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 73 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 73, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ I D NO: 73.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 74 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 74. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 74 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 75, 76, 77, 78 and 79. Further variants can be deduced by the information provided by Figure 13 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 14 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 74, 75, 76, 77, 78 and 79. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62 Table 14:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 74 NO: 75 NO: 76 NO: 77 NO: 78 NO: 79

SEQ ID NO: 74 100% 95% 95% 93% 90% 90%

SEQ ID NO: 75 - 100% 90% 89% 86% 87%

SEQ ID NO: 76 - - 100% 89% 86% 86%

SEQ ID NO: 77 - - - 100% 87% 86%

SEQ ID NO: 78 - - - - 100% 85%

SEQ ID NO: 79 - - - - - 100%

Further examples for polypeptides having pamM activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants. Thus, the invention provides for polypeptides having pamM activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 279, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 278, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 278.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 278 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 278, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 278. The invention does also include sequence variants of polypeptides of SEQ ID NO: 279 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ I D NO: 279. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 279 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 280, 281 , 282, 283 and 284. Further variants can be deduced aligning the sequences of SEQ ID NO: 279, 280, 281 , 282, 283 and 284 to the information provided by Figure 13 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 14a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 74, 279, 280, 281 , 282, 283 and 284. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62. Table 14a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 74 NO: 279 NO: 280 NO: 281 NO: 282 NO: 283 NO: 284

SEQ ID NO: 74 100% 85% 81 % 81 % 80% 76% 77%

SEQ ID NO: 279 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 280 - - 100% 93% 89% 87% 88%

SEQ ID NO: 281 - - - 100% 89% 87% 87%

SEQ ID NO: 282 - - - - 100% 86% 87%

SEQ ID NO: 283 - - - - - 100% 84%

SEQ ID NO: 284 - - - - - - 100%

Polypeptides having pamN activity:

A polypeptide having pamN activity is a polypeptide being able to complement the activity of a polypeptide of SEQ I D NO: 81 , or a polypeptide being expressed from a polynucleotide sequence of SEQ I D NO: 80, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 81 . Preferably it is the same promoter. The invention provides for polypeptides having pamN activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ I D NO: 81 , and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 80, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a complement of SEQ ID NO: 80.

Preferably the polynucleotides encoding polypeptides having pamN activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 80 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 80, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 80. The invention does also include sequence variants of polypeptides of SEQ ID NO: 81 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 81. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 81 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 82, 83, 84, 85 and 86. Further variants can be deduced by the information provided by Figure 14 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 15 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 81 , 82, 83, 84, 85 and 86. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 15:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 81 NO: 82 NO: 83 NO: 84 NO: 85 NO: 86

SEQ ID NO: 81 100% 95% 95% 93% 90% 90%

SEQ ID NO: 82 - 100% 90% 89% 85% 88%

SEQ ID NO: 83 - - 100% 90% 87% 89%

SEQ ID NO: 84 - - - 100% 87% 86%

SEQ ID NO: 85 - - - - 100% 84%

SEQ ID NO: 86 - - - - - 100%

Further examples for polypeptides having pamN activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants. Thus, the invention provides for polypeptides having pamN activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 286, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 285, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 285.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 285 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 285, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 285.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 286 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 286. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 286 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 287, 288, 289, 290 and 291 . Further variants can be deduced aligning the sequences of SEQ ID NO: 286, 287, 288, 289, 290 and 291 to the information provided by Figure 14 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 15a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 81 , 286, 287, 288, 289, 290 and 291 . The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62. Table 15a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 81 NO: 286 NO: 287 NO: 288 NO: 289 NO: 290 NO: 291

SEQ ID NO: 81 100% 92% 88% 87% 85% 83% 84% SEQ ID NO: 286 100% 95% 95% 93% 90% 90% SEQ ID NO: 287 100% 90% 88% 87% 87% SEQ ID NO: 288 100% 89% 88% 89% SEQ ID NO: 289 100% 85% 87% SEQ ID NO: 290 100% 85% SEQ ID NO: 291 100%

Polypeptides having pamL activity:

A polypeptide having pamL activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 88, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 87, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 88. Preferably it is the same promoter.

The invention provides for polypeptides having pamL activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 87.

Preferably the polynucleotides encoding polypeptides having pamL activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 87 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 87, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 87. The invention does also include sequence variants of polypeptides of SEQ ID NO: 88 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 88. These sequence variants can be obtained by substitution of one or more amino acids of SEQ I D NO: 88 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 89, 90, 91 , 92 and 93. Further variants can be deduced by the information provided by Figure 15 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 16 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 88, 89, 90, 91 , 92 and 93. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 16:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 88 NO: 89 NO: 90 NO: 91 NO: 92 NO: 93

SEQ ID NO: 88 100% 95% 95% 93% 90% 90%

SEQ ID NO: 89 - 100% 90% 89% 86% 88%

SEQ ID NO: 90 - - 100% 89% 87% 87%

SEQ ID NO: 91 - - - 100% 87% 86%

SEQ ID NO: 92 - - - - 100% 83%

SEQ ID NO: 93 - - - - - 100% Further examples for polypeptides having pamL activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamL activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 293, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 292, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 292.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 292 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 292, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 292.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 293 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 293. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 293 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 294, 295, 296, 297 and 298. Further variants can be deduced aligning the sequences of SEQ ID NO: 293, 294, 295, 296, 297 and 298 to the information provided by Figure 15 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 15a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 88, 293, 294, 295, 296, 297 and 298. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 16a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 88 NO: 293 NO: 294 NO: 295 NO: 296 NO: 297 NO: 298

SEQ ID NO: 88 100% 91 % 91 % 86% 85% 82% 82%

SEQ ID NO: 293 - 100% 100% 95% 93% 90% 90%

SEQ ID NO: 294 - - 100% 95% 93% 90% 90%

SEQ ID NO: 295 - - - 100% 89% 87% 87%

SEQ ID NO: 296 - - - - 100% 87% 85%

SEQ ID NO: 297 - - - - - 100% 84%

SEQ ID NO: 298 - - - - - - 100% Polypeptides having pamX activity:

A polypeptide having pamX activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 95, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 94, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ I D NO: 95. Preferably it is the same promoter.

The invention provides for polypeptides having pamX activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 95, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 94, or are expressed from a polynucleotide which hybridises under medium stringency

hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 94. Preferably the polynucleotides encoding polypeptides having pamX activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 94 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 94, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 94.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 95 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 95. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 95 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 96, 97, 98, 99 and 100. Further variants can be deduced by the information provided by Figure 16 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 17 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 95, 96, 97, 98, 99 and 100. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite, #GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 17:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 95 NO: 96 NO: 97 NO: 98 NO: 99 NO: 100

SEQ ID NO: 95 100% 95% 95% 93% 90% 90%

SEQ ID NO: 96 - 100% 91 % 90% 86% 88%

SEQ ID NO: 97 - - 100% 90% 87% 87%

SEQ ID NO: 98 - - - 100% 86% 85%

SEQ ID NO: 99 - - - - 100% 85%

SEQ ID NO: 100 - - - - - 100% Further examples for polypeptides having pamX activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamX activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 300, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 299, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 299.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 299 per primer, wherein the primers are capable to produce a PCR fragment of SEQ I D NO: 299, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 299.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 300 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 300. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 300 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 301 , 302, 303, 304 and 305. Further variants can be deduced aligning the sequences of SEQ ID NO: 300, 301 , 302, 303, 304 and 305 to the information provided by Figure 16 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 17a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 95, 300, 301 , 302, 303, 304 and 305. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 17a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 95 NO: 300 NO: 301 NO: 302 NO: 303 NO: 304 NO: 305

SEQ ID NO: 95 100% 95% 90% 90% 88% 85% 85%

SEQ ID NO: 300 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 301 - - 100% 92% 89% 87% 88%

SEQ ID NO: 302 - - - 100% 90% 87% 87%

SEQ ID NO: 303 - - - - 100% 87% 86%

SEQ ID NO: 304 - - - - - 100% 85%

SEQ ID NO: 305 - - - - - - 100% Polypeptides having pamY activity:

A polypeptide having pamY activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 102, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 101 , in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 102. Preferably it is the same promoter.

The invention provides for polypeptides having pamY activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 101 , or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 101.

Preferably the polynucleotides encoding polypeptides having pamY activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 101 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 101 , using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 101.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 102 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 102. These sequence variants can be obtained by substitution of one or more amino acids of SEQ I D NO: 102 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 103, 104, 105, 106 and 107. Further variants can be deduced by the information provided by Figure 17 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 18 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 102, 103, 104, 105, 106 and 107. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 18:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 102 NO: 103 NO: 104 NO: 105 NO: 106 NO: 107

SEQ ID NO 102 100% 95% 95% 93% 90% 90%

SEQ ID NO 103 - 100% 92% 88% 87% 85%

SEQ ID NO 104 - - 100% 89% 86% 86%

SEQ ID NO 105 - - - 100% 85% 84%

SEQ ID NO 106 - - - - 100% 85%

SEQ ID NO 107 - - - - - 100%

Further examples for polypeptides having pamY activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamY activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 307, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 306, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 306.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 306 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 306, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 306.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 307 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 307. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 307 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 308, 309, 310, 31 1 and 312. Further variants can be deduced aligning the sequences of SEQ ID NO: 307, 308, 309, 310, 31 1 and 312 to the information provided by Figure 17 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 18a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 102, 307, 308, 309, 310, 31 1 and 312. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 18a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 102 NO: 307 NO: 308 NO: 309 NO: 310 NO: 31 1 NO: 312

SEQ ID NO: 102 100% 94% 89% 89% 86% 84% 84%

SEQ ID NO: 307 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 308 - - 100% 90% 89% 86% 88%

SEQ ID NO: 309 - - - 100% 89% 87% 87%

SEQ ID NO: 310 - - - - 100% 86% 85%

SEQ ID NO: 31 1 - - - - - 100% 84%

SEQ ID NO: 312 - - - - - - 100%

Polypeptides having pamS activity:

A polypeptide having pamS activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 109, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 108, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 109. Preferably it is the same promoter.

The invention provides for polypeptides having pamS activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 108, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 108. Preferably the polynucleotides encoding polypeptides having pamS activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 108 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 108, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 108.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 109 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 109. These sequence variants can be obtained by substitution of one or more amino acids of SEQ I D NO: 109 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 1 10, 1 1 1 , 1 12, 1 13 and 1 14. Further variants can be deduced by the information provided by Figure 18 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 19 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 109, 1 10, 1 1 1 , 1 12, 1 13 and 1 14. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 19:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 109 NO: 1 10 NO: 1 1 1 NO: 1 12 NO: 1 13 NO: 1 14

SEQ ID NO 109 100% 95% 95% 93% 90% 90%

SEQ ID NO 1 10 - 100% 91 % 92% 85% 87%

SEQ ID NO 1 1 1 - - 100% 89% 86% 87%

SEQ ID NO 1 12 - - - 100% 84% 85%

SEQ ID NO 1 13 - - - - 100% 84%

SEQ ID NO 1 14 - - - - - 100%

Further examples for polypeptides having pamS activity are chimeric polypeptides, comprising sequence elements obtainable from polypeptides having pamS activity of Streptomyces alboniger and sequence lements of polypeptides having pamS activity being obtainable from Streptomyces aurantiacus JA 4570. Examples, but not excluding others, of these chimeric polypeptides are provided below. Also comprised are polypeptides having pamS activity and polynucleotides encoding these being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Accordingly, the invention provides for polypeptides having pamS activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 314, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 313, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 108, preferably to a complement of SEQ ID NO: 313.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 108 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 108, using genomic DNA or cDNA of Streptomyces alboniger as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 108.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 314 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 314. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 314 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 315, 316, 317, 318 and 319. Further variants can be deduced aligning the sequences of SEQ ID NO: 306, 315, 316, 317, 318 and 319 to the information provided by Figure 18 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 19a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 109, 306, 315, 316, 317, 318 and 319. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 19a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 109 NO: 314 NO: 315 NO: 316 NO: 317 NO: 318 NO: 319

SEQ ID NO: 109 100% 94% 89% 89% 88% 85% 85% SEQ ID NO: 314 100% 95% 95% 93% 90% 90% SEQ ID NO: 315 100% 90% 91 % 87% 88% SEQ ID NO: 316 100% 89% 88% 87% SEQ ID NO: 317 100% 87% 86% SEQ ID NO: 318 100% 85% SEQ ID NO: 319 100%

Polypeptides having pamR1 activity: A polypeptide having pamR1 activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 1 16, or a polypeptide being expressed from a

polynucleotide sequence of SEQ ID NO: 1 15, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the complemented strain for the capability to produce pamamycin 607. A successful complementation is achieved, if the complemented strain produces pamamycin 607 on a higher level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 1 16. Preferably it is the same promoter.

The invention provides for polypeptides having pamR1 activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 1 15.

Preferably the polynucleotides encoding polypeptides having pamR1 activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 1 15, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 1 15.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 1 16 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 1 16. These sequence variants can be obtained by substitution of one or more amino acids of SEQ I D NO: 1 16 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 1 17, 1 18, 1 19, 120 and 121. Further variants can be deduced by the information provided by Figure 19 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 20 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 1 16, 1 17, 1 18, 1 19, 120 and 121 . The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 20:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 1 16 NO: 1 17 NO: 1 18 NO: 1 19 NO: 120 NO: 121

SEQ ID NO 1 16 100% 95% 95% 93% 90% 90%

SEQ ID NO 1 17 - 100% 90% 90% 89% 87%

SEQ ID NO 1 18 - - 100% 89% 87% 86%

SEQ ID NO 1 19 - - - 100% 87% 85%

SEQ ID NO 120 - - - - 100% 84%

SEQ ID NO 121 - - - - - 100% Further examples for polypeptides having pamR1 activity are polypeptides being

obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamR1 activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 321 , and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 320, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 320.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 320 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 320, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 320.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 321 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 321. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 321 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ I D NO: 322, 323, 324, 325 and 326. Further variants can be deduced aligning the sequences of SEQ ID NO: 321 , 322, 323, 324, 325 and 326 to the information provided by Figure 19 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 20a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 1 16, 321 , 322, 323, 324, 325 and 326. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 20a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 1 16 NO: 321 NO: 322 NO: 323 NO: 324 NO: 325 NO: 326

SEQ ID NO: 1 16 100% 90% 85% 85% 83% 82% 80%

SEQ ID NO: 321 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 322 - - 100% 90% 89% 87% 86%

SEQ ID NO: 323 - - - 100% 87% 89% 88%

SEQ ID NO: 324 - - - - 100% 86% 86%

SEQ ID NO: 325 - - - - - 100% 84%

SEQ ID NO: 326 - - - - - - 100% Polypeptides having pamH activity:

A polypeptide having pamH activity is a polypeptide being able to complement the activity of a polypeptide of SEQ I D NO: 123, or a polypeptide being expressed from a polynucleotide sequence of SEQ ID NO: 122, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the

complemented strain for the capability to produce pamamycin 607. A successful

complementation is achieved, if the complemented strain produces pamamycin 607 on a lower level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 123. Preferably it is the same promoter.

The invention provides for polypeptides having pamH activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123, and/or are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 122, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 122.

Preferably the polynucleotides encoding polypeptides having pamH activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 122 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 122, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ I D NO: 122.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 123 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ I D NO: 123. These sequence variants can be obtained by substitution of one or more amino acids of SEQ I D NO: 123 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 124, 125, 126, 127 and 128. Further variants can be deduced by the information provided by Figure 20 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 21 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 123, 124, 125, 126, 127 and 128. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62 Table 21 :

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 123 NO: 124 NO: 125 NO: 126 NO: 127 NO: 128

SEQ ID NO 123 100% 95% 95% 93% 90% 90%

SEQ ID NO 124 - 100% 91 % 90% 87% 86%

SEQ ID NO 125 - - 100% 90% 88% 86%

SEQ ID NO 126 - - - 100% 85% 88%

SEQ ID NO 127 - - - - 100% 85%

SEQ ID NO 128 - - - - - 100% Further examples for polypeptides having pamH activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamH activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 328, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 327, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 327.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 327 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 327, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 327.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 328 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ I D NO: 328. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 328 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 329, 330, 331 , 332 and 333. Further variants can be deduced aligning the sequences of SEQ ID NO: 328, 329, 330, 331 , 332 and 333 to the information provided by Figure 20 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 21a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 123, 328, 329, 330, 331 , 332 and 333. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN: 10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62. Table 21a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 123 NO: 328 NO: 329 NO: 330 NO: 331 NO: 332 NO: 333

SEQ ID NO: 123 100% 90% 85% 86% 83% 81 % 82% SEQ ID NO: 328 100% 95% 95% 93% 90% 90% SEQ ID NO: 329 100% 91 % 89% 88% 87% SEQ ID NO: 330 100% 89% 87% 87% SEQ ID NO: 331 100% 87% 87% SEQ ID NO: 332 100% 83% SEQ ID NO: 333 100% Polypeptides having pamR2 activity:

A polypeptide having pamR2 activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 130, or a polypeptide being expressed from a

polynucleotide sequence of SEQ ID NO: 129, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the complemented strain for the capability to produce pamamycin 607. A successful complementation is achieved, if the complemented strain produces pamamycin 607 on a lower level than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 130. Preferably it is the same promoter.

The invention provides for polypeptides having pamR2 activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 129, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 129.

Preferably the polynucleotides encoding polypeptides having pamR2 activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 129 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 129, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 129. The invention does also include sequence variants of polypeptides of SEQ ID NO: 130 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 130. These sequence variants can be obtained by substitution of one or more amino acids of SEQ I D NO: 130 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 131 , 132, 133, 134 and 135. Further variants can be deduced by the information provided by Figure 21 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 22 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 130, 131 , 132, 133, 134 and 135. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62

Table 22:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 130 NO: 131 NO: 132 NO: 133 NO: 134 NO: 135

SEQ ID NO 130 100% 95% 95% 93% 90% 90%

SEQ ID NO 131 - 100% 90% 90% 89% 88%

SEQ ID NO 132 - - 100% 90% 88% 87%

SEQ ID NO 133 - - - 100% 84% 84%

SEQ ID NO 134 - - - - 100% 83%

SEQ ID NO 135 - - - - - 100%

Further examples for polypeptides having pamR2 activity are polypeptides being

obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamR2 activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 335, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 334, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 334.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 334 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 334, using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 334.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 335 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 335. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 335 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 336, 337, 338, 339 and 340. Further variants can be deduced aligning the Sequences of SEQ ID NO: 335, 336, 337, 338, 339 and 340 to the information provided by Figure 21 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions.

Table 22a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 130, 335, 336, 337, 338, 339 and 340. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62.

Table 22a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID

NO: 130 NO: 335 NO: 336 NO: 337 NO: 338 NO: 339 NO: 340

SEQ ID NO: 130 100% 91 % 86% 86% 84% 81 % 83%

SEQ ID NO: 335 - 100% 95% 95% 93% 90% 90%

SEQ ID NO: 336 - - 100% 91 % 90% 87% 87%

SEQ ID NO: 337 - - - 100% 88% 88% 87%

SEQ ID NO: 338 - - - - 100% 85% 87%

SEQ ID NO: 339 - - - - - 100% 86%

SEQ ID NO: 340 - - - - - - 100%

Polypeptides having pamW activity:

A polypeptide having pamW activity is a polypeptide being able to complement the activity of a polypeptide of SEQ ID NO: 137, or a polypeptide being expressed from a

polynucleotide sequence of SEQ ID NO: 136, in a Streptomyces alboniger strain available from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures under the DMS Nr. 40043. Complementation of the activity of such polypeptide can be tested by deleting the polynucleotide sequence encoding such polypeptide from the genome of Streptomyces alboniger DMS Nr. 40043 and transforming the resulting strain with an expression cassette capapble to express the polypeptide to be tested in such strain and testing the complemented strain for the capability to export pamamycin 607. A successful complementation is achieved, if the complemented strain exportes pamamycin 607 on a higher level from its cells to the growth medium than the strain comprising the deletion under growth conditions under which the unmodified Streptomyces alboniger DMS Nr. 40043 produces pamamycin 607, wherein the complemented strain uses a promoter to express the polypeptide to be tested, which has a similar or higher expression level under such growth conditions than the promoter of Streptomyces alboniger DMS Nr. 40043 driving expression of the polypeptide of SEQ ID NO: 137.

The invention provides for polypeptides having pamW activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, and/or are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 136, or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 136.

Preferably the polynucleotides encoding polypeptides having pamW activity can be obtained from an organism producing at least one of the pamamycins of Formula (I), preferably producing pamamycin 607. Preferreably such polynucleotides are obtained from organisms of the genus Streptomyces or obtained via sequence database searches comprising sequence information of organisms of the genus Streptomyces.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 136 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 136, using genomic DNA or cDNA of Streptomyces alboniger DMS Nr. 40043 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 136.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 137 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 137. These sequence variants can be obtained by substitution of one or more amino acids of SEQ I D NO: 137 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 138, 139, 140, 141 and 142. Further variants can be deduced by the information provided by Figure 22 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 23 shows the % sequence identity of pairwise sequence alignments of SEQ ID NO: 137, 138, 139, 140, 141 and 142. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62 Table 23:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 137 NO: 138 NO: 139 NO: 140 NO: 141 NO: 142

SEQ ID NO 137 100% 95% 95% 93% 90% 90%

SEQ ID NO 138 - 100% 90% 89% 86% 87%

SEQ ID NO 139 - - 100% 88% 86% 86%

SEQ ID NO 140 - - - 100% 85% 86%

SEQ ID NO 141 - - - - 100% 82%

SEQ ID NO 142 - - - - - 100% Further examples for polypeptides having pamW activity are polypeptides being obtainable from Streptomyces aurantiacus JA 4570 and their variants.

Thus, the invention provides for polypeptides having pamW activity, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 342, and/or

are expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 341 , or are expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, preferably high stringency hybridisation conditions, to a

complement of SEQ ID NO: 341.

A preferred method to obtain such polynucleotides is to perform PCR on genomic DNA or cDNA of pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 341 per primer, wherein the primers are capable to produce a PCR fragment of SEQ ID NO: 341 , using genomic DNA or cDNA of Streptomyces aurantiacus JA 4570 as PCR template. Preferred primers are capable to produce a full length fragment of SEQ ID NO: 341.

The invention does also include sequence variants of polypeptides of SEQ ID NO: 342 which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical SEQ ID NO: 342. These sequence variants can be obtained by substitution of one or more amino acids of SEQ ID NO: 342 with other amino acids, preferably using conservative amino acid substitutions.

Examples of such sequence variants, but not excluding others, are polypeptides of SEQ ID NO: 343, 344, 345, 346 and 347. Further variants can be deduced aligning the sequences of SEQ ID NO: 342, 343, 344, 345, 346 and 347 to the information provided by Figure 22 and its description provided above. Preferred variants comprise not more than 15, 14, 13, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, additions or deletions. Table 23a shows the % sequence identity of pairwise sequence alignments of SEQ ID NOs: 137, 342, 343, 344, 345, 346 and 347. The parameters used for the pairwise sequence alignments are: Needleman-Wunsch algorithm of the EMBOSS Software Suite,

#GAPMETHOD: NOGAPS, #GAPOPEN:10, GAPEXTEND: 0,5, MATRIX: EBLOSUM62. Table 23a:

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 137 NO: 342 NO: 343 NO: 344 NO: 345 NO: 346 NO: 347

SEQ ID NO: 137 100% 89% 84% 84% 82% 80% 80% SEQ ID NO: 342 100% 95% 95% 93% 90% 90% SEQ ID NO: 343 100% 91 % 89% 86% 86% SEQ ID NO: 344 100% 89% 87% 86% SEQ ID NO: 345 100% 86% 84% SEQ ID NO: 346 100% 83% SEQ ID NO: 347 100% Variants of the polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS, pamR1 , pamR2, pamW or pamH activity can be obtained by techniques known to a person skilled in the art from pamamycin producing organisms. Preferred pamamycin producing microorganism are: Streptomyces alboniger ATCC 12461 Streptomyces alboniger DSM 40043, Streptomyces alboniger IF012738, Streptomyces aurantiacus ATCC 19822 and Streptomyces

kitasatoensis J CM 5001.

Further, polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS, pamR1 , pamR2, pamW or pamH activity and the polynucleotides encoding these polypeptides can be used to identify pamamycin producing microorganisms, improve the production of pamamycin by such microorganisms and/or to construct new pamamycin producing organisms.

Accordingly, the invention comprises recombinant microorganism comprising at least one up-regulated activity selected from pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS. Such recombinant microorganism may be man-made mutants of pamamycin producing organism, which comprise mutations enhancing at least one of those activites in comparison to the un- mutated strain. Such mutations can comprise any mutation which enhances such activities, for example but not excluding others, duplications of the respective expression cassette for the polypeptides having such activities, promoter mutations enhancing production of such polypeptides, mutations enhancing the activity or stability of such polypeptides, mutations deleting or reducing feed-back inhibitons of such polypeptides, deletions or defect- mutations of repressors of such activities or of the expression of the respective polypeptides having such activites. Alternative mutants may have an up-regulated pamR1 and/or pamW activity, or a down-regulated pamR2 and/or pamH activity or a combination of those, or a combination of these with one or more mutations leading to at least one up-regulated pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, or pamS activity.

A further way to improve or modulate pamamycin production in microorganisms is to construct recombinant microorganisms by transformation of pamamycin producing organisms with recombinant expression cassettes encoding polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY or pamS activity, or by replacing promoter, terminator or polypeptiptide encoding regions of polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY or pamS activity with other versions, which result in an up-regulated pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY or pamS activity activity in such recombinant microorganisms.

Another way to construct recombinant microorganisms of the invention is to provide a microorganism which naturally does not produce pamamycin with one or more expression cassettes for polypeptides, so that the resulting microorganism has at least one expression cassette coding for a polypeptide for each one of pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity. An individual expression cassette may encode only one polypeptide having one of the above mentioned activities, but may also have an operon like structure, so that it encodes more than one polypeptide having the same or different activities selected from the ones described above.

Accordingly, the invention encompases recombinant microorganisms comprising:

a) at least one polypeptide having pamC activity, and

b) at least one polypeptide having pamG activity, and

c) at least one polypeptide having pamF activity, and

d) at least one polypeptide having pamA activity, and

e) at least one polypeptide having pamB activity, and

f) at least one polypeptide having pamD activity, and

g) at least one polypeptide having pamE activity, and

h) at least one polypeptide having pamO activity, and

i) at least one polypeptide having pamK activity, and

j) at least one polypeptide having pamJ activity, and

k) at least one polypeptide having pamM activity, and

I) at least one polypeptide having pamN activity, and

m) at least one polypeptide having pamL activity, and

n) at least one polypeptide having pamX activity, and

o) at least one polypeptide having pamY activity, and

P) at least one polypeptide having pamS activity,

wherein at least one of those activities a) to p) is up-regulated.

Miroorganisms which have at least one polypeptide encoding region or expression cassette for each one of the polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity, being either naturally pamamycin producing microorganisms, or microorganisms which are recombinant for at least one of those expression cassettes, can be provided with recombinant expression cassettes encoding polypeptides having pamR1 or pamW actitiy, or with at least one expression casette for a polypeptide having pamR1 activity and at least one expression cassette for a polypeptide having pamW actitity. Alternatively or in addition to providing a naturally pamamycin producing microorganism, or its man-made mutant, or microorganism being recombinant for an expression cassette for at least one polypeptide having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY or pamS activity with further expression cassettes for at least one of those expression cassette, such naturally pamamycin producing microorganisms may be provided with recombinant polynucleotides wich lead to a reduced expression of polypeptides having pamR2 or pamH actitity, or which lead to a reduced activity or stability of the polypeptides having pamR2 or pamH actitity. Accordingly, the invention encompases recombinant microorganisms comprising:

at least one polypept de having pamC activity, and

at least one polypept de having pamG activity, and

at least one polypept de having pamF activity, and

at least one polypept de having pamA activity, and

at least one polypept de having pamB activity, and

at least one polypept de having pamD activity, and

at least one polypept de having pamE activity, and

at least one polypept de having pamO activity, and

at least one polypept de having pamK activity, and

at least one polypept de having pamJ activity, and

at least one polypept de having pamM activity, and

at least one polypept de having pamN activity, and

at least one polypept de having pamL activity, and

at least one polypept de having pamX activity, and

at least one polypept de having pamY activity, and

at least one polypept de having pamS activity, and comprising at least one of q), r), s), t), u) or v), being

q) an up-regulated polypeptide having pamR1 activity,

r) a down-regulated polypeptide having pamR2 activity,

s) an up-regulated polypeptide having pamW activity,

t) a down-regulated polypeptide having pamH activity,

u) a combination of at least two of q), r) s) and t),

v) at least one up-regulated activities of a) to p) and at least one of q) to u).

The polypeptides of a) to p), or of a) to u) are preferably expressed from expression cassettes encoding at least one, or several of the polypeptides of a) to p) or of a) to u), expression cassettes producing mRNAs having an operon like structure.

The polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS, pamR1 , pamR2, pamW or pamH activity can be provided by the polypeptides and polynucleotides as described above, including their variants, in particular their variants listed in Tables 4, 4a, 5, 5a, 6, 6a, 7, 7a, 8, 8a to 8g, 9, 9a, 10, 10a, 1 1 , 1 1a to 1 1g, 12, 12a, 13, 13a, 14, 14a, 15, 15a, 16, 16a, 17, 17a, 18, 18a, 19, 19a, 20, 20a, 21 , 21a, 22, 22a, 23, 23a and polypeptides having at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

Accordingly, the invention provides for recombinant microorganisms, wherein at least one polypeptide having pamC activity, is provided by

a1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4 and/or 209, or a2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, 5, 6, 7, 8, 9, 209,

210, 21 1 , 212, 213 and/or 214, or

a3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3 and/or 208 or

a4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 3 and/or 208, or

a5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 3 and/or 208 per primer, or

a6) a polypeptide of a1) and/or a2) and comprising the amino acids L29, D32, S33, E44 and F47 using the alignment in Figure 3 as reference, or a7) a polypeptide of at least two of a1 ), a2), a3), a4), a5) and a6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamG activity, is provided by

b1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 and/or 216, or

b2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 , 12, 13, 14, 15, 16, 216, 217, 218, 219, 220 and/or 221 , or

b3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 10 and/or 215, or

b4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 10 and/or 215, or

b5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 10 and/or 215 per primer, or

b6) a polypeptide of b1) and/or b2) and comprising the amino acids P19, A64, C1 16, L139, D163, A167, H258, G287 and N288 using the alignment in Figure 4 as reference, or

b7) a polypeptide of at least two of b1 ), b2), b3), b4), b5) and b6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamF activity, is provided by c1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 18 and/or 223, or

c2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18, 19, 20, 21 , 22, 23,

223, 224, 225, 226, 227 and/or 228, or

c3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17 and/or 222, or

c4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 17 and/or 222, or

c5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 17 and/or 222 per primer, or

c6) a polypeptide of c1 ) and/or c2) and comprising the amino acids G16, A186,

G255, G261 , G366 and H267 using the alignment in Figure 5 as reference, or

c7) a polypeptide of at least two of c1 ), c2), c3), c4), c5) and c6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamA activity, is provided by

d) at least one polypeptide having pamA activity, is provided by

d1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25 and/or 230, or

d2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, 26, 27, 28, 29, 30, 230, 231 , 232, 233, 234 and/or 235, or

d3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 24 and/or 229, or

d4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 24 and/or 229, or

d5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 24 and/or 229 per primer, or

d6) a polypeptide of d1) and/or d2) and comprising the amino acids G21 ,

C191 , G258, G264, H333, G371 and H272 using the alignment in Figure 6 as reference, or

d7) a polypeptide of at least two of d 1 ), d2), d3), d4), d5) and d6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamB activity, is provided by

e1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 237, 349, 356, 363, 370 and/or 377, preferably to SEQ ID NO: 32, 237 and/or 349, or

e2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 33, 34, 35, 36, 37, 237, 238, 239, 240, 241 , 242, 249, 350, 351 , 352, 353, 354, 356, 357, 358, 359, 360, 361 , 363, 364, 365, 366, 367, 368, 369, 371 , 372, 373, 374, 375, 377, 378, 379, 380, 381 and/or 382, preferably to SEQ ID NO: 32, 33, 34, 35, 36, 37, 237, 238, 239, 240, 241 , 242, 249, 350, 351 , 352, 353 and/or

354, or

e3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, preferably to SEQ ID NO: 31 , 236 and/or 348, or

e4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, preferably of SEQ ID NO: 31 , 236 and/or 348, or

e5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, preferably of SEQ ID NO: 31 , 236 and/or 348 per primer, or

e6) a polypeptide of e1 ) and/or e2) and comprising the amino acids T2, G3,

P6, R22, A23, P59, L89, D218, G221 , N222, P260, P266, V270, G407, G409, E434, G436, G491 and at least two of G506, G508, G509 using the alignment in Figure 7 as reference, or

e7) a polypeptide of at least two of e1 ), e2), e3), e4), e5) and e6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamD activity, is provided by

f 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39 and/or 244, or

f2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, 40, 41 , 42, 43, 44, 244, 245, 246, 247, 248 and/or 249, or

f3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38 and/or 243, or

f4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 38 and/or 243, or

f5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ I D

NO: 38 and/or 243 per primer, or

f6) a polypeptide of f1) and/or f2) and comprising the amino acids P21 , P101 ,

C121 , H246, G275 and G308 using the alignment in Figure 8 as reference, or

f7) a polypeptide of at least two of f 1 ), f2), f3), f4), f5) and f6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamE activity, is provided by

g1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 46 and/or 251 , or

g2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46, 47, 48, 49, 50, 51 , 251 , 252, 253, 254, 255 and/or 256, or

g3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45 and/or 250, or

g4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 45 and/or 250, or

g5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 45 and/or 250 per primer, or

g6) a polypeptide of g1) and/or g2) and comprising the amino acids P25, P103,

C136, H270, G299 and G333 using the alignment in Figure 9 as reference, or

g7) a polypeptide of at least two of g 1 ), g2), g3), g4), g5) and g6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamO activity, is provided by

hi ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 53, 258, 384, 391 , 398, 405, preferably to SEQ I D NO: 53, 258, 384 and/or 412, more preferred to to SEQ I D NO: 53 or

h2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 53, 54, 55, 56, 57, 58,

384, 385, 386, 387, 388, 389, 391 , 392, 393, 394, 395, 396, 398, 399, 400, 401 , 402, 403, 405, 406, 407, 408, 409, 410, 412, 413, 414, 415, 416 and/or 417, preferably to SEQ I D NO: 53, 54, 55, 56, 57, 58, 384, 385, 386, 387, 388, 389, 391 , 392, 393, 394, 395 and or 396, , or h3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , preferably to SEQ I D NO: 52, 257 and/or 383, or

h4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ I D

NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , preferably to SEQ I D NO: 52, 257 and/or 383, or

h5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ I D

NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , preferably of SEQ I D NO: 52, 257 and/or 383, per primer, or

h6) a polypeptide of h i ) and/or h2) and comprising the amino acids G 1 1 , G 17, D61 , Y164, K168, P 194 and at least two of G256, G257 and G258 using the alignment in Figure 10 as reference, or

h7) a polypeptide of at least two of h 1 ), h2), h3), h4), h5) and h6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamK activity, is provided by

i 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 60 and/or 265, or

\2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 60, 61 , 62, 63, 64, 65, 265, 266, 267, 268, 269 and/or 270, or

13) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 59 and/or 264, or

14) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ I D

NO: 59 and/or 264, or

15) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 59 and/or 264 per primer, or

a polypeptide of i 1 ) and/or i2) and comprising the amino acids F33, C159, G314 and at least one of G346 and G348 using the alignment in Figure 1 1 as reference, or

a polypeptide of at least two of i 1 ) , i2), i3), i4), i5) and i6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamJ activity, is provided by

j1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67 and/or 272, or

j2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, 68, 69, 70, 71 , 72,

272, 273, 274, 275, 276 and/or 277, or

j3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66 and/or 271 , or

j4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 66 and/or 271 , or

j5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 66 and/or 271 per primer, or

j6) a polypeptide of j 1 ) and/or j2) and comprising the amino acids G14, G246,

G252, G256 and H257 using the alignment in Figure 12 as reference, or j7) a polypeptide of at least two of j 1 ) , j2), j3), j4), j5) and j6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamM activity, is provided by

k1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74 and/or 279, or

k2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, 75, 76, 77, 78, 79, 279, 280, 281 , 282, 283 and/or 284, or

k3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 73 and/or 278, or

k4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 73 and/or 278, or

k5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 73 and/or 278 per primer, or

k6) a polypeptide of k1 ) and/or k2) and comprising the amino acids G1 1 , V139,

G241 , G247, D291 , A319, Y384, K388, P414, G415, T419, G465, G473 and at least one of G472 and G473 using the alignment in Figure 13 as reference, or

k7) a polypeptide of at least two of k1 ), k2), k3), k4), k5) and k6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamN activity, is provided by

11 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 and/or 286, or

12) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , 82, 83, 84, 85, 86, 286, 287, 288, 289, 290 and/or 291 , or

13) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 80 and/or 285, or

14) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 80 and/or 285, or

15) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 80 and/or 285 per primer, or

16) a polypeptide of 11 ) and/or I2) and comprising the amino acids G16, G22, D66, Y158, K162, P188 and G189 using the alignment in Figure 143 as reference, or

17) a polypeptide of at least two of 11 ), I2), I3), I4), I5) and I6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamL activity, is provided by

ml ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88 and/or 293, or

m2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, 89, 90, 91 , 92, 93, 293, 294, 295, 296, 297 and/or 298, or

m3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87 and/or 292, or

m4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 87 and/or 292, or

m5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 87 and/or 292 per primer, or

m6) a polypeptide of ml ) and/or m2) and comprising the amino acids L82, V93,

T214, S215, G216, G219, K222, E352, G370, G394, Y410, T438, G439,

D430, R445, G454, E463, P520, P530, G535, K536 and at least one of

T217 and T218 using the alignment in Figure 15 as reference, or m7) a polypeptide of at least two of ml ), m2), m3), m4), m5) and m6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamX activity, is provided by

n1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 95 and/or 300, or

n2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 95, 96, 97, 98, 99, 100,

300, 301 , 302, 303, 304 and/or 305, or

n3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 94 and/or 299, or

n4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 94 and/or 299, or

n5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 94 and/or 299 per primer, or

n6) a polypeptide of n1) and/or n2) and comprising the amino acids D46, G57,

G1 13, H147, D241 , K271 , G276, T308 and at least one of G40 and G41 using the alignment in Figure 16 as reference, or

n7) a polypeptide of at least two of n 1 ), n2), n3), n4), n5) and n6).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamY activity, is provided by 01 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102 and/or 307, or

02) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102, 103, 104, 105,

106, 107, 307, 308, 309, 310, 31 1 and/or 312, or

03) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 101 and/or 306, or

o4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 101 and/or 306, or

05) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 101 and/or 306 per primer, or

06) a polypeptide of o1 ) and/or o2) and comprising the amino acids G67, G69, D86, and at least one of G 153 and G 154, and at least one of G243 and G244 using the alignment in Figure 17 as reference, or

o7) a polypeptide of at least two of o1 ), o2), o3), o4), o5) and 06).

Further provided are recombinant microorganisms, wherein at least one polypeptide having pamS activity, is provided by

p1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 109 and/or 314, or

p2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, 1 10, 1 1 1 , 1 12, 1 13, 1 14, 314, 315, 316, 317, 318 and/or 319, or

p3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 108 and/or 313, or

p4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 108 and/or 313, or

p5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 108 and/or 313 per primer, or

p6) a polypeptide of p1) and/or p2) and comprising the amino acids N41 , F76,

G79, D81 , G1 16, G122, D130, G148, G187, K221 and at least two of G 120 G121 and G122 using the alignment in Figure 18 as reference, or p7) a polypeptide of at least two of p1 ), p2), p3), p4), p5) and p6);

Further provided are recombinant microorganisms, wherein at least one polypeptide

having pamR1 activity, is provided by

q1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16 and/or 321 , or

q2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17, 1 18, 1 19, 120, 121 , 321 , 322, 323, 324, 325 and/or 326, or

q3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15 and/or 320, or

q4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ I D

NO: 1 15 and/or 320, or

q5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 and/or 320 per primer, or

q6) a polypeptide of q 1 ) and/or q2) and comprising the amino acids K105, K190 and at least one of E153 and E154 using the alignment in Figure 19 as reference, or

q7) a polypeptide of at least two of q1 ), q2), q3), q4), q5) and q6);

Further provided are recombinant microorganisms, wherein at least one polypeptide

having pamR2 activity, is provided by

r1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130 and/or 335, or

r2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, 131 , 132, 133, 134, 135, 335, 336, 337, 338, 339 and/or 340, or

r3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 129 and/or 334, or

r4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 129 and/or 334, or

r5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 129 and/or 334 per primer, or

a polypeptide of r1 ) and/or r2) and comprising the amino acids E3, W19, A59, Y70, G169, P207, F226, L230, G236 at least one of A40 and A41 and at least one of G48 and G49 and at least one of L79 and L80 using the alignment in Figure 21 as reference, or

a polypeptide of at least two of r1 ), r2), r3), r4), r5) and r6);

Further provided are recombinant microorganisms, wherein at least one polypeptide

having pamW activity, is provided by

s1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137 and/or 342, or

s2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, 138, 139, 140, 141 , 142, 342, 343, 344, 345, 346 and/or 347, or

s3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 136 and/or 341 , or

s4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 136 and/or 341 , or

s5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 136 and/or 341 per primer, or

s6) a polypeptide of s1 ) and/or s2) and comprising the amino acids D92, G95 and G131 using the alignment in Figure 22 as reference, or s7) a polypeptide of at least two of s1 ), s2), s3), s4), s5) and s6); Further provided are recombinant microorganisms, wherein at least one polypeptide

having pamH activity, is provided by

t1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123 and/or 327, or

t2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 123, 124, 125, 126, 127, 128, 328, 329, 330, 331 , 332 and/or 333, or

t3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 122 and/or 326, or

t4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 122 and/or 326, or

a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 122 and/or 326 per primer, or

a polypeptide of t1 ) and/or t2) and comprising the amino acids D 16, N 1 17, H193, P1 19, H227 and N317 using the alignment in Figure 20 as reference, or

a polypeptide of at least two of t1), t2), t3), t4), t5) and t6).

The invention provides also for recombinant microorganism wherein

a) at least one polypeptide having pamC activity, is provided by

a1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 4 and/or 209, or

a2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, 5, 6, 7, 8, 9, 209,

210, 21 1 , 212, 213 and/or 214, or

a3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3 and/or 208 or

a4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 3 and/or 208, or

a5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 3 and/or 208 per primer, or

a6) a polypeptide of a1 ) and/or a2) and comprising the amino acids L29, D32,

S33, E44 and F47 using the alignment in Figure 3 as reference, or a7) a polypeptide of at least two of a1 ), a2), a3), a4), a5) and a6); b) at least one polypeptide having pamG activity, is provided by

b1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 1 1 and/or 216, or

b2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 , 12, 13, 14, 15, 16,

216, 217, 218, 219, 220 and/or 221 , or

b3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 10 and/or 215, or b4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 10 and/or 215, or

b5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 10 and/or 215 per primer, or

b6) a polypeptide of b1) and/or b2) and comprising the amino acids P19, A64,

C1 16, L139, D163, A167, H258, G287 and N288 using the alignment in Figure 4 as reference, or

b7) a polypeptide of at least two of b1 ), b2), b3), b4), b5) and b6);

c) at least one polypeptide having pamF activity, is provided by

c1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18 and/or 223, or

c2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 18, 19, 20, 21 , 22, 23,

223, 224, 225, 226, 227 and/or 228, or

c3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17 and/or 222, or

c4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 17 and/or 222, or

c5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 17 and/or 222 per primer, or

c6) a polypeptide of c1 ) and/or c2) and comprising the amino acids G16, A186, G255, G261 , G366 and H267 using the alignment in Figure 5 as reference, or

c7) a polypeptide of at least two of c1 ), c2), c3), c4), c5) and c6);

d) at least one polypeptide having pamA activity, is provided by

d1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 25 and/or 230, or

d2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, 26, 27, 28, 29, 30, 230, 231 , 232, 233, 234 and/or 235, or

d3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 24 and/or 229, or d4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 24 and/or 229, or

d5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 24 and/or 229 per primer, or

d6) a polypeptide of d1) and/or d2) and comprising the amino acids G21 ,

C191 , G258, G264, H333, G371 and H272 using the alignment in Figure 6 as reference, or

d7) a polypeptide of at least two of d1 ), d2), d3), d4), d5) and d6);

e) at least one polypeptide having pamB activity, is provided by

e1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 237, 349, 356, 363, 370 and/or 377, or

e2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 33, 34, 35, 36, 37, 237, 238, 239, 240, 241 , 242, 249, 350, 351 , 352, 353, 354, 356, 357, 358, 359, 360, 361 , 363, 364, 365, 366, 367, 368, 369, 371 , 372, 373, 374, 375, 377, 378, 379, 380, 381 and/or 382, or

e3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, or

e4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, or

e5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 31 , 236, 348, 355, 362, 369 and/or 376 per primer, or

e6) a polypeptide of e1) and/or e2) and comprising the amino acids T2, G3, P6, R22, A23, P59, L89, D218, G221 , N222, P260, P266, V270, G407, G409, E434, G436, G491 and at least two of G506, G508, G509 using the alignment in Figure 7 as reference, or

e7) a polypeptide of at least two of e1 ), e2), e3), e4), e5) and e6);

f) at least one polypeptide having pamD activity, is provided by

f1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39 and/or 244, or

f2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, 40, 41 , 42, 43, 44, 244, 245, 246, 247, 248 and/or 249, or

f3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38 and/or 243, or

f4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 38 and/or 243, or

f5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ I D

NO: 38 and/or 243 per primer, or

f6) a polypeptide of f1) and/or f2) and comprising the amino acids P21 , P101 ,

C121 , H246, G275 and G308 using the alignment in Figure 8 as reference, or

f7) a polypeptide of at least two of f1), f2), f3), f4), f5) and f6);

g) at least one polypeptide having pamE activity, is provided by

g1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46 and/or 251 , or

g2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 46, 47, 48, 49, 50, 51 , 251 , 252, 253, 254, 255 and/or 256, or

g3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45 and/or 250, or

g4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 45 and/or 250, or

g5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 45 and/or 250 per primer, or

g6) a polypeptide of g1) and/or g2) and comprising the amino acids P25, P103, C136, H270, G299 and G333 using the alignment in Figure 9 as reference, or

g7) a polypeptide of at least two of g 1 ), g2), g3), g4), g5) and g6);

h) at least one polypeptide having pamO activity, is provided by

hi ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, 258, 384, 391 , 398, 405 and/or 412, or

h2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, 54, 55, 56, 57, 58, 384, 385, 386, 387, 388, 389, 391 , 392, 393, 394, 395, 396, 398, 399, 400, 401 , 402, 403, 405, 406, 407, 408, 409, 410, 412, 413, 414, 415, 416 and/or 417, or

h3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , or

h4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ I D NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , or

h5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ I D NO: 52, 257, 383, 390, 397, 404 and/or 41 1 per primer, or

h6) a polypeptide of h i ) and/or h2) and comprising the amino acids G 1 1 , G 17,

D61 , Y164, K168, P 194 and at least two of G256, G257 and G258 using the alignment in Figure 10 as reference, or

h7) a polypeptide of at least two of h 1 ), h2), h3), h4), h5) and h6);

i) at least one polypeptide having pamK activity, is provided by

i 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 60 and/or 265, or

\2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 60, 61 , 62, 63, 64, 65, 265, 266, 267, 268, 269 and/or 270, or

13) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 59 and/or 264, or

14) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ I D

NO: 59 and/or 264, or

15) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ I D NO: 59 and/or 264 per primer, or

16) a polypeptide of i 1 ) and/or i2) and comprising the amino acids F33, C159, G314 and at least one of G346 and G348 using the alignment in Figure 1 1 as reference, or

17) a polypeptide of at least two of i 1 ) , i2), i3), i4), i5) and i6);

j) at least one polypeptide having pamJ activity, is provided by

j 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 67 and/or 272, or

j2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, 68, 69, 70, 71 , 72, 272, 273, 274, 275, 276 and/or 277, or

j3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66 and/or 271 , or

j4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 66 and/or 271 , or

j5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 66 and/or 271 per primer, or

j6) a polypeptide of j 1 ) and/or j2) and comprising the amino acids G14, G246,

G252, G256 and H257 using the alignment in Figure 12 as reference, or j7) a polypeptide of at least two of j 1 ) , j2), j3), j4), j5) and j6);

k) at least one polypeptide having pamM activity, is provided by

k1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 74 and/or 279, or

k2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, 75, 76, 77, 78, 79, 279, 280, 281 , 282, 283 and/or 284, or

k3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 73 and/or 278, or

k4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 73 and/or 278, or

k5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 73 and/or 278 per primer, or

k6) a polypeptide of k1 ) and/or k2) and comprising the amino acids G1 1 , V139,

G241 , G247, D291 , A319, Y384, K388, P414, G415, T419, G465, G473 and at least one of G472 and G473 using the alignment in Figure 13 as reference, or

k7) a polypeptide of at least two of k1 ), k2), k3), k4), k5) and k6);

I) at least one polypeptide having pamN activity, is provided by

11 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 and/or 286, or

I2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , 82, 83, 84, 85, 86, 286, 287, 288, 289, 290 and/or 291 , or

I3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 80 and/or 285, or

14) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 80 and/or 285, or

15) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 80 and/or 285 per primer, or

I6) a polypeptide of 11 ) and/or I2) and comprising the amino acids G16, G22,

D66, Y158, K162, P188 and G189 using the alignment in Figure 143 as reference, or

I7) a polypeptide of at least two of 11 ), I2), I3), I4), I5) and I6);

m) at least one polypeptide having pamL activity, is provided by

ml ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88 and/or 293, or

m2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, 89, 90, 91 , 92, 93, 293, 294, 295, 296, 297 and/or 298, or

m3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87 and/or 292, or

m4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 87 and/or 292, or

m5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 87 and/or 292 per primer, or

m6) a polypeptide of ml ) and/or m2) and comprising the amino acids L82, V93, T214, S215, G216, G219, K222, E352, G370, G394, Y410, T438, G439, D430, R445, G454, E463, P520, P530, G535, K536 and at least one of T217 and T218 using the alignment in Figure 15 as reference, or m7) a polypeptide of at least two of ml ), m2), m3), m4), m5) and m6);

n) at least one polypeptide having pamX activity, is provided by

n1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 95 and/or 300, or

n2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 95, 96, 97, 98, 99, 100, 300, 301 , 302, 303, 304 and/or 305, or

n3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 94 and/or 299, or

n4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ I D

NO: 94 and/or 299, or

n5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 94 and/or 299 per primer, or

n6) a polypeptide of n1) and/or n2) and comprising the amino acids D46, G57,

G1 13, H147, D241 , K271 , G276, T308 and at least one of G40 and G41 using the alignment in Figure 16 as reference, or

n7) a polypeptide of at least two of n 1 ), n2), n3), n4), n5) and n6);

o) at least one polypeptide having pamY activity, is provided by

01 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102 and/or 307, or

02) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102, 103, 104, 105,

106, 107, 307, 308, 309, 310, 31 1 and/or 312, or

03) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 101 and/or 306, or

o4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 101 and/or 306, or

05) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 101 and/or 306 per primer, or

06) a polypeptide of o1) and/or o2) and comprising the amino acids G67, G69, D86, and at least one of G 153 and G154, and at least one of G243 and G244 using the alignment in Figure 17 as reference, or

o7) a polypeptide of at least two of o1 ), o2), o3), o4), o5) and 06);

p) at least one polypeptide having pamS activity is provided by,

p1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 109 and/or 314, or

p2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, 1 10, 1 1 1 , 1 12, 1 13, 1 14, 314, 315, 316, 317, 318 and/or 319, or

p3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 108 and/or 313, or

p4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ I D

NO: 108 and/or 313, or

p5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 108 and/or 313 per primer, or

p6) a polypeptide of p1) and/or p2) and comprising the amino acids N41 , F76,

G79, D81 , G1 16, G122, D130, G148, G187, K221 and at least two of G 120

G121 and G122 using the alignment in Figure 18 as reference, or p7) a polypeptide of at least two of p1 ), p2), p3), p4), p5) and p6).

Preferable the recombinant microorganism comprises

a) at least one polypeptide having pamC activity, provided by

a1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4 and/or 209, or

a2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, 5, 6, 7, 8, 9, 209,

210, 21 1 , 212, 213 and/or 214, or

a3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3 and/or 208 or

a4) a polypeptide of at least two of a1 ), a2) and a3);

b) at least one polypeptide having pamG activity, provided by

b1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 1 1 and/or 216, or

b2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 , 12, 13, 14, 15, 16, 216, 217, 218, 219, 220 and/or 221 , or

b3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 10 and/or 215, or b4) a polypeptide of at least two of b1 ), b2) and b3;

c) at least one polypeptide having pamF activity, provided by

c1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18 and/or 223, or

c2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 18, 19, 20, 21 , 22, 23,

223, 224, 225, 226, 227 and/or 228, or

c3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17 and/or 222, or

c4) a polypeptide of at least two of c1 ), c2) and c3);

d) at least one polypeptide having pamA activity, provided by

d1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 25 and/or 230, or

d2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, 26, 27, 28, 29, 30,

230, 231 , 232, 233, 234 and/or 235, or

d3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 24 and/or 229, or

d4) a polypeptide of at least two of d 1 ), d2) and d3);

e) at least one polypeptide having pamB activity, provided by

e1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 32, 237, 349, 356, 363, 370 and/or 377, or

e2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 33, 34, 35, 36, 37, 237, 238, 239, 240, 241 , 242, 249, 350, 351 , 352, 353, 354, 356, 357, 358,

359, 360, 361 , 363, 364, 365, 366, 367, 368, 369, 371 , 372, 373, 374, 375,

377, 378, 379, 380, 381 and/or 382, or

e3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, or

e4) a polypeptide of at least two of e1 ), e2) and e3);

f) at least one polypeptide having pamD activity, provided by

f 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 39 and/or 244, or

f2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 39, 40, 41 , 42, 43, 44,

244, 245, 246, 247, 248 and/or 249, or

f3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 38 and/or 243, or

f4) a polypeptide of at least two of f 1 ), f2) and f3);

g) at least one polypeptide having pamE activity, provided by

g 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 46 and/or 251 , or

g2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ I D NO: 46, 47, 48, 49, 50, 51 ,

251 , 252, 253, 254, 255 and/or 256, or

g3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 45 and/or 250, or

g4) a polypeptide of at least two of g 1 ), g2) and g3);

h) at least one polypeptide having pamO activity, provided by

h i ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D

NO: 53, 258, 384, 391 , 398, 405 and/or 412, or

h2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 53, 54, 55, 56, 57, 58, 384, 385, 386, 387, 388, 389, 391 , 392, 393, 394, 395, 396, 398, 399, 400, 401 , 402, 403, 405, 406, 407, 408, 409, 410, 412, 413, 414, 415, 416 and/or 417, or

h3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , or

h4) a polypeptide of at least two of h 1 ), h2) and h3;

i) at least one polypeptide having pamK activity, provided by

i 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 60 and/or 265, or

\2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 60, 61 , 62, 63, 64, 65, 265, 266, 267, 268, 269 and/or 270, or

13) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 59 and/or 264, or

14) a polypeptide of at least two of i 1 ), i2) and i3); at least one polypeptide having pamJ activity, provided by

j1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 67 and/or 272, or

j2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, 68, 69, 70, 71 , 72,

272, 273, 274, 275, 276 and/or 277, or

j3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66 and/or 271 , or

j4) a polypeptide of at least two of j 1 ) , j2) and j3);

at least one polypeptide having pamM activity, provided by

k1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 74 and/or 279, or

k2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, 75, 76, 77, 78, 79,

279, 280, 281 , 282, 283 and/or 284, or

k3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 73 and/or 278, or

k4) a polypeptide of at least two of k1 ), k2) and k3);

at least one polypeptide having pamN activity, provided by

11 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 and/or 286, or

12) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , 82, 83, 84, 85, 86, 286, 287, 288, 289, 290 and/or 291 , or

13) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 80 and/or 285, or

14) a polypeptide of at least two of 11 ), I2) and I3);

at least one polypeptide having pamL activity, provided by

ml ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 88 and/or 293, or

m2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, 89, 90, 91 , 92, 93,

293, 294, 295, 296, 297 and/or 298, or

m3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87 and/or 292, or

m4) a polypeptide of at least two of ml ), m2) and m3);

at least one polypeptide having pamX activity, provided by

n1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 95 and/or 300, or

n2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 95, 96, 97, 98, 99, 100,

300, 301 , 302, 303, 304 and/or 305, or

n3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 94 and/or 299, or

n4) a polypeptide of at least two of n 1 ), n2) and n3)

at least one polypeptide having pamY activity, provided by

01 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102 and/or 307, or

02) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102, 103, 104, 105, 106, 107, 307, 308, 309, 310, 31 1 and/or 312, or

03) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 101 and/or 306, or

04) a polypeptide of at least two of o1 ), o2) and o3);

at least one polypeptide having pamS activity provided by,

p1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 109 and/or 314, or

p2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, 1 10, 1 1 1 , 1 12,

1 13, 1 14, 314, 315, 316, 317, 318 and/or 319, or

p3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 108 and/or 313, or

p4) a polypeptide of at least two of p1 ), p2) and p3).

Another illustrative example for a recombinant microorganism is a for recombinant

microorganism, wherein

a) at least one polypeptide having pamC activity, is provided by

a1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, a2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, 5, 6, 7, 8, and/or 9, or

a3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3, or

a4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 3, or a5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 3 per primer, or a6) a polypeptide of a1 ) and/or a2) and comprising the amino acids L29, D32, S33,

E44 and F47, or

a7) a polypeptide of at least two of a1 ), a2), a3), a4), a5) and a6); at least one polypeptide having pamG activity, is provided by

b1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 , or b2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%,

98%, 99%, or 100% identical to SEQ ID NO: 1 1 , 12, 13, 14, 15 and/or 16, or b3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 10, or

b4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 10, or b5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 10 per primer, or b6) a polypeptide of b1 ) and/or b2) and comprising the amino acids P19, A64, C1 16,

L139, D163, A167, H258, G287 and N288, or

b7) a polypeptide of at least two of b1), b2), b3), b4), b5) and b6); at least one polypeptide having pamF activity, is provided by

c1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18, or c2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%,

98%, 99%, or 100% identical to SEQ ID NO: 18, 19, 20, 21 , 22 and/or 23, or c3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17, or

c4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 17, or c5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 17 per primer, or c6) a polypeptide of c1 ) and/or c2) and comprising the amino acids G16, A186,

G255, G261 , G366 and H267, or

c7) a polypeptide of at least two of c1), c2), c3), c4), c5) and c6); d) at least one polypeptide having pamA activity, is provided by

d1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, or d2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, 26, 27, 28, 29 and/or 30, or d3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 24, or

d4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 24, or d5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 24 per primer, or d6) a polypeptide of d1 ) and/or d2) and comprising the amino acids G21 , A190, G258, G264, G371 and H272, or G21 , C191 , G258, G264, H333, G371 and

H272, preferably comprising the amino acids G21 , A190, C191 , G258, G264, H333, G371 and H272 or

d7) a polypeptide of at least two of d 1 ), d2), d3), d4), d5) and d6); e) at least one polypeptide having pamB activity, is provided by

e1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, or e2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 33, 34, 35, 36 and/or 37, or e3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31 , or

e4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 31 , or e5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 31 per primer, or e6) a polypeptide of e1 ) and/or e2) and comprising the amino acids T2, G3, P6, R22, A23, P59, L89, D218, G221 , N222, P260, P266, V270, G407, G409, E434, G436, G491 and at least two of G506, G508, G509, or

e7) a polypeptide of at least two of e1 ), e2), e3), e4), e5) and e6); f) at least one polypeptide having pamD activity, is provided by

f 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, or f2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, 40, 41 , 42, 43 and/or 44, or f3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38, or

f4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 38, or f5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 38 per primer, or f6) a polypeptide of f1) and/or f2) and comprising the amino acids P21 , P101 , C121 , H246, G275 and G308, or

f7) a polypeptide of at least two of f 1 ), f2), f3), f4), f5) and f6); g) at least one polypeptide having pamE activity, is provided by

g1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46, or g2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%,

98%, 99%, or 100% identical to SEQ ID NO: 46, 47, 48, 49, 50 and/or 51 , or g3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45, or

g4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 45, or g5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 45 per primer, or g6) a polypeptide of g1 ) and/or g2) and comprising the amino acids P25, P103,

C136, H270, G299 and G333, or

g7) a polypeptide of at least two of g 1 ), g2), g3), g4), g5) and g6); h) at least one polypeptide having pamO activity, is provided by

hi ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, or h2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, 54, 55, 56, 57 and/or 58, or h3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 52, or h4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 52, or h5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 52 per primer, or h6) a polypeptide of hi ) and/or h2) and comprising the amino acids G1 1 , G17, D61 , Y164, K168, P 194 and at least two of G256, G257 and G258, or

h7) a polypeptide of at least two of hi ), h2), h3), h4), h5) and h6); i) at least one polypeptide having pamK activity, is provided by

11 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, or

12) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, 61 , 62, 63, 64 and/or 65, or i3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 59, or

i4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 59, or i5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 59 per primer, or i6) a polypeptide of i 1 ) and/or i2) and comprising the amino acids F33, C159, G314 and at least one of G346 and G348, or

i7) a polypeptide of at least two of i1 ), i2), i3), i4), i5) and i6); j) at least one polypeptide having pamJ activity, is provided by

j1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, or j2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%,

98%, 99%, or 100% identical to SEQ ID NO: 67, 68, 69, 70, 71 and/or 72, or j3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66, or

j4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 66, or j5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 66 per primer, or j6) a polypeptide of j1 ) and/or j2) and comprising the amino acids G14, G246, G252,

G256 and H257, or

j7) a polypeptide of at least two of j 1 ) , j2), j3), j4), j5) and j6); at least one polypeptide having pamM activity, is provided by

k1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, or k2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%,

98%, 99%, or 100% identical to SEQ ID NO: 74, 75, 76, 77, 78 and/or 79, or k3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 73, or

k4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 73, or k5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 73 per primer, or k6) a polypeptide of k1 ) and/or k2) and comprising the amino acids G1 1 , V139,

G241 , G247, D291 , A319, Y384, K388, P414, G415, T419, G465, G473 and at least one of G472 and G473, or

k7) a polypeptide of at least two of k1), k2), k3), k4), k5) and k6); at least one polypeptide having pamN activity, is provided by

11 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , or

12) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , 82, 83, 84, 85 and/or 86, or

13) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 80, or

14) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 80, or

15) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 80 per primer, or

16) a polypeptide of 11 ) and/or I2) and comprising the amino acids G16, G22, D66, Y158, K162, P188 and G189, or

17) a polypeptide of at least two of 11 ), I2), I3), I4), I5) and I6); at least one polypeptide having pamL activity, is provided by

ml ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, or m2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%,

98%, 99%, or 100% identical to SEQ ID NO: 88, 89, 90, 91 , 92 and/or 93, or m3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87, or

m4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 87, or m5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 87 per primer, or m6) a polypeptide of ml ) and/or m2) and comprising the amino acids L82, V93,

T214, S215, G216, G219, K222, E352, G370, G394, Y410, T438, G439, D430, R445, G454, E463, P520, P530, G535, K536 and at least one of T217 and

T218, or

m7) a polypeptide of at least two of ml), m2), m3), m4), m5) and m6); n) at least one polypeptide having pamX activity, is provided by

n1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 95, or n2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%,

98%, 99%, or 100% identical to SEQ ID NO: 95, 96, 97, 98, 99 and/or 100, or n3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 94, or

n4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 94, or n5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 94 per primer, or n6) a polypeptide of n1 ) and/or n2) and comprising the amino acids D46, G57,

G1 13, H147, D241 , K271 , G276, T308 and at least one of G40 and G41 , or n7) a polypeptide of at least two of n1 ), n2), n3), n4), n5) and n6);

o) at least one polypeptide having pamY activity, is provided by

01 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102, or

02) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102, 103, 104, 105, 106 and/or

107, or

03) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 101 , or

o4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 101 , or o5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 101 per primer, or a polypeptide of o1 ) and/or o2) and comprising the amino acids G67, G69, D86, and at least one of G 153 and G154, and at least one of G243 and G244, or a polypeptide of at least two of o1 ), o2), o3), o4), o5) and 06); at least one polypeptide having pamS activity is provided by,

p1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, or

p2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, 1 10, 1 1 1 , 1 12, 1 13 and/or 1 14, or

p3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 108, or

p4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 108, or p5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 108 per primer, or p6) a polypeptide of p1) and/or p2) and comprising the amino acids N41 , F76, G79,

D81 , G1 16, G122, D130, G148, G187, K221 and at least two of G120 G121 and

G122, or

p7) a polypeptide of at least two of p1 ), p2), p3), p4), p5) and p6).

A person having skill in the art will understand that the polypeptides and polynucleotides provided herein for different embodiments of recombinant microorganisms, polynucleotides and methods, can be combined in various ways. Thus, the combination of features of a1 ) to p6) as described above as well as the combinations of features of q1 ) to t7) as described fourther below, as well as the different percent of sequence identities mentioned in these featueres can be combined in any possible manner. Further examples for such combinations are provided below. They represent preferred combinations, but should not be understood to limit the kind of possible combinations in any way.

Preferably the recombinant microorganisms have

a7) at least two of a1 ) and a6), a2) and a6), a3) and a6), a4) and a6), a5) and a6);

b7) at least two of b1 ) and b6), b2) and b6), b3) and b6), b4) and b6), b5) and b6);

c7) at least two of c1 ) and c6), c2) and c6), c3) and c6), c4) and c6), c5) and c6);

d7) at least two of d1 ) and d6), d2) and d6), d3) and d6), d4) and d6), d5) and d6);

e7) at least two of e1 ) and e6), e2) and e6), e3) and e6), e4) and e6), e5) and e6);

f7) at least two of f 1 ) and f6), f2) and f6), f3) and f6), f4) and f6), f5) and f6); g7) at leas- wo of g1 ) and g6), g2) and g6), g3) and g6), g4) and g6), g5) and g6);

h7) at leas- wo of hi ) and h6), h2) and h6), h3) and h6), h4) and h6), h5) and h6);

i7) at leas- wo of i1 ) and i6), i2) and i6), i3) and i6), i4) and i6), i5) and i6);

j7) at leas- wo of j1 ) and j6), j2) and j6), j3) and j6), j4) and j6), j5) and j6);

k7) at leas- wo of k1 ) and k6), k2) and k6), k3) and k6), k4) and k6), k5) and k6);

17) at leas- wo of 11 ) and I6), I2) and I6), I3) and I6), I4) and I6), I5) and I6);

m7) at leas- wo of ml ) and m6), m2) and m6), m3) and m6), m4) and m6), m5) and n7) at leas- wo of n1 ) and n6), n2) and n6), n3) and n6), n4) and n6), n5) and n6);

07) at leas- wo of o1 ) and 06), o2) and 06), o3) and 06), o4) and 06), o5) and 06);

p7) at leas- wo of p1 ) and p6), p2) and p6), p3) and p6), p4) and p6), p5) and p6).

Also preferred, the recombinant microorganisms have

a7) at least two of a1 ), a2) and a6);

b7) at least two of b1 ), b2), and b6);

ol) at least two of c1 ), c2), and c6);

61) at least two of dl), d2),) and d6);

el) at least two of e1), e2), and e6);

f7) at least two of fl), f2), and f6);

g7) at least two of g1), g2), and g6);

hi) at least two of hi), h2), and h6);

i7) at least two of i 1 ) , i2), and i6);

j7) at least two of j1), j2), and j6);

k7) at least two of k1 ), k2), and k6);

17) at least two of 11 ), I2), and I6);

m7) at least two of ml ), m2), and m6);

n7) at least two of n1), n2) and n6);

07) at least two of o1), o2), and 06);

P7) at least two of p1), p2), and p6),

wherein the % identity of the polypeptide to the respective SEQ ID NO: is at least 90%, at least 95%, at least 98%, or 100%.

Also preferred, the recombinant microorganisms have

37) at least two of a1 ), a3) and a6);

b7) at least two of b1 ), b3), and b6);

c7) at least two of c1 ), c3), and c6);

d7) at least two of d1 ), d3),) and d6);

Θ7) at least two of e1 ), e3), and e6);

f7) at least two of f1), f3), and f6);

g7) at least two of g1 ), g3), and g6);

h7) at least two of hi), h3), and h6);

i7) at least two of i 1 ) , i3), and i6);

j7) at least two of j 1 ) , j3), and j6); k7) at least two of k1), k3), and k6);

17) at least two of 11 ), I3), and I6);

m7) at least two of ml ), m3), and m6);

n7) at least two of n1 ), n3) and n6);

o7) at least two of o1 ), o3), and 06);

P7) at least two of p1 ), p3), and p6),

wherein the % identity of the polypeptide to the respective SEQ ID NO: is at least 90%, at least 95%, at least 98%, or 100%, and the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 90%, at least 95%, at least 98%, or 100%.

Also preferred, the recombinant microorganisms have

a7) at least two of a1 ), a3) and a6);

b7) at least two of b1 ), b3), and b6);

c7) at least two of c1 ), c3), and c6);

d7) at least two of d1 ), d3),) and d6);

Θ7) at least two of e1 ), e3), and e6);

f7) at least two of f1), f3), and f6);

g7) at least two of g1 ), g3), and g6);

h7) at least two of hi ), h3), and h6);

i7) at least two of i 1 ) , i3), and i6);

j7) at least two of j 1 ) , j3), and j6);

k7) at least two of k1), k3), and k6);

I7) at least two of 11 ), I3), and I6);

m7) at least two of ml ), m3), and m6);

n7) at least two of n1 ), n3) and n6);

07) at least two of o1 ), o3), and 06);

P7) at least two of p1 ), p3), and p6),

wherein the % identity of the polypeptide to the respective SEQ ID NO: at least 98%, or 100%, and the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 90%, at least 95%, at least 98%, or 100%.

Also preferred, the recombinant microorganisms have

a3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, b3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, c3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, d3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, e3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, f3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, g3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, h3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, i3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, j3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, k3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, I3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, m3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, n3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, o3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, and

p3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%. Preferably all of the % identities of a3) to p3) are at least 80%, are at least 90%, are at least 95%, are at least 98%, or 100%.

Preferably all of the % identities of a3) to p3) are at least 90%, are at least 95%, are at least 98%, or 100%. These recombinant microorganisms can be man-made mutants of pamamycin producing microorganisms, or microorganisms which comprise at least one recombinant expression cassette for at least one of the polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY or pamS activity. The recombinant microorganisms as described above may or may not be able to produce at least one pamamycin. Preferably they are able to produce at least one pamamycin, in particular if they are man-made mutants of pamamycin producing organisms. The

recombinant microorganisms are preferably used in methods which involve the production of pamamycin by such microorganisms.

Preferably, the recombinant microorganisms belong to the group of bacteria, preferably to the group of Pseudomonadaceae, Myxobacteria, Enterobacteriaceae, Bacillaceae, Brevibacteriaceae, Corynebacteriaceae, Streptomycetaceae.

The recombinant microorganisms may also belong to the group of yeasts or filamentous fungi, for example but not excluding others, Kluyveromyces, Saccharomyces,

Schizosaccharomyces, Yarrowia, Pichia, preferred species are: Kluyveromyces lactis, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Yarrowia lipolytica and Pichia stipites. Even more preferred, the recombinant microorganism is Saccharomyces cerevisiae.

Preferred bacteria belong to the genus Escherichia, Bacillus, Brevibacterium,

Corynebacterium, Saccharotrix, Kitasatospora, Nonomuraea, Pseudomonas, Myxococcus, Streptomyces or Amycolatopsis.

Preferred bacterial species which do not belong to the genus Streptomyces or

Amycolatopsis are Escherichia coli, Bacillus subtilis, Brevibacterium flavum, Brevibacterium lactofermentum, Brevibacterium divarecatum, Corynebacterium, particularly the species Corynebacterium glutamicum, Corynebacterium glutamicum, Corynebacterium

acetoglutamicum, Corynebacterium acetoacidophilum, Corynebacterium callunae,

Corynebacterium ammoniagenes, Corynebacterium thermoaminogenes, Corynebacterium melassecola and Corynebacterium effiziens, Saccharotrix espanaensis, Saccharotrix saharensis, Saccharotrix hoggarensis, Kitasatospora griseola, Kitasatospora setae, Nonomuraea jiangxensis, Nonomuraea flexuosa, Pseudomonas putida, Pseudomonas syringae, Myxococcus xanthus and Myxococcus stipitatus

Preferred bacteria of the group Streptomyces or Amycolatopsis are selected from the group consisting of: Actinomadura madurae, Actinomyces violaceochromogenes, Kitasatosporia parulosa, Streptomyces achromogenes, Streptomyces acidiscabies, Streptomyces albus, Streptomyces ambofaciens, Streptomyces antibioticus, Streptomyces aureofaciens, Streptomyces aurantiacus, Streptomyces avermitilis, Streptomyces capreolus,

Streptomyces carcinostaticus, Streptomyces cervinus, Streptomyces clavuligerus, Streptomyces coelicolor, Streptomyces coeruleorubidus, Streptomyces cyaneofuscatus, Streptomyces davawensis, Streptomyces diastaticus, Streptomyces fradiae,

Streptomyces fulvissimus, Streptomyces ghanaensis, Streptomyces griseus,

Streptomyces griseoflavus, Streptomyces griseoviridus, Streptomyces hirsutus,

Streptomyces hygroscopicus, Streptomyces kanamyceticus, Streptomyces lavendulae, Streptomyces lividans, Streptomyces loidensis, Streptomyces natalensis, Streptomyces noursei, Streptomyces nodosum, Streptomyces olivaceus, Streptomyces platensis, Streptomyces peuceticus, Streptomyces pristinaespiralis, Streptomyces rimosus, Streptomyces roseosporus, Streptomyces sviceus, Streptomyces spectabilis,

Streptomyces tauricus, Streptomyces tendae, Streptomyces thioluteus, Streptomyces toxytricini, Streptomyces venezuelae, Streptomyces violaceoniger, Streptomyces violaceoruber, Streptomyces violaceus, Streptomyces viridochromogenes, Streptomyces violaceoruber and Streptomyces zaomyceticus. In particular the Streptomyces species Streptomyces albus, Streptomyces auratus, Streptomyces avermitilis, Streptomyces coelicolor, Streptomyces clavuligerus,

Streptomyces fulvissimus, Streptomyces ghanaensis, Streptomyces griseoflavus, Streptomyces hygroscopicus, Streptomyces lividans, Streptomyces pristinaespiralis, Streptomyces roseosporus, Streptomyces roseosporus, Streptomyces sviceus

Streptomyces viridochromogenes, Streptomyces violaceoruber.

Preferred Streptomyces strains are: Streptomyces alboniger ATCC 12461, Streptomyces alboniger DSM 40043, Streptomyces alboniger DSM 40412, Streptomyces albus B24108, Streptomyces albus J 1074, Streptomyces albus DSM40313, Streptomyces avermitilis SUKA17 or Streptomyces avermitilis SUKA22, Streptomyces coelicolor DSM40233, Streptomyces kitasatoensis JCM5000, Streptomyces kitasatoensis JCM5001 Streptomyces lividans ATCC69444. In particular preferred are microorganisms which already have the capacity to produce pamamycin such as, but not excluding others: Streptomyces alboniger ATCC 12461 Streptomyces alboniger DSM 40043, Streptomyces aurantiacus ATCC 19822 and

Streptomyces kitasatoensis JCM5001.

A person skilled in the art will understand that the deposit number provided for the description of the strains are only examples and that the same strains may also be accessible under different deposit numbers at other strain collections.

Preferably the recombinant microorganisms comprise at least one expression cassette for each one of the polypeptides providing pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity, which has at least 80%, 82%, 84%, 86%, or 88% sequence identity to the respective polypeptide encoded by the respective polypeptide encoding region of SEQ ID NO: 1 and/or 2, wherein an individual expression cassette may encode only one polypeptide having one of the above mentioned activities, but may also have an operon like structure, so that it encodes more than one polypeptide having the same or different activities selected from the ones described above. More preferred, the recombinant microorganisms comprise at least one expression cassette for each one of the polypeptides providing pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity, which has at least 90%, 91 %, 92%, 93%, 94%, or 95% sequence identity to the respective polypeptide encoded by the respective polypeptide encoding region of SEQ ID NO: 1 and/or 2. More preferred, the recombinant microorganisms comprise at least one expression cassette for each one of the polypeptides providing pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity, which has at least 96%, 97%, 98%, 99%, or 100% sequence identity to the respective polypeptide encoded by the respective polypeptide encoding region of SEQ ID NO: 1 and/or 2. Preferably the recombinant microorganisms comprise at least one expression cassette for each one of the polypeptides providing pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity, which has at least 80%, 82%, 84%, 86%, or 88% sequence identity to the respective polypeptide encoding region of SEQ ID NO: 1 and/or 2. More preferred, the recombinant

microorganisms comprise at least one expression cassette for each one of the polypeptides providing pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity, which has at least 90%, 91 %, 92%, 93%, 94%, or 95% sequence identity to the respective polypeptide encoding region of SEQ ID NO: 1 and/or 2. More preferred, the recombinant microorganisms comprise at least one expression cassette for each one of the polypeptides providing pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity, which has at least 96%, 97%, 98%, 99%, or 100% sequence identity to the respective polypeptide encoding region of SEQ ID NO: 1 and/or 2.

One way to produce recombinant microorganisms of the invention is to transform

microorganisms with polynucleotides comprising expression cassettes for one or more of the polypeptides providing pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity. Accordingly, the invention comprises recombinant microorganisms comprising

i) at least one polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2,

ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2 and comprising at least one expression cassette for

a) at least one polypeptide having pamC activity, and

b) at least one polypeptide having pamG activity, and

c) at least one polypeptide having pamF activity, and

d) at least one polypeptide having pamA activity, and

e) at least one polypeptide having pamB activity, and

f) at least one polypeptide having pamD activity, and

g) at least one polypeptide having pamE activity, and

h) at least one polypeptide having pamO activity, and

i) at least one polypeptide having pamK activity, and

j) at least one polypeptide having pamJ activity, and

k) at least one polypeptide having pamM activity, and

I) at least one polypeptide having pamN activity, and

m) at least one polypeptide having pamL activity, and

n) at least one polypeptide having pamX activity, and

o) at least one polypeptide having pamY activity, and p) at least one polypeptide having pamS activity

iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments

comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of i) or ii), if the fragments are combined.

Preferably the polynucleotide of ii) differs mainly in the intergenic regions, promoters and/or terminators from the sequence of SEQ ID NO: 1 and/or 2. Hence, the polynucleotide of ii) may have at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, sequence identity to SEQ ID NO: 1 and/or 2, and comprises polypeptide encoding regions for the polypeptides providing pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity, which have at least 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the

polypeptides described by the respective amino acid sequences mentioned in Tables 4 to 19.

In particular having at least 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the polypeptides described by the respective amino acid sequence of SEQ ID NOs: 4, 1 1 , 18, 25, 32, 39, 46, 53, 60, 67, 74, 81 , 88, 95, 102 and 109. Preferably the are encoded by polynucleotides which have at least 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the polynucleotides described by the respective polynucleotide sequence of SEQ ID NOs: 3, 10, 17, 24, 31 , 38, 45, 52, 59, 66, 73, 80, 87, 94, 101 and 108.

The recombinant microorganisms, as described above, which have the capacity to produce pamamycin, including man-made mutants of naturally occurring pamamycin producing organisms, can be used in methods to produce pamamycin, preferably pamamycin 607 and/or pamamycin 621. The produced pamaycins can be hydrolysed in order to produce compounds of Formula V or Formula VI. Methods to hydrolyse pamamycin in order to produce compounds of the Formula V or Formula VI, as well as methods to purify and use compounds of the Formula V or Formula VI have been disclosed in DE4134168 and DE4316836. However, the methods disclosed in DE4134168 and DE4316836 should not be understood to be limiting. A person skilled in the art will be able to use alternative methods to hydrolyse pamamycin and to purify

compounds of the Formula V or Formula VI.

Furthermore, a person skilled in the art will also be able to use the information provided in Figure 2 to identify or construct microorganisms which have the capacitiy to produce one or more of the compounds S-chain, L-chain or K-chain.

Accordingly, the invention includes microorganisms having the capacitiy to produce compound S-chain, L-chain, K-chain or a combination of these, as well as methods to produce compound S-chain, L-chain and K-chain, using such microorganisms. A microorganism capable to produce the compound S-chain comprises:

I) at least one expression cassette encoding a polypepti de having pamC activity, and

II) at least one expression cassette encoding a polypepti de having pamA activity, and

III) at least one expression cassette encoding a polypepti de having pamB activity, and

IV) at least one expression cassette encoding a polypepti de having pamD activity, and

V) at least one expression cassette encoding a polypepti de having pamE activity, and

VI) at least one expression cassette encoding a polypepti de having pamO activity, and

VII) at least one expression cassette encoding a polypept de having pamM activity, and

VIII) at least one expression cassette encoding a polypepti de having pamN activity, and

IX) at least one expression cassette encoding a polypept de having pamS activity.

Preferably, the microorganism capable to produce the compound S-chain does not comprise one or more, or does not comprise any, of the expression cassettes selected from aa) an expression cassette encoding a polypepti de having pamG activity,

bb) an expression cassette encoding a polypepti de having pamF activity,

cc) an expression cassette encoding a polypepti de having pamK activity,

dd) an expression cassette encoding a polypepti de having pamJ activity,

ee) an expression cassette encoding a polypepti de having pamL activity,

ff) an expression cassette encoding a polypepti de having pamX activity, and

gg) an expression cassette encoding a polypepti de having pamY activity.

A microorganism capable to produce the compound K-chain comprises:

at least one expression cassette encoding a polypepti de having pamC activity, and at least one expression cassette encoding a polypepti de having pamA activity, and at least one expression cassette encoding a polypepti de having pamB activity, and at least one expression cassette encoding a polypepti de having pamD activity, and at least one expression cassette encoding a polypepti de having pamE activity, and at least one expression cassette encoding a polypepti de having pamO activity, and at least one expression cassette encoding a polypepti de having pamM activity, and at least one expression cassette encoding a polypepti de having pamN activity, and at least one expression cassette encoding a polypept de having pamS activity, and at least one expression cassette encoding a polypepti de having pamG activity, and at least one expression cassette encoding a polypepti de having pamF activity.

Preferably, the microorganism capable to produce the compound K-chain does not comprise one or more, preferably does not comprise any, of the expression cassettes selected from

aa) an expression cassette encoding a polypeptide having pamK activity,

bb) an expression cassette encoding a polypeptide having pamJ activity,

cc) an expression cassette encoding a polypeptide having pamL activity,

dd) an expression cassette encoding a polypeptide having pamX activity, and ee) an expression cassette encoding a polypeptide having pamY activity.

Preferably, the microorganism capable to produce the compound K-chain does not comprise

an expression cassette encoding a polypeptide having pamX activity, and/or

an expression cassette encoding a polypeptide having pamY activity,

A microorganism capable to produce the compound L-chain comprises:

1) at least one expression cassette encoding a polypeptide having pamC activity, and

II) at least one expression cassette encoding a polypeptide having pamA activity, and ill) at least one expression cassette encoding a polypeptide having pamB activity, and iv) at least one expression cassette encoding a polypeptide having pamD activity, and

V) at least one expression cassette encoding a polypeptide having pamE activity, and

VI) at least one expression cassette encoding a polypeptide having pamO activity, and

VII) at least one expression cassette encoding a polypeptide having pamM activity, and

VIII) at least one expression cassette encoding a polypeptide having pamN activity, and

IX) at least one expression cassette encoding a polypeptide having pamS activity, and

X) at least one expression cassette encoding a polypeptide having pamG activity, and

XI) at least one expression cassette encoding a polypeptide having pamF activity, and

XII) at least one expression cassette encoding a polypeptide having pamX activity, and

XIII) at least one expression cassette encoding a polypeptide having pamY activity.

Preferably, the microorganism capable to produce the compound L-chain does not comprise one or more, preferably does not comprise any, of the expression cassettes selected from

aa) at least one expression cassette encoding a polypeptide having pamK activity, bb) at least one expression cassette encoding a polypeptide having pamJ activity, and cc) at least one expression cassette encoding a polypeptide having pamL activity. Microorganisms capable to produce at least one of the pamamycins described above, e.g. pamamycin 607 or 621 may be used to produce microorganisms for the production of least one of the compounds S-chain, K-chain and L-chain, by reducing the expression or the activity of one or more polypeptides having an activity which is used to produce pamamycin, but not being necessary to produce compound S-chain, K-chain and/or L-chain.

Accordingly, the microorganisms capable to produce compound S-chain may have a reduced activtiy of at least one of the expression cassettes selected from

aa) an expression cassette encoding a polypeptide having pamG activity,

bb) an expression cassette encoding a polypeptide having pamF activity,

cc) an expression cassette encoding a polypeptide having pamK activity,

dd) an expression cassette encoding a polypeptide having pamJ activity,

ee) an expression cassette encoding a polypeptide having pamL activity,

ff) an expression cassette encoding a polypeptide having pamX activity, and gg) an expression cassette encoding a polypeptide having pamY activity.

The microorganisms capable to produce compound K-chain may have a reduced activtiy of at least one of the expression cassettes selected from

aa) an expression cassette encoding a polypeptide having pamK activity, and

bb) an expression cassette encoding a polypeptide having pamJ activity, and

cc) an expression cassette encoding a polypeptide having pamL activity, and

dd) an expression cassette encoding a polypeptide having pamX activity, and

ee) an expression cassette encoding a polypeptide having pamY activity, and

The microorganisms capable to produce compound L-chain may have a reduced activtiy of at least one of the expression cassettes selected from

aa) at least one expression cassette encoding a polypeptide having pamK activity, bb) at least one expression cassette encoding a polypeptide having pamJ activity, and cc) at least one expression cassette encoding a polypeptide having pamL activity.

A reduced activtiy of at least one of the expression cassettes mentioned above can be achieved by reducing the expression level of the respective expression cassette, e.g. by the use of a less active promoter or less efficient terminator sequence, or by expressing a polypeptide having a reduced pamG, pamF, pamK, pamJ, pamL or pamX activity, or expressing a less stable polypeptide having pamG, pamF, pamK, pamJ, pamL or pamX activity, or by using a combination of a less active promoter, less efficient terminator, less active polypeptide or less stable polypeptide. The activity of the respective promoter, the efficiency of the terminator, the activity of the polypeptide and the stability of the polypeptide are compared to the respective promoter, terminator or polypeptide present in the wildtype microorganism.

The expression cassettes described above for the microorganisms capable to produce at least one of the compounds S-chain, K-chain and L-chain preferably encode polypeptides which can be described by the sequences defined above for the polypeptides having pamC, pamG, pamG, pamA, pamB, pamD, pamE, pamO, pamK, pamK, pamM, pamN, pamL, pamX, pamY or pamS activity. For example:

for pamC activity the sequences described in Table 4, Table 4a, Figure 3 and a1 to a7, for pamG activity the sequences described in Table 5, Table 5a, Figure 4 and b1 to b7, for pamF activity the sequences described in Table 6, Table 6a, Figure 5 and c1 to c7, for pamA activity the sequences described in Table 7, Table 7a, Figure 6 and d1 to d7, for pamB activity the sequences described in Table 8, Table 8a to 8g, Figure 7 and e1 to e7,

for pamD activity the sequences described in Table 9, Table 9a, Figure 8 and f1 to f7, for pamE activity the sequences described in Table 10, Table 10a, Figure 9 and g1 to g7, for pamO activity the sequences described in Table 1 1 , Table 1 1a to 1 1g, Figure 10 and hi to h7, for pamK activity the sequences described in Table 12, Table 12a, Figure 1 1 and i1 to i7, for pamJ activity the sequences described in Table 13, Table 13a, Figure 12 and j 1 to j7, for pamM activity the sequences described in Table 14, Table 14a, Figure 13 and k1 to k7, for pamN activity the sequences described in Table 15, Table 15a, Figure 14 and 11 to I7, for pamL activity the sequences described in Table 16, Table 16a, Figure 15 and ml to m7, for pamX activity the sequences described in Table 17, Table 17a, Figure 16 and n1 to n7, for pamY activity the sequences described in Table 18, Table 18a, Figure 17 and o1 to o7, for pamS activity the sequences described in Table 19, Table 19a, Figure 18 and p1 to p7,

Methods to produce compound S-chain, L-chain and K-chain, comprise the following steps: i) cultivating a recombinant microorganism capable to produce at least one of compounds S-chain, K-chain and L-chain under conditions which allow for the production at least one of compounds S-chain, K-chain and L-chain by said recombinant microorganism;

ii) obtaining at least one of compounds S-chain, K-chain and L-chain, preferably obtaining compound L-chain.

The microorganisms capable to produce at least one of the compounds S-chain, K-chain and L-chain preferably comprise at least one recombinant expression cassette which encodes a polypeptide which can be described by the sequences defined above for the polypeptides having pamC, pamG, pamG, pamA, pamB, pamD, pamE, pamO, pamK, pamK, pamM, pamN, pamL, pamX, pamY or pamS activity. The recombinant microorganisms, as described above for the production of pamamycin , S- chain, L-chain or K-chain, preferably comprise at least one expression cassette for q) a polypeptide having pamR1 activity, or

r) a polypeptide having pamR2 activity, or

s) a polypeptide having pamW activity, or

t) a polypeptide having pamH activity, or

u) a polypeptide q), r) s) and t) and at least one expression cassette for a

polypeptide of q), r) s) and t), which encodes a different polypeptide than the first expression cassette. Preferably, they comprise at least one expression cassette for a polypeptide for each one of pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY and pamS activity, e.g. by comprising

i) several independent expression cassettes located in the genome, or

ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2 and comprising at least one expression cassette for at least one polypept de having pamC activity, and

at least one polypept de having pamG activity, and

at least one polypept de having pamF activity, and

at least one polypept de having pamA activity, and

at least one polypept de having pamB activity, and

at least one polypept de having pamD activity, and

at least one polypept de having pamE activity, and

at least one polypept de having pamO activity, and

at least one polypept de having pamK activity, and

at least one polypept de having pamJ activity, and

at least one polypept de having pamM activity, and

at least one polypept de having pamN activity, and

at least one polypept de having pamL activity, and

at least one polypept de having pamX activity, and

at least one polypept de having pamY activity, and

at least one polypept de having pamS activity

iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments

comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of i) or ii), if the fragments are combined, and comprise at least one expression cassette for

q) a polypeptide having pamR1 activity, or

r) a polypeptide having pamR2 activity, or

s) a polypeptide having pamW activity, or

t) a polypeptide having pamH activity.

The polypeptides having pamR1 , pamR2, pamW or pamH activity are preferably provided by endogenous expression cassettes or on recombinant expression cassettes.

The recombinant microorganism preferably comprise expression cassettes, wherein the polypeptide having pamR1 activity, is provided by

q1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 1 16 and/or 321 , or

q2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17, 1 18, 1 19,

120, 121 , 321 , 322, 323, 324, 325 and/or 326, or

q3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15 and/or 320, or

q4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 1 15 and/or 320, or

a polypeptide expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 and/or 320 per primer, or

a polypeptide of q1) and/or q2) and comprising the amino acids K105, K190 and at least one of E153 and E154 using the alignment in Figure 19 as reference, or

a polypeptide of at least two of q1 ), q2), q3), q4), q5) and q6), more preferred, the polypeptide having pamR1 activity, is provided by

q1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 1 16 and/or 321 , or

q2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17, 1 18, 1 19,

120, 121 , 321 , 322, 323, 324, 325 and/or 326, or

q3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15 and/or 320, or

q7) a polypeptide of at least two of q 1 ), q2) and q3).

The recombinant microorganism preferably comprise expression cassettes, wherein the polypeptide having pamR2 activity, is provided by

r1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 130 and/or 335, or

r2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, 131 , 132, 133,

134, 135, 335, 336, 337, 338, 339 and/or 340, or

r3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 129 and/or 334, or

r4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 129 and/or 334, or

r5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 129 and/or 334 per primer, or

r6) a polypeptide of r1 ) and/or r2) and comprising the amino acids E3, W19,

A59, Y70, G169, P207, F226, L230, G236 at least one of A40 and A41 and at least one of G48 and G49 and at least one of L79 and L80 usi

alignment in Figure 21 as reference, or

a polypeptide of at least two of r1 ), r2), r3), r4), r5) and r6); more preferred, the polypeptide having pamR2 activity, is provided by

r1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 130 and/or 335, or

r2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, 131 , 132, 133,

134, 135, 335, 336, 337, 338, 339 and/or 340, or

r3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 129 and/or 334, or

r4) a polypeptide of at least two of r1 ), r2) and r3).

The recombinant microorganism preferably comprise expression cassettes, wherein the polypeptide having pamW activity, is provided by

s1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 137 and/or 342, or

s2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, 138, 139, 140,

141 , 142, 342, 343, 344, 345, 346 and/or 347, or

s3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 136 and/or 341 , or

s4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 136 and/or 341 , or

s5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 136 and/or 341 per primer, or

s6) a polypeptide of s1 ) and/or s2) and comprising the amino acids D92, G95 and G131 using the alignment in Figure 22 as reference, or s7) a polypeptide of at least two of s1 ), s2), s3), s4), s5) and s6); more preferred, the polypeptide having pamW activity, is provided by

s1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137 and/or 342, or s2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, 138, 139, 140, 141 , 142, 342, 343, 344, 345, 346 and/or 347, or

s3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 136 and/or 341 , or

s4) a polypeptide of at least two of s1 ), s2) and s3).

The recombinant microorganism preferably comprise expression cassettes, wherein the polypeptide having pamH activity, is provided by

t1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123 and/or 327, or

t2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123, 124, 125, 126,

127, 128, 328, 329, 330, 331 , 332 and/or 333, or

t3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 122 and/or 326, or

t4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 122 and/or 326, or

t5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID

NO: 122 and/or 326 per primer, or

t6) a polypeptide of t1) and/or t2) and comprising the amino acids D16, N1 17, H193, P1 19, H227 and N317 using the alignment in Figure 20 as reference, or

t7) a polypeptide of at least two of t1 ), t2), t3), t4), t5) and t6); more preferred, the polypeptide having pamH activity, is provided by

t1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123 and/or 327, or

t2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123, 124, 125, 126, 127, 128, 328, 329, 330, 331 , 332 and/or 333, or

t3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 122 and/or 326, or

t4) a polypeptide of at least two of t1 ), t2) and t3). Also preferred are expression cassettes providing for

q) a polypeptide having pamR1 activity

q1 ) which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, or

q2) which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or

100% identical to SEQ ID NO: 1 16, 1 17, 1 18, 1 19, 120 and/or 121 , or q3) which is expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15, or

q4) which is expressed from a polynucleotide which hybridises under medium

stringency hybridisation conditions, to a complement of SEQ ID NO: 1 15, or q5) which is expressed from a polynucleotide which is obtainable via PCR on

genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 per primer, or q6) by being a polypeptide of q1 ) and/or q2) and comprising the amino acids K105,

K190 and at least one of E153 and E154, or

q7) by being a polypeptide of at least two of q 1 ), q2), q3), q4), q5) and q6); r) polypeptide having pamR2 activity

r1 ) which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%,

96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, or

r2) which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, 131 , 132, 133, 134 and/or 135, or r3) which is expressed from a polynucleotide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 129, or

r4) which is expressed from a polynucleotide which hybridises under medium

stringency hybridisation conditions, to a complement of SEQ ID NO: 129, or r5) which is expressed from a polynucleotide which is obtainable via PCR on

genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 129 per primer, or r6) by being a a polypeptide of r1 ) and/or r2) and comprising the amino acids E3, W19, A59, Y70, G169, P207, F226, L230, G236 at least one of A40 and A41 and at least one of G48 and G49 and at least one of L79 and L80, or r7) by being a polypeptide of at least two of r1), r2), r3), r4), r5) and r6); s) polypeptide having pamW activity,

s1 ) which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, or

s2) which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, 138, 139, 140, 141 and 142, or which is expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 136, or

which is expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 136, or which is expressed from a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pai of at least 15 consecutive polynucleotides of SEQ ID NO: 136 per primer, or by being a polypeptide of s1 ) and/or s2) and comprising the amino acids D92, G95 and G131 , or

by being a polypeptide of at least two of s1), s2), s3), s4), s5) and s6); t) polypeptide having pamH activity,

t1 ) which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123, or

t2) which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or

100% identical to SEQ ID NO: 123, 124, 125, 126, 127 and/or 128, or t3) which is expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 122, or

t4) which is expressed from a polynucleotide which hybridises under medium

stringency hybridisation conditions, to a complement of SEQ ID NO: 122, or t5) which is expressed from a polynucleotide which is obtainable via PCR on

genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 122 per primer, or t6) by being a polypeptide of t1) and/or t2) and comprising the amino acids D16,

N1 17, H 193, P1 19, H227 and N317, or

t7) by being a polypeptide of at least two of t1 ), t2), t3), t4), t5) and t6). Preferably the recombinant microorganisms have

a7) at least two of a1 ) and a6), a2) and a6), a3) and a6), a4) and a6), a5) and a6); b7) at least two of b1 ) and b6), b2) and b6), b3) and b6), b4) and b6), b5) and b6); c7) at least two of c1 ) and c6), c2) and c6), c3) and c6), c4) and c6), c5) and c6);

d7) at least two of d 1 ) and d6), d2) and d6), d3) and d6), d4) and d6), d5) and d6); e7) at least two of e1 ) and e6), e2) and e6), e3) and e6), e4) and e6), e5) and e6); f7) at least two of f1 ) and f6), f2) and f6), f3) and f6), f4) and f6), f5) and f6);

g7) at least two of g 1 ) and g6), g2) and g6), g3) and g6), g4) and g6), g5) and g6); h7) at least two of hi ) and h6), h2) and h6), h3) and h6), h4) and h6), h5) and h6); i7) at least two of i1 ) and i6), i2) and i6), i3) and i6), i4) and i6), i5) and i6);

j7) at least two of j1 ) and j6), j2) and j6), j3) and j6), j4) and j6), j5) and j6);

k7) at least two of k1 ) and k6), k2) and k6), k3) and k6), k4) and k6), k5) and k6);

I7) at least two of 11 ) and I6), I2) and I6), I3) and I6), I4) and I6), I5) and I6); m7) at least two

n7) at least two

07) at least two

P7) at least two

q7) at least two

r7) at least two

s7) at least two

t7) at least two

Also preferred, the

a7) at least two

b7) at least two

c7) at least two

d7) at least two

Θ7) at least two

f7) at least two

g7) at least two

h7) at least two

i7) at least two

j7) at least two

k7) at least two

I7) at least two

m7) at least two

n7) at least two

07) at least two

P7) at least two

q7) at least two

r7) at least two

s7) at least two

t7) at least two

wherein the % identity of the polypeptide to the respective SEQ ID NO: is at least at least 90%, at least 95%, at least 98%, or 100%.

Also preferred, the recombinant microorganisms have

a7) at least two of a 1 ), a3) and a6);

b7) at least two of b1 ), b3), and b6);

c7) at least two of c1 ), c3), and c6);

d7) at least two of d1 ), d3),) and d6);

e7) at least two of e1 ), e3), and e6);

f7) at least two of f1), f3), and f6);

g7) at least two of g1 ), g3), and g6);

h7) at least two of hi ), h3), and h6); i7) at least two of i 1 ) , i3), and i6);

j7) at least two of j 1 ) , j3), and j6);

k7) at least two of k1), k3), and k6);

17) at least two of 11 ), I3), and I6);

m7) at least two of ml ), m3), and m6);

n7) at least two of n1 ), n3) and n6);

07) at least two of o1 ), o3), and 06);

P7) at least two of p1 ), p3), and p6),

q7) at least two of q1), q2) and q6;

r7) at least two of r1 ), r2) and r6);

s7) at least two of s1 ), s2) and s6);

t7) at least two of t1), t2) and t6,

wherein the % identity of the polypeptide to the respective SEQ ID NO: is at least 90%, at least 95%, at least 98%, or 100%, and the % identity of the polypeptide encoding

polynucleotide to the respective SEQ ID NO: is at least 90%, at least 95%, at least 98%, or 100%.

Also preferred, the recombinant microorganisms have

a7) at least two of a1 ), a3) and a6);

b7) at least two of b1 ), b3), and b6);

c7) at least two of c1 ), c3), and c6);

d7) at least two of d1 ), d3),) and d6);

Θ7) at least two of e1 ), e3), and e6);

f7) at least two of f1), f3), and f6);

g7) at least two of g1), g3), and g6);

h7) at least two of hi ), h3), and h6);

i7) at least two of i 1 ) , i3), and i6);

j7) at least two of j 1 ) , j3), and j6);

k7) at least two of k1), k3), and k6);

I7) at least two of 11 ), I3), and I6);

m7) at least two of ml ), m3), and m6);

n7) at least two of n1 ), n3) and n6);

07) at least two of o1 ), o3), and 06);

P7) at least two of p1 ), p3), and p6),

q7) at least two of q1 ), q3) and q6;

r7) at least two of r1 ), r3) and r6);

s7) at least two of s1 ), s3) and s6);

t7) at least two of t1), t3) and t6).

wherein the % identity of the polypeptide to the respective SEQ ID NO: at least 98%, or 100%, and the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 90%, at least 95%, at least 98%, or 100%. Also preferred, the recombinant microorganisms have

a3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, b3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, c3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, d3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, e3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, f3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, g3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, h3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, i3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, j3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, k3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, I3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, m3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, n3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, o3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, p3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, q3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, r3) wherein the % identity of the polypeptide encoding polynucleotide to the respective

SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%, s3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100% and

t3) wherein the % identity of the polypeptide encoding polynucleotide to the respective SEQ ID NO: is at least 80%, is at least 90%, is at least 95%, at least 98%, or 100%. Preferably all of the % identities of a3) to t3) are at least 80%, are at least 90%, are at least 95%, are at least 98%, or 100%.

Preferably all of the % identities of a3) to t3) are at least 90%, are at least 95%, are at least 98%, or 100%.

The recombinant microorganisms described above may comprise

i) at least one polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID

NO: 1 ,

ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and comprising at least one expression cassette for

at least one polypept de having pamC activity, and

at least one polypept de having pamG activity, and

at least one polypept de having pamF activity, and

at least one polypept de having pamA activity, and

at least one polypept de having pamB activity, and

at least one polypept de having pamD activity, and

at least one polypept de having pamE activity, and

at least one polypept de having pamO activity, and

at least one polypept de having pamK activity, and

at least one polypept de having pamJ activity, and

at least one polypept de having pamM activity, and

at least one polypept de having pamN activity, and

at least one polypept de having pamL activity, and

at least one polypept de having pamX activity, and

at least one polypept de having pamY activity, and

at least one polypept de having pamS activity, and

at least one polypept de having pamR1 activity, and

at least one polypept de having pamR2 activity, and

at least one polypept de having pamW activity, and

at least one polypept de having pamH activity, and

iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments

comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of i) or ii), if the fragments are combined.

Preferably the polynucleotide of ii) differs mainly in the intergenic regions, promoters and/or terminators from the sequence of SEQ ID NO: 2. Hence, the polynucleotide of ii) may have at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, sequence identity to SEQ ID NO: 2, and comprises polypeptide encoding regions for the polypeptides providing pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS, pamR1 , pamR2, pamW and pamH activity, which have at least 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the polypeptides described by the respective amino acid sequences mentioned in Tables 4 to 23.

In particular having at least 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the polypeptides described by the respective amino acid sequence of SEQ ID NOs: 4, 1 1 , 18, 25, 32, 39, 46, 53, 60, 67, 74, 81 , 88, 95, 102, 109, 1 16, 123, 130, and 137. Preferably the are encoded by polynucleotides which have at least 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the polynucleotides described by the respective polynucleotide sequence of SEQ ID NOs: 3, 10, 17, 24, 31 , 38, 45, 52, 59, 66, 73, 80, 87, 94, 101 , 108 1 15, 122, 129 and 139.

In one embodiment the recombinant microorganisms comprise polypeptides for pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS and or pamR1 activity, preferably for pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS, pamR1 and pamW activity.

In further embodiments the recombinant microorganisms comprise polypeptides for pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS and or pamR1 activity, but no or a reduced activity of pamR1 and/or pamH, preferably for pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS, pamR1 and pamW activity, but no or a reduced activity of pamR1 and/or pamH.

The recombinant microorganisms as described above may or may not be able to produce at least one pamamycin. Preferably they are able to produce at least one pamamycin, in particular if they are man-made mutants of pamamycin producing organisms. The recombinant microorganisms are preferably used in methods which involve the production of pamamycin by such microorganisms.

The recombinant microorganisms do preferably belong to a species as described above, more preferred they belong to the group of Streptomyces, e.g. the Streptomyces species described above, in particular to the pamamicin producing species described above. A particular preferred part of the invention are recombinant microorganisms comprising i) at least one polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2,

ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2 and comprising at least one expression cassette for a) at least one polypeptide having pamC activity, and

b) at least one polypeptide having pamG activity, and

c) at least one polypeptide having pamF activity, and

d) at least one polypeptide having pamA activity, and

e) at least one polypeptide having pamB activity, and

f) at least one polypeptide having pamD activity, and

g) at least one polypeptide having pamE activity, and

h) at least one polypeptide having pamO activity, and

i) at least one polypeptide having pamK activity, and

j) at least one polypeptide having pamJ activity, and

k) at least one polypeptide having pamM activity, and

1) at least one polypeptide having pamN activity, and

m) at least one polypeptide having pamL activity, and

n) at least one polypeptide having pamX activity, and

o) at least one polypeptide having pamY activity, and

P) at least one polypeptide having pamS activity

two or more fragments of the polynucleotides of i) or ii) wherein the fragments comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of i) or ii), if the fragments are combined

and comprising a recombinant expression cassette for a polypeptide having pamR1 activity is provided by

q1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, or

q2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17, 1 18, 1 19, 120 and/or 121 , or

q3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15, or

q4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID

NO: 1 15, or

q5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 per primer, or

q6) a polypeptide of q 1 ) and/or q2) and comprising the amino acids K105, K190 and at least one of E 153 and E 154, or

q7) a polypeptide of at least two of q 1 ), q2), q3), q4), q5) and q6). Preferred embodiments define the polypeptides of a) to p) according to the criteria of a1 ) to p7) as described above, including preferred combinations and sequence identity regions of these criteria as described above. Preferably the polypeptide comprising pamR1 activity is expressed under control of a promoter which does not comprise a polynucleotide sequence of SEQ ID NO: 145, and preferably provides for a constitutive or inducible expression in the recombinant

microorganism.

In further embodiments the recombinant microorganism comprises also a polypeptide having pamW activity, provided by

s1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, or

s2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, 138, 139, 140, 141 and 142, or

s3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 136, or

s4) a polypeptide expressed from a polynucleotide which hybridises under medium

stringency hybridisation conditions, to a complement of SEQ ID NO: 136, or s5) a polypeptide expressed from a polynucleotide which is obtainable via PCR on

genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 136 per primer, or

s6) a polypeptide of s1 ) and/or s2) and comprising the amino acids D92, G95 and G131 , or

s7) a polypeptide of at least two of s1), s2), s3), s4), s5) and s6);

Preferred embodiments define the polypeptides of a) to p), or the polypeptides of a) to s) as applicable according to the criteria of a1 ) to p7) or al ) to s7), respectively, as described above, including preferred combinations and sequence identity regions of these criteria as described above.

Even more preferred, are microoranisms comprising expression cassettes for polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS activity, or comprising expression cassettes for polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS pamR1 and/or pamW activity, or comprising expression cassettes for polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS pamR1 and pamW, wherein the polynucleotides encoding the respective polypeptides are obtainable from Streptomyces alboniger ATCC 12461 Streptomyces alboniger DSM 40043, Streptomyces aurantiacus ATCC 19822 and/or Streptomyces kitasatoensis JCM5001. The sequence information of polynucleotides isolated from the natural sources described above can be used to isolate homologous polynucleotides of the genes, promoter and terminator sequences comprised by SEQ ID NO: 1 and/or 2, as well as homologous versions of SEQ ID NO: 1 and/or 2. Further variants of the disclosed polynucleotides can be constructed, e.g. by adapting the codon usage of polypeptide encoding nucleic acid sequences to the codon usage of a preferred species of microorganism, or by exchanging promoter regions and/or terminator regions or both of an expression cassette in order to adapt the expression of an encoded polynucleotide to a preferred species of microorganism or culture conditions.

Polynucleotide variants encoding polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS, pamR1 , pamH, pamR2 or pamW activity and having a sequence identity of at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% to the polynucleotide sequences encoding the respective polpeptide on SEQ ID NO: 1 and/or 2 can be readily constructed by adapting the codon usage to the codon usage of a specific microorganism of interst and/or by changes to a encoded poypeptide variant having the respective activity, for example but not excluding others, by encoding one of the polypeptide variants mentioned in Tables 4 to 23.

Further variants of the polynucleotides of the invention can be created by adding, deleting one or more polynucleotides from a polynucleotide, e.g. by shortening spacer regions between expression cassettes, or deleting one or more codons of polypeptide encoding regions or complete functional elements of the polynucleotides, like complete, promoter, terminator or polypeptide encoding regions or complete expression cassettes. Alternatively, or in addition thereto, it is possible to create variants of the encoded polypeptide

sequences, e.g. by introducing conserved amino acid substitutions or by adding or deleting 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10 or more codons in order to enlarge or shorten the encoded polypeptides, or to create polypeptide fusions. Preferred polypeptide fusion comprise polypeptides for monitoring expression (e.g., green, yellow, blue or red fluorescent proteins, alkaline phosphatase and the like) or so called " tags" which may serve as a detectable marker or as an auxiliary measure for purification purposes. Tags for the different purposes are well known in the art and comprise FLAG-tags, 6-histidine-tags, MYC-tags and the like. The variant nucleic acid sequence shall still encode a polypeptide being involved in Pamamycin synthesis. These polynucleotides and polypeptides can be used to construct recombinant

microorganisms, in particular recombinant microorganisms for the production of at least one pamamycin, as described above, but can also be used for other purposes, e.g. but not excluding others, to overproduce polypeptides for in-vitro applications, protein interaction assays or to develop antibodies against the polypeptides.

Accordingly, the invention comprises also microorganisms which comprise at least one expression cassette as described in a1 ) to t7) below a) an expression cassette comprising

a1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4 and/or 209, preferably to SEQ ID NO: 4, or

a2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, 5, 6, 7, 8, 9, 209, 210, 21 1 , 212, 213 and/or 214, preferably to SEQ ID NO: 4, 5, 6, 7, 8, and/or 9, or

a3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3 and/or 208, preferably to SEQ ID NO: 3, or

a4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 3 and/or 208, preferably of SEQ ID NO: 3, or

a5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 3 and/or 208, preferably of SEQ ID NO: 3 per primer, or

a6) a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a pamamycin producing organism, or

a7) a polynucleotide of at least two of a1 ), a2), a3), a4), a5) and a6);

b) an expression cassette comprising

b1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or

100% identical to SEQ ID NO: 1 1 and/or 216, preferably to SEQ ID NO: 1 1 , or

b2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 , 12, 13, 14, 15, 16, 216, 217, 218, 219, 220 and/or 221 , preferably to

SEQ ID NO: 1 1 , 12, 13, 14, 15 and/or 16, or

b3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 10 and/or 215, preferably to SEQ ID NO: 10, or

b4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 10 and/or 215, preferably of SEQ ID NO: 10, or

b5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 10 and/or 215, preferably of

SEQ ID NO: 10 per primer, or

b6) a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a pamamycin producing organism, or

b7) a polynucleotide of at least two of b1 ), b2), b3), b4), b5) and b6);

c) an expression cassette comprising

c1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or

100% identical to SEQ ID NO: 18 and/or 223, preferably toSEQ ID NO: 18, or

c2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18, 19, 20, 21 , 22, 23, 223, 224, 225, 226, 227 and/or 228, preferably to

SEQ ID NO: 18, 19, 20, 21 , 22 and/or 23, or

c3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17 and/or 222, preferably to SEQ ID NO: 17, or

c4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 17 and/or 222, preferably of SEQ ID NO: 17, or

c5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 17 and/or 222, preferably of

SEQ ID NO: 17 per primer, or

c6) a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a pamamycin producing organism, or

c7) a polynucleotide of at least two of c1 ), c2), c3), c4), c5) and c6);

d) an expression cassette comprising

d1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25 and/or 230, preferably to SEQ ID NO: 25, or

d2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, 26, 27, 28, 29, 30, 230, 231 , 232, 233, 234 and/or 235, preferably to SEQ ID NO: 25, 26, 27, 28, 29 and/or 30, or

d3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 24 and/or 229, preferably to SEQ ID NO: 24, or

d4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 24 and/or 229, preferably of SEQ ID NO: 24, or

d5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 24 and/or 229, preferably of SEQ ID NO: 24 per primer, or

6) a polynucleotide of d1), d2), d3), d4) and/or d5) being obtainable from a pamamycin producing organism, or

7) a polynucleotide of at least two of d 1 ), d2), d3), d4), d5) and d6);

n expression cassette comprising

1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 237, 349, 356, 363, 370 and/or 377, preferably to SEQ ID NO: 32 and/or 237, more preferred to SEQ ID NO: 32, or

2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 33, 34, 35, 36, 37, 237, 238, 239, 240, 241 , 242, 249, 350, 351 , 352, 353, 354, 356, 357, 358, 359, 360, 361 , 363, 364, 365, 366, 367, 368, 369, 371 , 372, 373, 374, 375, 377, 378, 379, 380, 381 and/or 382, preferably to SEQ ID NO: 32, 33, 34, 35, 36, 37, 237, 238, 239, 240, 241 , 242, more preferred to SEQ ID NO: 32, 33, 34, 35, 36 and/or 37, or

3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, preferably to SEQ ID NO: 31 and/or 236, more preferred to SEQ ID NO: 31 , or

4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, preferably of SEQ ID NO: 31 and/or 236, more preferred of SEQ ID NO: 31 , or

5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, preferably of SEQ ID NO: 31 and/or 236, more preferred of SEQ ID NO: 31 per primer, or

6) a polynucleotide of e1), e2), e3), e4) and/or e5) being obtainable from a pamamycin producing organism, or

7) a polynucleotide of at least two of e1 ), e2), e3), e4), e5) and e6);

an expression cassette comprising

) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39 and/or 244, preferably to SEQ ID NO: 39, or

) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, 40, 41 , 42, 43, 44, 244, 245, 246, 247, 248 and/or 249, preferably to SEQ ID NO: 39, 40, 41 , 42, 43 and/or 44, or f3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38 and/or 243, preferably to SEQ ID NO: 38, or

f4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 38 and/or 243, preferably of

SEQ ID NO: 38, or

f5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 38 and/or 243, preferably of SEQ ID NO: 38 per primer, or

f6) a polynucleotide of f1), f2), f3), f4) and/or f5) being obtainable from a

pamamycin producing organism, or

f7) a polynucleotide of at least two of f1 ), f2), f3), f4), f5) and f6);

g) an expression cassette comprising

g1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46 and/or 251 , preferably to SEQ ID NO: 46, or

g2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:

46, 47, 48, 49, 50, 51 , 251 , 252, 253, 254, 255 and/or 256, preferably to

SEQ ID NO: 46, 47, 48, 49, 50 and/or 51 , or

g3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45 and/or 250, preferably to SEQ ID NO: 45, or

g4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 45 and/or 250, preferably of

SEQ ID NO: 45, or

g5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 45 and/or 250, preferably of SEQ ID NO: 45 per primer, or

g6) a polynucleotide of g1), g2), g3), g4) and/or g5) being obtainable from a pamamycin producing organism, or

g7) a polynucleotide of at least two of g 1 ), g2), g3), g4), g5) and g6);

h) an expression cassette comprising

hi ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, 258, 384, 391 , 398, 405 and/or 412, preferably to SEQ ID NO: 53, 258 and/or 384, more preferred to SEQ ID

NO: 53, or

h2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, 54, 55, 56, 57, 58, 384, 385, 386, 387, 388, 389, 391 , 392, 393, 394, 395, 396, 398, 399, 400, 401 , 402, 403, 405, 406, 407, 408, 409, 410, 412, 413, 414, 415, 416 and/or 417, preferably to SEQ ID NO: 53, 54, 55, 56, 57, 58, 384, 385, 386, 387, 388 and/or 389, more preferred to SEQ ID NO:

53, 54, 55, 56, 57 and/or 58, or

h3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , preferably to SEQ ID NO: 52, 257 and/or 383, more preferred to SEQ ID NO: 52, or

h4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , preferably of SEQ ID NO: 52, 257 and/or 383, more preferred of SEQ ID NO: 52, or

h5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , preferably of SEQ ID NO: 52, 257 and/or 383, more preferred of SEQ ID NO: 52 per primer, or

h6) a polynucleotide of hi), h2), h3), h4) and/or h5) being obtainable from a pamamycin producing organism, or

h7) a polynucleotide of at least two of h 1 ), h2), h3), h4), h5) and h6);

an expression cassette comprising

11 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or

100% identical to SEQ ID NO: 60 and/or 265, preferably to SEQ ID NO: 60, or

12) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, 61 , 62, 63, 64, 65, 265, 266, 267, 268, 269 and/or 270, preferably to

SEQ ID NO: 60, 61 , 62, 63, 64 and/or 65, or

13) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 59 and/or 264, preferably to SEQ ID NO: 59, or

i4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 59 and/or 264, preferably of SEQ ID NO: 59, or

15) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 59 and/or 264, preferably of

SEQ ID NO: 59 per primer, or

16) a polynucleotide of i 1 ) , i2), i3), i4) and/or i5) being obtainable from a pamamycin producing organism, or

i7) a polynucleotide of at least two of i 1 ) , i2), i3), i4), i5) and i6);

j) an expression cassette comprising

j1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or

100% identical to SEQ ID NO: 67 and/or 272, preferably to SEQ ID NO: 67, or

j2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, 68, 69, 70, 71 , 72, 272, 273, 274, 275, 276 and/or 277, preferably to

SEQ ID NO: 67, 68, 69, 70, 71 and/or 72, or

j3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66 and/or 271 , preferably to SEQ ID NO: 66, or

j4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 66 and/or 271 , preferably of SEQ ID NO: 66, or

j5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 66 and/or 271 , preferably of

SEQ ID NO: 66 per primer, or

j6) a polynucleotide of j 1 ) , j2), j3), j4) and/or j5) being obtainable from a

pamamycin producing organism, or

j7) a polynucleotide of at least two of j 1 ) , j2), j3), j4), j5) and j6);

k) an expression cassette comprising

k1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74 and/or 279, preferably to SEQ ID NO: 74, or

k2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, 75, 76, 77, 78, 79, 279, 280, 281 , 282, 283 and/or 284, preferably to SEQ ID NO: 74, 75, 76, 77, 78 and/or 79, or

k3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 73 and/or 278, preferably to SEQ ID NO: 73, or

k4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 73 and/or 278, preferably of SEQ ID NO: 73, or

k5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 73 and/or 278, preferably of SEQ ID NO: 73 per primer, or

I a polynucleotide of k1), k2), k3), k4) and/or k5) being obtainable from a pamamycin producing organism, or

I a polynucleotide of at least two of k1 ), k2), k3), k4), k5) and k6);

expression cassette comprising

a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 and/or 286, preferably to SEQ ID NO: 81 , or

a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , 82, 83, 84, 85, 86, 286, 287, 288, 289, 290 and/or 291 , preferably to SEQ ID NO: 81 , 82, 83, 84, 85 and/or 86, or

a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 80 and/or 285, preferably to SEQ ID NO: 80, or

a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 80 and/or 285, preferably of SEQ ID NO: 80, or

a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 80 and/or 285, preferably of SEQ ID NO: 80 per primer, or

a polynucleotide of 11), I2), I3), I4) and/or I5) being obtainable from a pamamycin producing organism, or

a polynucleotide of at least two of 11 ), I2), I3), I4), I5) and I6);

expression cassette comprising

) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88 and/or 293, preferably to SEQ ID NO: 88, or

) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, 89, 90, 91 , 92, 93, 293, 294, 295, 296, 297 and/or 298, preferably to SEQ ID NO: 88, 89, 90, 91 , 92 and/or 93, or

) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87 and/or 292, preferably to SEQ ID NO: 87, or

) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 87 and/or 292, preferably of SEQ ID NO: 87, or

) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 87 and/or 292, preferably of SEQ ID NO: 87 per primer, or

m6) a polynucleotide of ml ), m2), m3), m4) and/or m5) being obtainable from a pamamycin producing organism, or

m7) a polynucleotide of at least two of ml ), m2), m3), m4), m5) and m6);

n) an expression cassette comprising

n1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 95 and/or 300, preferably to SEQ ID NO:

95, or

n2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 95, 96, 97, 98, 99, 100, 300, 301 , 302, 303, 304 and/or 305, preferably to SEQ ID NO: 95, 96, 97, 98, 99 and/or 100, or

n3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 94 and/or 299, preferably to SEQ ID NO: 94, or

n4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 94 and/or 299, preferably of

SEQ ID NO: 94, or

n5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 94 and/or 299, preferably of SEQ ID NO: 94 per primer, or

n6) a polynucleotide of n1), n2), n3), n4) and/or n5) being obtainable from a pamamycin producing organism, or

n7) a polynucleotide of at least two of n 1 ), n2), n3), n4), n5) and n6);

o) an expression cassette comprising

o1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102 and/or 307, preferably to SEQ ID NO: 102, or

02) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:

102, 103, 104, 105, 106, 107, 307, 308, 309, 310, 31 1 and/or 312, preferably to SEQ ID NO: 102, 103, 104, 105, 106 and/or 107, or

03) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 101 and/or 306, preferably to SEQ ID NO: 101 , or

04) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 101 and/or 306, preferably of SEQ ID NO: 101 , or

05) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 101 and/or 306, preferably of SEQ ID NO: 101 per primer, or

06) a polynucleotide of o1), o2), o3), o4) and/or o5) being obtainable from a pamamycin producing organism, or

07) a polynucleotide of at least two of o1 ), o2), o3), o4), o5) and 06); p) an expression cassette comprising,

p1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109 and/or 314, preferably to SEQ ID NO: 109, or

p2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:

109, 1 10, 1 1 1 , 1 12, 1 13, 1 14, 314, 315, 316, 317, 318 and/or 319, preferably to SEQ ID NO: 109, 1 10, 1 1 1 , 1 12, 1 13 and/or 1 14, or p3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 108 and/or 313, preferably to SEQ ID NO: 108, or

p4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 108 and/or 313, preferably of

SEQ ID NO: 108, or

p5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 108 and/or 313, preferably of SEQ ID NO: 108 per primer, or

p6) a polynucleotide of p1), p2), p3), p4) and/or p5) being obtainable from a pamamycin producing organism, or

p7) a polynucleotide of at least two of p1 ), p2), p3), p4), p5) and p6); wherein at least one of the expression cassettes of a) to p) is a recombinant expression cassette, or

wherein at least one of the expression cassettes is comprised by a recombinant polynucleotide.

In further embodiments, the microorganisms comprise at least one expression cassette as described in a1) to t7) below.

a) an expression cassette comprising

a1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, or

a2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, 5, 6, 7, 8, and/or 9, or

a3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3, or a4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 3, or

a5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 3 per primer, or

a6) a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a

pamamycin producing organism, or

a7) a polynucleotide of at least two of a1 ), a2), a3), a4), a5) and a6); an expression cassette comprising

b1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 , or

b2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 , 12, 13,

14, 15 and/or 16, or

b3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 10, or b4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 10, or

b5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 10 per primer, or

b6) a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a

pamamycin producing organism, or

b7) a polynucleotide of at least two of b1 ), b2), b3), b4), b5) and b6); an expression cassette comprising

c1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18, or

c2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18, 19, 20,

21 , 22 and/or 23, or

c3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17, or c4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 17, or c5 a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 17 per primer, or

c6 a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a

pamamycin producing organism, or

c7 a polynucleotide of at least two of c1 ), c2), c3), c4), c5) and c6);

an expression cassette comprising

d1 a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, or

d2 a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, 26, 27, 28, 29 and/or 30, or

d3 a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 24, or d4 a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 24, or

d5 a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 24 per primer, or

d6 a polynucleotide of d1), d2), d3), d4) and/or d5) being obtainable from a

pamamycin producing organism, or

d7 a polynucleotide of at least two of d1 ), d2), d3), d4), d5) and d6); an expression cassette comprising

e1 a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, or

e2 a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 33, 34, 35, 36 and/or 37, or

e3 a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31 , or e4 a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 31 , or

e5 a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 31 per primer, or

e6 a polynucleotide of e1), e2), e3), e4) and/or e5) being obtainable from a

pamamycin producing organism, or

e7 a polynucleotide of at least two of e1 ), e2), e3), e4), e5) and e6); an expression cassette comprising

f1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, or

f2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, 40, 41 ,

42, 43 and/or 44, or

f3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38, or f4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 38, or

f5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 38 per primer, or

f6) a polynucleotide of f1), f2), f3), f4) and/or f5) being obtainable from a

pamamycin producing organism, or

f7) a polynucleotide of at least two of f1 ), f2), f3), f4), f5) and f6); an expression cassette comprising

g1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46, or

g2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46, 47, 48,

49, 50 and/or 51 , or

g3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45, or g4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 45, or

g5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 45 per primer, or

g6) a polynucleotide of g1), g2), g3), g4) and/or g5) being obtainable from a

pamamycin producing organism, or

g7) a polynucleotide of at least two of g 1 ), g2), g3), g4), g5) and g6); an expression cassette comprising

hi ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, or

h2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, 54, 55, 56, 57 and/or 58, or

a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 52, or a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 52, or

a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 52 per primer, or

a polynucleotide of hi), h2), h3), h4) and/or h5) being obtainable from a pamamycin producing organism, or

a polynucleotide of at least two of h 1 ), h2), h3), h4), h5) and h6); expression cassette comprising

a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, or

a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, 61 , 62, 63, 64 and/or 65, or

a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 59, or a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 59, or

a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 59 per primer, or

a polynucleotide of i 1 ) , i2), i3), i4) and/or i5) being obtainable from a pamamycin producing organism, or

a polynucleotide of at least two of i 1 ), i2), i3), i4), i5) and i6); expression cassette comprising

a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, or

a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, 68, 69, 70, 71 and/or 72, or

a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66, or a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 66, or

a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 66 per primer, or

j6) a polynucleotide of j 1 ) , j2), j3), j4) and/or j5) being obtainable from a pamamycin producing organism, or

j7) a polynucleotide of at least two of j 1 ) , j2), j3), j4), j5) and j6); an expression cassette comprising

k1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, or

k2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, 75, 76,

77, 78 and/or 79, or

k3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 73, or k4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 73, or

k5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 73 per primer, or

k6) a polynucleotide of k1), k2), k3), k4) and/or k5) being obtainable from a

pamamycin producing organism, or

k7) a polynucleotide of at least two of k1 ), k2), k3), k4), k5) and k6); an expression cassette comprising

11 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , or

12) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , 82, 83, 84, 85 and/or 86, or

13) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 80, or

14) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 80, or

15) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 80 per primer, or

16) a polynucleotide of 11), I2), I3), I4) and/or I5) being obtainable from a pamamycin producing organism, or

17) a polynucleotide of at least two of 11 ), I2), I3), I4), I5) and I6); an expression cassette comprising

ml a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, or

m2 a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, 89, 90, 91 , 92 and/or 93, or

m3 a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87, or m4 a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 87, or

m5 a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 87 per primer, or

m6 a polynucleotide of ml), m2), m3), m4) and/or m5) being obtainable from a pamamycin producing organism, or

m7 a polynucleotide of at least two of ml ), m2), m3), m4), m5) and m6); expression cassette comprising

a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 95, or

a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 95, 96, 97, 98, 99 and/or 100, or

a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 94, or a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 94, or

a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 94 per primer, or

a polynucleotide of n1), n2), n3), n4) and/or n5) being obtainable from a pamamycin producing organism, or

a polynucleotide of at least two of n 1 ), n2), n3), n4), n5) and n6); an expression cassette comprising

01 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102, or

02) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102, 103, 104, 105, 106 and/or 107, or

03) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 101 , or

04) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 101 , or

05) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 101 per primer, or

06) a polynucleotide of o1), o2), o3), o4) and/or o5) being obtainable from a

pamamycin producing organism, or

07) a polynucleotide of at least two of o1 ), o2), o3), o4), o5) and 06); an expression cassette comprising,

p1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, or

p2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, 1 10,

1 1 1 , 1 12, 1 13 and/or 1 14, or

p3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 108, or

p4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 108, or

p5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 108 per primer, or

p6) a polynucleotide of p1), p2), p3), p4) and/or p5) being obtainable from a

pamamycin producing organism, or

p7) a polynucleotide of at least two of p1 ), p2), p3), p4), p5) and p6); at least one expression cassette comprising

q1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, or

q2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17,

1 18, 1 19, 120 and/or 121 , or

q3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15, or q4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 1 15, or

q5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 per primer, or

q6) a polynucleotide of q1), q2), q3), q4) and/or q5) being obtainable from a

pamamycin producing organism, or

q7) a polynucleotide of at least two of q 1 ), q2), q3), q4), q5) and q6); at least one expression cassette comprising

r1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, or

r2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, 131 ,

132, 133, 134 and/or 135, or

r3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 129, or

r4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 129, or

r5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 129 per primer, or

r6) a polynucleotide of r1), r2), r3), r4) and/or r5) being obtainable from a

pamamycin producing organism, or

r7) a polynucleotide of at least two of r1 ), r2), r3), r4), r5) and r6); at least one expression cassette comprising

s1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, or

s2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, 138,

139, 140, 141 and 142, or

s3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 136, or

s4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 136, or

s5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 136 per primer, or

s6) a polynucleotide of s1), s2), s3), s4) and/or s5) being obtainable from a

pamamycin producing organism, or

s7) a polynucleotide of at least two of s1 ), s2), s3), s4), s5) and s6); t) at least one expression cassette comprising

t1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123, or

t2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123, 124,

125, 126, 127 and/or 128, or

t3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 122, or

t4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 122, or

t5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 122 per primer, or

t6) a polynucleotide of t1), t2), t3), t4) and/or t5) being obtainable from a

pamamycin producing organism, or

t7) a polynucleotide of at least two of t1 ), t2), t3), t4), t5) and t6);

Preferred embodiments define the polypeptides of a) to p), or the polypeptides of a) to s) as applicable, according to the criteria of a1) to p7) or a1) to s7), respectively, as described above, including preferred combinations and sequence identity regions of these criteria described above.

The recombinant microorganisms may comprise single expression cassettes of a1) to t7), or may comprise combinations of these expression cassettes, for example at least one expression cassette selected from a1) to a7), at least one expression cassette selected from b1) to b7), at least one expression cassette selected from c1) to c7), at least one expression cassette selected from d1 ) to d7), at least one expression cassette selected from e1) to e7), at least one expression cassette selected from f1) to f7), at least one expression cassette selected from g1 ) to g7), at least one expression cassette selected from hi) to h7), at least one expression cassette selected from i 1 ) to i7), at least one expression cassette selected from j1 ) to j7), at least one expression cassette selected from k1) to k7), at least one expression cassette selected from 11 ) to I7), at least one expression cassette selected from ml) to m7), at least one expression cassette selected from n1 ) to n7), at least one expression cassette selected from o1 ) to o7) and at least one expression cassette selected from p1 ) to p7). Other combinations are also possible, e.g. combinations which comprise at least one expression cassette for every polypeptide encoding region comprised by SEQ ID NO: 1 or 2, or for every polypeptide encoding region comprised by SEQ ID NO: 2, and at least one expression cassette selected from q1) to q7), at least one expression cassette selected from r1 ) to r7), at least one expression cassette selected from s1) to s7) and at least one expression cassette selected from t1 ) to t7).

An individual expression cassette may encode only one polypeptide having one of the above mentioned activities, but may also have an operon like structure, so that it encodes more than one polypeptide having the same or different activities selected from the ones described above.

The expression cassettes may be recombinant, e.g. by comprising heterologous promoter or terminator sequences or artificial variants of the polypeptides having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS pamR1 , pamW, pamR2 or pamH activity, or articial variants of the

polynucleotides encoding these polypeptides. However, the expression cassette or expression cassette may also represent fragments of SEQ ID NO: 1 and/or 2, which are comprised in recombinant polynucleotides, for example recombinant polynucleotides which combine several expression cassettes comprised by SEQ ID NO: 1 and/or 2 in a different order, number and/or orientation. A person skilled in the art will know that there are many different designs of those recombinant polynucleotides possible. The simplest design will be to create fragments of SEQ ID NO: 1 and/or 2 or to combine these fragments to create new polynucleotides. Preferred emdodiments of these polynucleotides are the polynucleotides depicted in Figure 1.

The different variants on how to design the respective expression cassettes as described here will also apply to the expression cassettes described for any of the recombinant microorganisms described above. A person skilled in the art will understand, that these varaints are not limiting, and will be able to create expression cassettes of different design. These recombinant microorganisms comprising expression cassettes of a1) to p7) or comprise expression cassettes a1 ) to t7) may have the capacity to produce at least one pamamycin, preferably at least pamamycin 607 or pamamycin 621 or both, or may not have the capacity to produce pamamycin. Guidance on which polypeptide activities need to be present in a pamamycin producing microorganism can be found in the disclosure of Figures 1 and 2.

Accordingly, the invention comprise recombinant microorganism comprising

i) at least one polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2, preferably 2,

ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2, preferably 2, and comprising at least one expression cassette for each one of a) to p)

a) at least one of a1), a2), a3), a4), a5), a6) or a7) as described above, and b) at least one of b1), b2), b3), b4), b5), b6) or b7) as described above, and c) at least one of c1 ), c2), c3), c4), c5), c6) or c7) as described above, and d) at least one of d1), d2), d3), d4), d5), d6) or d7) as described above, and e) at least one of e1 ), e2), e3), e4), e5), e6) or e7) as described above, and f) at least one of f 1 ), f2), f3), f4), f5), f6) or f7) as described above, and

g) at least one of g 1 ), g2), g3), g4), g5), g6) or g7) as described above, and h) at least one of hi), h2), h3), h4), h5), h6) or h7) as described above, and i) at least one of i 1 ) , i2), i3), i4), i5), i6) or i7) as described above, and

j) at least one of j 1 ) , j2), j3), j4), j5), j6) or j7) as described above, and

k) at least one of k1 ), k2), k3), k4), k5), k6) or k7) as described above, and

I) at least one of 11 ), I2), I3), I4), I5), I6) or I7) as described above, and

m) at least one of ml ), m2), m3), m4), m5), m6) or m7) as described above, and n) at least one of n1), n2), n3), n4), n5), n6) or n7) as described above, and

0) at least one of o1), o2), o3), o4), o5), 06) or o7) as described above, and p) at least one of p1), p2), p3), p4), p5), p6) or p7) as described above;

iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments

comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of

1) or ii), if the fragments are combined.

The recombinant microorganisms may also comprise at least one expression cassette comprising

q) at least one of q1), q2), q3), q4), q5), q6) or q7) as described above, or r) at least one of r1 ), r2), r3), r4), r5), r6) or r7) as described above, or

s) at least one of s1 ), s2), s3), s4), s5), s6) or s7) as described above, or t) at least one of t1), t2), t3), t4), t5), t6) or t7) as described above.

Preferably the recombinant microorganism comprise at least one expression cassette comprising

q) at least one of q 1 ), q2), q3), q4), q5), q6) or q7) as described above.

This embodiment is particularly preferred, in case the recombinant microorganism comprises expression cassettes for polypeptides having having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS pamR1 , pamW, pamR2 or pamH activity, which comprise promoter sequences comprised in SEQ ID NO: 1 and/or 2.

Also preferred are recombinant microorganisms comprising at least one expression cassette comprising

s) at least one of s1 ), s2), s3), s4), s5), s6) or s7) as described above.

The recombinant microorganism described herein, including man-made mutants of organisms having the capacity to produce pamamycin. These man-made mutants have preferably a higher capacity to produce at least one of the pamamycins described above, preferably at least one of pamamycin 607 and/or 612. These man-made mutants are preferably used to isolate new sequence variants of the polypeptides and polynucleotides described above and/or are used in methods to produce pamamycin. The recombinant microorganisms, including man-made mutants of organisms which naturally have the capacity ot produce at least one pamamycin, as described above may or may not be able to produce at least one pamamycin.

The recombinant microorganisms do preferably belong to a species as described above, more preferred they belong to the group of Streptomyces, e.g. the Streptomyces species described above, in particular to the pamamicin producing species described above.

Preferably the recombinant microorganisms are able to produce at least one pamamycin, in particular if they are man-made mutants of pamamycin producing organisms. Pamamycin producing recombinant microorganisms are preferably used in methods which involve the production of pamamycin by such microorganisms.

Accordingly, the invention comprises methods for the production of pamamycin, preferably for the production of pamamycin 607 or pamamycin 607, or both, comprising the steps of i) cultivating a recombinant microorganism having the capacity to produce pamamycin under conditions which allow for the production of pamamycin by said recombinant microorganism;

ii) obtaining produced pamamycin.

The recombinant microorganism preferably comprises:

a) at least one polypeptide having pamC activity, and

b) at least one polypeptide having pamG activity, and

c) at least one polypeptide having pamF activity, and

d) at least one polypeptide having pamA activity, and

e) at least one polypeptide having pamB activity, and

f) at least one polypeptide having pamD activity, and

g) at least one polypeptide having pamE activity, and

h) at least one polypeptide having pamO activity, and

i) at least one polypeptide having pamK activity, and

j) at least one polypeptide having pamJ activity, and

k) at least one polypeptide having pamM activity, and

I) at least one polypeptide having pamN activity, and

m) at least one polypeptide having pamL activity, and

n) at least one polypeptide having pamX activity, and

o) at least one polypeptide having pamY activity, and

P) at least one polypeptide having pamS activity,

wherein at least one of those activities a) to p) is up-regulated, or

comprising the activities of a) to p) and comprising

q) an up-regulated polypeptide having pamR1 activity, or

r) a down-regulated polypeptide having pamR2 activity, or

s) an up-regulated polypeptide having pamW activity, or t) a down-regulated polypeptide having pamH activity, or

u) a combination of at least two of q), r) s) and t), or

v) at least one up-regulated activities of a) to p) and at least one of q) to u). The polypeptides of a) to t) are preferably provided by at least one expression cassette for each one of a) to t)

a) at least one of a1), a2), a3), a4), a5), a6) or a7) as described above, and

b) at least one of b1 ), b2), b3), b4), b5), b6) or b7) as described above, and

c) at least one of c1 ), c2), c3), c4), c5), c6) or c7) as described above, and

d) at least one of d1), d2), d3), d4), d5), d6) or d7) as described above, and

e) at least one of e1 ), e2), e3), e4), e5), e6) or e7) as described above, and

f) at least one of f1), f2), f3), f4), f5), f6) or f7) as described above, and

g) at least one of g1), g2), g3), g4), g5), g6) or g7) as described above, and

h) at least one of hi), h2), h3), h4), h5), h6) or h7) as described above, and

i) at least one of i 1 ) , i2), i3), i4), i5), i6) or i7) as described above, and

j) at least one of j 1 ) , j2), j3), j4), j5), j6) or j7) as described above, and

k) at least one of k1 ), k2), k3), k4), k5), k6) or k7) as described above, and

I) at least one of 11 ), I2), I3), I4), I5), I6) or I7) as described above, and

m) at least one of ml ), m2), m3), m4), m5), m6) or m7) as described above, and n) at least one of n1), n2), n3), n4), n5), n6) or n7) as described above, and

o) at least one of o1 ), o2), o3), o4), o5), 06) or o7) as described above, and

P) at least one of p1), p2), p3), p4), p5), p6) or p7) as described above;

q) at least one of q1), q2), q3), q4), q5), q6) or q7) as described above, or

r) at least one of r1 ), r2), r3), r4), r5), r6) or r7) as described above, or

s) at least one of s1 ), s2), s3), s4), s5), s6) or s7) as described above, or

t) at least one of t1 ), t2), t3), t4), t5), t6) or t7) as described above.

The expression cassettes may be located on

i) at least one polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID

NO: 1 and/or 2, preferably on at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2,

ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2, preferably on at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2, and comprising at least one expression cassette for each one of a) to p) iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments

comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of i) or ii), if the fragments are combined.

Alternative means to produce pamamamycin, preferably pamamycin 607 and/or 621 is to produce first the compounds S-chain and L-chain and to provide microorganisms comprising polypeptides for the activities pamK, pamJ and pamL with the produced compounds S-chain and L-chain.

Accordingly, the invention comprises a method for the production of pamamycin comprising the steps of

a) cultivating a recombinant microorganism comprising

at least one polypeptide having pamK activity, and

at least one polypeptide having pamJ activity, and

at least one polypeptide having pamL activity,

b) providing the recombinant microorganism with compounds S-chain and L-chain, c) cultivating a recombinant microorganism under conditions which allow for the production of pamamycin by said recombinant microorganism and;

d) obtaining produced pamamycin, preferably obtaining pamamycin 607 and/or pamamycin 621 .

Preferably, the polypeptide having pamK activity is provided by

i 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D

NO: 60 and/or 265, or

\2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ I D NO: 60, 61 , 62, 63, 64, 65,

265, 266, 267, 268, 269 and/or 270, or

13) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 59 and/or 264, or

14) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ I D NO: 59 and/or 264, or

15) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ I D NO: 59 and/or 264 per primer, or

16) a polypeptide of i 1 ) and/or i2) and comprising the amino acids acids F33, C159, G314 and at least one of G346 and G348 using the alignment in Figure 1 1 as reference, or

17) a polypeptide of at least two of i 1 ) , i2), i3), i4), i5) and i6);

More preferred, the polypeptide having pamK activity is provided by i 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 60 and/or 265, or

\2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 60, 61 , 62, 63, 64, 65,

265, 266, 267, 268, 269 and/or 270, or

i3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 59 and/or 264, or

i4) a polypeptide of at least two of i 1 ), i2) and i3).

Preferably, the polypeptide having pamJ activity is provided by

j 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 67 and/or 272, or

j2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 67, 68, 69, 70, 71 , 72, 272, 273, 274, 275, 276 and/or 277, or

j3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 66 and/or 271 , or

j4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ I D NO: 66 and/or 271 , or

j5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ I D NO: 66 and/or 271 per primer, or

j6) a polypeptide of j 1 ) and/or j2) and comprising the amino acids G 14, G246, G252, G256 and H257 using the alignment in Figure 12 as reference, or j7) a polypeptide of at least two of j 1 ) , j2), j3), j4), j5) and j6).

More preferred, the polypeptide having pamJ activity is provided by

j 1 ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D

NO: 67 and/or 272, or

j2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 67, 68, 69, 70, 71 , 72, 272, 273, 274, 275, 276 and/or 277, or

j3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 66 and/or 271 , or j4) a polypeptide of at least two of j 1 ) , j2) and j3).

Preferably, the polypeptide having pamL activity is provided by

ml ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 88 and/or 293, or

m2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, 89, 90, 91 , 92, 93, 293, 294, 295, 296, 297 and/or 298, or

m3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%,

84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87 and/or 292, or

m4) a polypeptide expressed from a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 87 and/or 292, or

m5) a polypeptide expressed from a polynucleotide which is obtainable via

PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 87 and/or 292 per primer, or

m6) a polypeptide of ml ) and/or m2) and comprising the amino acids L82, V93,

T214, S215, G216, G219, K222, E352, G370, G394, Y410, T438, G439, D430, R445, G454, E463, P520, P530, G535, K536 and at least one of T217 and T218 using the alignment in Figure 15 as reference, or m7) a polypeptide of at least two of ml ), m2), m3), m4), m5) and m6).

More preferred, the polypeptide having pamL activity is provided by

ml ) a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88 and/or 293, or

m2) a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%,

97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, 89, 90, 91 , 92, 93, 293, 294, 295, 296, 297 and/or 298, or

m3) a polypeptide expressed from a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87 and/or 292, or

m4) a polypeptide of at least two of ml ), m2) and m3).

The recombinant microorganisms suitable to be used in those methods are also provided by the invention.

Accordingly, further embodiments comprise recombinant microorganism comprising

a) at least one polypeptide having pamK activity, and

b) at least one polypeptide having pamJ activity, and c) at least one polypeptide having pamL activity,

wherein at least one of of a), b) or c) is a recombianant polypeptide, or is expressed from a recombinant expression cassette or is expressed from an expression cassette located on a recombinant polynucleotide.

In particular preferred are recombinant microorganisms which comprise polypeptides having pamK, pamJ or pamL activity as described above for the description of the respective method. The produced pamamycin can be hydrolysed in order to produce compounds of the

Formula IV or Formula V or both,

Accordingly, the invention comprises a method for the production of compounds of the Formula IV or Formula V comprising the steps of:

i) cultivating a recombinant microorganism as having a capacity to produce pamamycin under conditions which allow for the production of pamamycin by said recombinant microorganism;

ii) obtaining produced pamamycin, preferably obtaining pamamycin 607 and/or

pamamycin 621 ,

iii) hydrolysing pamamycin, preferably hydrolysing pamamycin 607 and/or pamamycin 621 and

iv) obtaining compounds of Formula IV or obtaining compounds of Formula V or

obtaining compounds of Formula IV and Formula V.

Preferred embodiments define the polypeptides of a) to p), or the polypeptides of a) to t) as applicable, according to the criteria of a1) to p7) or a1) to t7), respectively, as described above, including preferred combinations and sequence identity regions of these criteria described in the different embodiments of the recombinant microorganisms disclosed above. The produced pamamycin and/or compounds of Formula IV or Formula V or both may be obtained from the culture broth or from the recombinant organisms or both, preferably they are obtained from the culture broth.

The produced pamamycin and/or compounds of Formula IV or Formula V or both may be obtained in the form of their agriculturally and/or pharmacologically acceptable salts.

Preferably they are obtained in the form of their agriculturally acceptable salts.

The invention also comprises polynucleotides, expression cassettes and vectors comprising any one of the polynucleotides described above, and host cells comprising such vectors or polynucleotides.

Suitable conditions which allow for production of pamamycin depend on the recombinant microorganism as well as the promoters used to control the production of polypeptides having having pamC, pamG, pamF, pamA, pamB, pamD, pamE, pamO, pamK, pamJ, pamM, pamN, pamL, pamX, pamY, pamS pamR1 , pamW, pamR2 or pamH activity. These conditions and how to select them are well known to those skilled in the art.

The term " cultivating" as used herein refers maintaining and growing the recombinant microorganism under culture conditions which allow the cells to produce pamamycin. This implies that the polynucleotides of the present invention are expressed in the recombinant microorganism so that the encoded polypeptides are present in the recombinant

microorganism in a biologically active form. Suitable culture conditions for cultivating the recombinant microorganism are described in more detail in the accompanying Examples below.

In particular, recombinant microorganisms of the present invention can be cultured using, for example, glucose, sucrose, maltose, honey, dextrin, starch, glycerol, molasses, animal or vegetable oils and the like as the carbon source for the culture medium. Furthermore, soybean flour, wheat germ, corn steep liquor, cotton seed waste, meat extract,

polypeptone, malt extract, yeast extract, ammonium sulfate, sodium nitrate, urea and the like can be used for the nitrogen source. The addition of inorganic salts which can produce sodium, potassium, calcium, magnesium, cobalt, chlorine, phosphoric acid (di-potassium hydrogen phosphate and the like), sulfuric acid (magnesium sulfate and the like) and other ions as required is also effective. Furthermore, various vitamins such as thiamine (thiamine hydrochloride and the like), amino acids such as glutamine (sodium glutamate and the like), asparagine (DL-asparagine and the like), trace nutrients such as nucleotides and the like, and selection drugs such as antibiotics and the like can also be added as required.

Moreover, organic substances and inorganic substances can be added appropriately to assist the growth of the microorganism and promote the production of pamamycin and/or to promote the preduction of pamamycin precursors. The pH of the culture medium is, for example, of the order of pH 4.5 to pH 8. The culturing can be carried out with a method such as the solid culturing method under aerobic conditions, the concussion culturing method, the air-passing agitation culturing method or the deep aerobic culturing method, but the deep aerobic culturing method is the most suitable. The appropriate temperature for culturing is from 15°C to 40°C, but in many cases growth occurs in the range from 20°C to 30°C. Usually a temperature of about 28°C is used. The production of pamamycin or precursors of pamamycin differs according to the culture medium and culturing conditions, or the host which is being used, but with any culturing method the accumulation of pamamycin reaches a maximum generally in from 5 to 20 days. The culturing is stopped when the amount of pamamycin or precursors of pamamycin in the culture reaches its highest level and the target material is isolated from the culture and refined for isolating pamamycin or precursor of pamamycin from the culture material.

Examples for such conditions which allow for the production of pamamycin are disclosed in US4283391 , DE4134168 and DE4316836 which are included herein by reference in their entirety.

The term " obtaining" as used herein encompasses the provision of the cell culture including the recombinant microorganisms and the culture medium as well as the provision of purified or partially purified preparations thereof comprising pamamycin or a precursor thereof, preferably, in free form. More details on purification techniques can be found elsewhere herein below. The usual methods of extraction and refinement which are generally used in these circumstances, such as methods of isolation such as solvent extraction, methods involving ion exchange resins, adsorption or partition chromatography, gel filtration, dialysis, precipitation, crystallization and the like can be used either individually or in appropriate combinations. In particular, pamamycin can be isolated from a pamamycin containing medium or cell lysate, or from both, using any applicable method, for example but not excluding others, methods which have been described in the prior art for the isolation of pamamycin.

Further embodiments of the invention are recombinant polynucleotides, recombinant expression cassettes and vectors comprising any one of the polynucleotides described above, and host cells comprising such vectors or polynucleotides.

Recombinant polynucleotides and recombinant expression cassettes are polynucleotides and expression cassettes, which are modified by genetic engineering or comprise sequences derived from man-made mutations of naturally occurring polynucleotides. Thus, they have less than 100% sequence identity to polynucleotides of SEQ ID NOs: 1 , 2, 3, 10, 17, 24, 31 , 38, 45, 52, 59, 66, 73, 80, 87, 94, 101 or 108 and their naturally occurring variants. Recombinant expression cassettes have less than 100% sequence identity to fragments of SEQ ID NO: 1 or 2 and their naturally occurring variants comprising polynucleotides of SEQ ID NOs: 1 , 2, 3, 10, 17, 24, 31 , 38, 45, 52, 59, 66, 73, 80, 87, 94, 101 or 108 and their naturally occurring variants flanked by the respective promoter and terminator sequences directly flanking those polynucleotides in nature.

Hence, the invention comprises polynucleotides, expression cassettes, vectors and host cells comprising a polynucletotide encoding at least one of a) to v):

a) at least one polypeptide having pamC activity, and

b) at least one polypeptide having pamG activity, and

c) at least one polypeptide having pamF activity, and

d) at least one polypeptide having pamA activity, and

e) at least one polypeptide having pamB activity, and

f) at least one polypeptide having pamD activity, and

g) at least one polypeptide having pamE activity, and

h) at least one polypeptide having pamO activity, and

i) at least one polypeptide having pamK activity, and

j) at least one polypeptide having pamJ activity, and

k) at least one polypeptide having pamM activity, and

I) at least one polypeptide having pamN activity, and

m) at least one polypeptide having pamL activity, and n) at least one polypeptide having pamX activity, and

o) at least one polypeptide having pamY activity, and

p) at least one polypeptide having pamS activity,

wherein at least one of those activities a) to p) is up-regulated, or

comprising the activities of a) to p) and comprising

q) an up-regulated polypeptide having pamR1 activity, or

r) a down-regulated polypeptide having pamR2 activity, or

s) an up-regulated polypeptide having pamW activity, or

t) a down-regulated polypeptide having pamH activity, or

u) a combination of at least two of q), r) s) and t), or

v) at least one up-regulated activities of a) to p) and at least one of q) to u).

The polynucleotides, expression cassettes, vectors and host cells preferably comprise a polynucleotide of at least one of a) to t), being

a) at least one of a1 ), a2), a3), a4), a5), a6) or a7) as defined in

a1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4 and/or 209, preferably to SEQ ID NO: 4, or

a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, 5, 6, 7, 8, 9, 209, 210, 21 1 , 212, 213 and/or 214, preferably to SEQ ID NO: 4, 5, 6, 7, 8, and/or 9, or

a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3 and/or 208, preferably to SEQ ID NO: 3, or

a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 3 and/or 208, preferably of SEQ ID NO: 3, or

a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 3 and/or 208, preferably of SEQ ID NO: 3 per primer, or

a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from pamamycin producing organism, or

a7) a polynucleotide of at least two of a1 ), a2), a3), a4), a5) and a6); at least one of b1 ), b2), b3), b4), b5), b6) or b7) as defined

a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 and/or 216, preferably to SEQ ID NO: 1 1 , or b2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 , 12, 13, 14, 15, 16, 216, 217, 218, 219, 220 and/or 221 , preferably to SEQ ID NO: 1 1 , 12, 13, 14, 15 and/or 16, or

b3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 10 and/or 215, preferably to SEQ ID NO: 10, or

b4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 10 and/or 215, preferably of SEQ ID NO: 10, or

b5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 10 and/or 215, preferably of SEQ ID NO: 10 per primer, or

b6) a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a pamamycin producing organism, or

b7) a polynucleotide of at least two of b1 ), b2), b3), b4), b5) and b6); c) at least one of c1 ), c2), c3), c4), c5), c6) or c7) as defined in

c1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18 and/or 223, preferably toSEQ ID NO: 18, or

c2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:

18, 19, 20, 21 , 22, 23, 223, 224, 225, 226, 227 and/or 228, preferably to

SEQ ID NO: 18, 19, 20, 21 , 22 and/or 23, or

c3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17 and/or 222, preferably to SEQ ID NO: 17, or

c4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 17 and/or 222, preferably of

SEQ ID NO: 17, or

c5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 17 and/or 222, preferably of SEQ ID NO: 17 per primer, or

c6) a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a pamamycin producing organism, or

c7) a polynucleotide of at least two of c1 ), c2), c3), c4), c5) and c6);

d) at least one of d 1 ), d2), d3), d4), d5), d6) or d7) as defined in

d1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25 and/or 230, preferably to SEQ ID NO: 25, or

d2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:

25, 26, 27, 28, 29, 30, 230, 231 , 232, 233, 234 and/or 235, preferably to

SEQ ID NO: 25, 26, 27, 28, 29 and/or 30, or

d3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 24 and/or 229, preferably to SEQ ID NO: 24, or

d4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 24 and/or 229, preferably of

SEQ ID NO: 24, or

d5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 24 and/or 229, preferably of SEQ ID NO: 24 per primer, or

d6) a polynucleotide of d1), d2), d3), d4) and/or d5) being obtainable from a pamamycin producing organism, or

d7) a polynucleotide of at least two of d 1 ), d2), d3), d4), d5) and d6);

at least one of e1 ), e2), e3), e4), e5), e6) or e7) as defined in

e1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 237, 349, 356, 363, 370 and/or 377, preferably to SEQ ID NO: 32 and/or 237, more preferred to SEQ ID NO: 32, or

e2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 33, 34, 35, 36, 37, 237, 238, 239, 240, 241 , 242, 249, 350, 351 , 352, 353, 354, 356, 357, 358, 359, 360, 361 , 363, 364, 365, 366, 367, 368, 369, 371 , 372, 373, 374, 375, 377, 378, 379, 380, 381 and/or 382, preferably to SEQ ID NO: 32, 33, 34, 35, 36, 37, 237, 238, 239, 240, 241 , 242, more preferred to SEQ ID NO: 32, 33, 34, 35, 36 and/or 37, or

e3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, preferably to SEQ ID NO: 31 and/or 236, more preferred to SEQ ID NO: 31 , or

e4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, preferably of SEQ ID NO: 31 and/or 236, more preferred of SEQ ID NO: 31 , or

e5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 31 , 236, 348, 355, 362, 369 and/or 376, preferably of SEQ ID NO: 31 and/or 236, more preferred of SEQ ID NO: 31 per primer, or

a polynucleotide of e1), e2), e3), e4) and/or e5) being obtainable from a pamamycin producing organism, or

a polynucleotide of at least two of e1 ), e2), e3), e4), e5) and e6); f) at least one of f1), f2), f3), f4), f5), f6) or f7) as defined in

f1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39 and/or 244, preferably to SEQ ID NO: 39, or

f2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:

39, 40, 41 , 42, 43, 44, 244, 245, 246, 247, 248 and/or 249, preferably to

SEQ ID NO: 39, 40, 41 , 42, 43 and/or 44, or

f3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38 and/or 243, preferably to SEQ ID NO: 38, or

f4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 38 and/or 243, preferably of

SEQ ID NO: 38, or

f5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 38 and/or 243, preferably of SEQ ID NO: 38 per primer, or

f6) a polynucleotide of f1), f2), f3), f4) and/or f5) being obtainable from a

pamamycin producing organism, or

f7) a polynucleotide of at least two of f1 ), f2), f3), f4), f5) and f6);

g) at least one of g 1 ), g2), g3), g4), g5), g6) or g7) as defined in

g1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46 and/or 251 , preferably to SEQ ID NO: 46, or

g2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46, 47, 48, 49, 50, 51 , 251 , 252, 253, 254, 255 and/or 256, preferably to SEQ ID NO: 46, 47, 48, 49, 50 and/or 51 , or

g3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45 and/or 250, preferably to SEQ ID NO: 45, or g4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 45 and/or 250, preferably of SEQ ID NO: 45, or

g5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 45 and/or 250, preferably of SEQ ID NO: 45 per primer, or

g6) a polynucleotide of g1), g2), g3), g4) and/or g5) being obtainable from a pamamycin producing organism, or

g7) a polynucleotide of at least two of g 1 ), g2), g3), g4), g5) and g6);

at least one of h 1 ), h2), h3), h4), h5), h6) or h7) as defined in

hi ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, 258, 384, 391 , 398, 405 and/or 412, preferably to SEQ ID NO: 53, 258 and/or 384, more preferred to SEQ ID NO: 53, or

h2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, 54, 55, 56, 57, 58, 384, 385, 386, 387, 388, 389, 391 , 392, 393, 394, 395, 396, 398, 399, 400, 401 , 402, 403, 405, 406, 407, 408, 409, 410, 412, 413, 414, 415, 416 and/or 417, preferably to SEQ ID NO: 53, 54, 55, 56, 57, 58, 384, 385, 386, 387, 388 and/or 389, more preferred to SEQ ID NO: 53, 54, 55, 56, 57 and/or 58, or

h3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , preferably to SEQ ID NO: 52, 257 and/or 383, more preferred to SEQ ID NO: 52, or

h4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , preferably of SEQ ID NO: 52, 257 and/or 383, more preferred of SEQ ID NO: 52, or

h5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 52, 257, 383, 390, 397, 404 and/or 41 1 , preferably of SEQ ID NO: 52, 257 and/or 383, more preferred of SEQ ID NO: 52 per primer, or

h6) a polynucleotide of hi), h2), h3), h4) and/or h5) being obtainable from a pamamycin producing organism, or

h7) a polynucleotide of at least two of h 1 ), h2), h3), h4), h5) and h6);

at least one of i 1 ) , i2), i3), i4), i5), i6) or i7) as defined in

i1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60 and/or 265, preferably to SEQ ID NO: 60, or

12) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, 61 , 62, 63, 64, 65, 265, 266, 267, 268, 269 and/or 270, preferably to

SEQ ID NO: 60, 61 , 62, 63, 64 and/or 65, or

13) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 59 and/or 264, preferably to SEQ ID NO: 59, or

i4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 59 and/or 264, preferably of SEQ ID NO: 59, or

15) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 59 and/or 264, preferably of

SEQ ID NO: 59 per primer, or

16) a polynucleotide of i 1 ) , i2), i3), i4) and/or i5) being obtainable from a

pamamycin producing organism, or

17) a polynucleotide of at least two of i 1 ) , i2), i3), i4), i5) and i6);

j) at least one of j1), j2), j3), j4), j5), j6) or j7) as defined in

j1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67 and/or 272, preferably to SEQ ID NO: 67, or

j2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, 68, 69, 70, 71 , 72, 272, 273, 274, 275, 276 and/or 277, preferably to SEQ ID NO: 67, 68, 69, 70, 71 and/or 72, or

j3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 66 and/or 271 , preferably to SEQ ID NO: 66, or

j4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 66 and/or 271 , preferably of SEQ ID NO: 66, or

j5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 66 and/or 271 , preferably of SEQ ID NO: 66 per primer, or

j6) a polynucleotide of j 1 ) , j2), j3), j4) and/or j5) being obtainable from a

pamamycin producing organism, or

j7) a polynucleotide of at least two of j 1 ) , j2), j3), j4), j5) and j6);

k) at least one of k1 ), k2), k3), k4), k5), k6) or k7) as defined in k1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74 and/or 279, preferably to SEQ ID NO: 74, or

k2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, 75, 76, 77, 78, 79, 279, 280, 281 , 282, 283 and/or 284, preferably to SEQ ID NO: 74, 75, 76, 77, 78 and/or 79, or

k3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID

NO: 73 and/or 278, preferably to SEQ ID NO: 73, or

k4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 73 and/or 278, preferably of SEQ ID NO: 73, or

k5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 73 and/or 278, preferably of SEQ ID NO: 73 per primer, or

k6) a polynucleotide of k1), k2), k3), k4) and/or k5) being obtainable from a pamamycin producing organism, or

k7) a polynucleotide of at least two of k1 ), k2), k3), k4), k5) and k6);

at least one of 11 ), I2), I3), I4), I5), I6) or I7) as defined

11 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 and/or 286, preferably to SEQ ID NO:

81 , or

12) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , 82, 83, 84, 85, 86, 286, 287, 288, 289, 290 and/or 291 , preferably to SEQ ID NO: 81 , 82, 83, 84, 85 and/or 86, or

13) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 80 and/or 285, preferably to SEQ ID NO: 80, or

14) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 80 and/or 285, preferably of

SEQ ID NO: 80, or

15) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 80 and/or 285, preferably of SEQ ID NO: 80 per primer, or

16) a polynucleotide of 11), I2), I3), I4) and/or I5) being obtainable from a

pamamycin producing organism, or 17) a polynucleotide of at least two of 11 ), I2), I3), I4), I5) and I6);

m) at least one of ml ), m2), m3), m4), m5), m6) or m7) as defined

ml ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88 and/or 293, preferably to SEQ ID NO:

88, or

m2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, 89, 90, 91 , 92, 93, 293, 294, 295, 296, 297 and/or 298, preferably to SEQ ID NO: 88, 89, 90, 91 , 92 and/or 93, or

m3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87 and/or 292, preferably to SEQ ID NO: 87, or

m4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 87 and/or 292, preferably of

SEQ ID NO: 87, or

m5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 87 and/or 292, preferably of SEQ ID NO: 87 per primer, or

m6) a polynucleotide of ml ), m2), m3), m4) and/or m5) being obtainable from a pamamycin producing organism, or

m7) a polynucleotide of at least two of ml ), m2), m3), m4), m5) and m6);

n) at least one of n1), n2), n3), n4), n5), n6) or n7) as defined

n1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 95 and/or 300, preferably to SEQ ID NO: 95, or

n2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:

95, 96, 97, 98, 99, 100, 300, 301 , 302, 303, 304 and/or 305, preferably to

SEQ ID NO: 95, 96, 97, 98, 99 and/or 100, or

n3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 94 and/or 299, preferably to SEQ ID NO: 94, or

n4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 94 and/or 299, preferably of

SEQ ID NO: 94, or

n5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 94 and/or 299, preferably of SEQ ID NO: 94 per primer, or n6) a polynucleotide of n1), n2), n3), n4) and/or n5) being obtainable from a pamamycin producing organism, or

n7) a polynucleotide of at least two of n 1 ), n2), n3), n4), n5) and n6);

o) at least one of o1 ), o2), o3), o4), o5), 06) or o7) as defined in

o1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102 and/or 307, preferably to SEQ ID NO: 102, or

02) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:

102, 103, 104, 105, 106, 107, 307, 308, 309, 310, 31 1 and/or 312, preferably to SEQ ID NO: 102, 103, 104, 105, 106 and/or 107, or

03) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 101 and/or 306, preferably to SEQ ID NO: 101 , or

04) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 101 and/or 306, preferably of SEQ ID NO: 101 , or

05) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 101 and/or 306, preferably of SEQ ID NO: 101 per primer, or

06) a polynucleotide of o1), o2), o3), o4) and/or o5) being obtainable from a pamamycin producing organism, or

o7) a polynucleotide of at least two of o1 ), o2), o3), o4), o5) and 06);

p) at least one of p1 ), p2), p3), p4), p5), p6) or p7) as defined in

p1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109 and/or 314, preferably to SEQ ID NO: 109, or

p2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, 1 10, 1 1 1 , 1 12, 1 13, 1 14, 314, 315, 316, 317, 318 and/or 319, preferably to SEQ ID NO: 109, 1 10, 1 1 1 , 1 12, 1 13 and/or 1 14, or p3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 108 and/or 313, preferably to SEQ ID NO: 108, or

p4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 108 and/or 313, preferably of SEQ ID NO: 108, or

p5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 108 and/or 313, preferably of

SEQ ID NO: 108 per primer, or

p6) a polynucleotide of p1), p2), p3), p4) and/or p5) being obtainable from a pamamycin producing organism, or

p7) a polynucleotide of at least two of p1 ), p2), p3), p4), p5) and p6);

at least one of q 1 ), q2), q3), q4), q5), q6) or q7) as defined in

q1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%,

86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or

100% identical to SEQ ID NO: 1 16 and/or 321 , preferably to SEQ ID NO:

1 16, or

q2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17, 1 18, 1 19, 120, 121 , 321 , 322, 323, 324, 325 and/or 326, preferably to SEQ ID NO: 1 16, 1 17, 1 18, 1 19, 120 and/or 121 , or q3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15 and/or 320, preferably to SEQ ID NO: 1 15, or

q4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 1 15 and/or 320, preferably of SEQ ID NO: 1 15, or

q5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 and/or 320, preferably of SEQ ID NO: 1 15 per primer, or

q6) a polynucleotide of q1), q2), q3), q4) and/or q5) being obtainable from a pamamycin producing organism, or

q7) a polynucleotide of at least two of q 1 ), q2), q3), q4), q5) and q6);

at least one of r1 ), r2), r3), r4), r5), r6) or r7) as defined

r1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130 and/or 335, preferably to SEQ ID NO: 130, or

r2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, 131 , 132, 133, 134, 135, 335, 336, 337, 338, 339 and/or 340, preferably to SEQ ID NO: 130, 131 , 132, 133, 134 and/or 135, or r3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 129 and/or 334, preferably to SEQ ID NO: 129, or

r4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 129 and/or 334, preferably of SEQ ID NO: 129, or r5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 129 and/or 334, preferably of SEQ ID NO: 129 per primer, or

r6) a polynucleotide of r1), r2), r3), r4) and/or r5) being obtainable from a pamamycin producing organism, or

r7) a polynucleotide of at least two of r1 ), r2), r3), r4), r5) and r6);

s) at least one of s1 ), s2), s3), s4), s5), s6) or s7) as defined in

s1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or

100% identical to SEQ ID NO: 137 and/or 342, preferably to SEQ ID NO: 137, or

s2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, 138, 139, 140, 141 , 142, 342, 343, 344, 345, 346 and/or 347, preferably to SEQ ID NO: 137, 138, 139, 140, 141 and 142, or s3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 136 and/or 341 , preferably to SEQ ID NO: 136, or

s4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 136 and/or 341 , preferably of SEQ ID NO: 136, or

s5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 136 and/or 341 , preferably of

SEQ ID NO: 136 per primer, or

s6) a polynucleotide of s1), s2), s3), s4) and/or s5) being obtainable from a pamamycin producing organism, or

s7) a polynucleotide of at least two of s1 ), s2), s3), s4), s5) and s6);

and

t) at least one of t1 ), t2), t3), t4), t5), t6) or t7) as defined in

t1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123 and/or 327, preferably to SEQ ID NO: 123, or

t2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123, 124, 125, 126, 127, 128, 328, 329, 330, 331 , 332 and/or 333, preferably to SEQ ID NO: 123, 124, 125, 126, 127 and/or 128, or t3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %,

92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 122 and/or 326, preferably to SEQ ID NO: 122, or t4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 122 and/or 326, preferably of SEQ ID NO: 122, or

t5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 122 and/or 326, preferably of SEQ ID NO: 122 per primer, or

t6) a polynucleotide of t1), t2), t3), t4) and/or t5) being obtainable from a

pamamycin producing organism, or

t7) a polynucleotide of at least two of t1 ), t2), t3), t4), t5) and t6).

In one embodiment, the polynucleotides, expression cassettes, vectors and host cells comprise a polynucletotide of at least one of a) to t), being

a) a polynucletotide comprising

a1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, or

a2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 4, 5, 6, 7, 8, and/or 9, or

a3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3, or a4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 3, or

a5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 3 per primer, or

a6) a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a

pamamycin producing organism, or

a7) a polynucleotide of at least two of a1 ), a2), a3), a4), a5) and a6); b) a polynucletotide comprising

b1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 , or

b2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 1 , 12, 13, 14, 15 and/or 16, or

b3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 10, or b4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 10, or b5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 10 per primer, or

b6) a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a

pamamycin producing organism, or

b7) a polynucleotide of at least two of b1 ), b2), b3), b4), b5) and b6); a polynucletotide comprising

c1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18, or

c2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 18, 19, 20,

21 , 22 and/or 23, or

c3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17, or c4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 17, or

c5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 17 per primer, or

c6) a polynucleotide of c1), c2), c3), c4) and/or c5) being obtainable from a

pamamycin producing organism, or

c7) a polynucleotide of at least two of c1 ), c2), c3), c4), c5) and c6); a polynucletotide comprising

d1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, or

d2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 25, 26, 27,

28, 29 and/or 30, or

d3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 24, or d4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 24, or

d5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 24 per primer, or

d6) a polynucleotide of d1), d2), d3), d4) and/or d5) being obtainable from a

pamamycin producing organism, or

d7) a polynucleotide of at least two of d 1 ), d2), d3), d4), d5) and d6); a polynucletotide comprising

a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, or

a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32, 33, 34, 35, 36 and/or 37, or

a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31 , or a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 31 , or

a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 31 per primer, or

a polynucleotide of e1), e2), e3), e4) and/or e5) being obtainable from a pamamycin producing organism, or

a polynucleotide of at least two of e1 ), e2), e3), e4), e5) and e6); a polynucletotide comprising

f1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, or

f2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 39, 40, 41 ,

42, 43 and/or 44, or

f3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 38, or f4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 38, or

f5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 38 per primer, or

f6) a polynucleotide of f1), f2), f3), f4) and/or f5) being obtainable from a

pamamycin producing organism, or

f7) a polynucleotide of at least two of f1 ), f2), f3), f4), f5) and f6); a polynucletotide comprising

g1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46, or

g2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 46, 47, 48, 49, 50 and/or 51 , or

g3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 45, or g4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 45, or

g5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 45 per primer, or

g6) a polynucleotide of g1), g2), g3), g4) and/or g5) being obtainable from a

pamamycin producing organism, or

g7) a polynucleotide of at least two of g 1 ), g2), g3), g4), g5) and g6); a polynucletotide comprising

hi ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, or

h2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 53, 54, 55,

56, 57 and/or 58, or

h3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 52, or h4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 52, or

h5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 52 per primer, or

h6) a polynucleotide of hi), h2), h3), h4) and/or h5) being obtainable from a

pamamycin producing organism, or

h7) a polynucleotide of at least two of h 1 ), h2), h3), h4), h5) and h6); a polynucletotide comprising

11 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, or

12) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, 61 , 62, 63, 64 and/or 65, or

13) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 59, or

14) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 59, or

15) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 59 per primer, or

i6) a polynucleotide of i 1 ) , i2), i3), i4) and/or i5) being obtainable from a pamamycin producing organism, or

i7) a polynucleotide of at least two of i1 ), i2), i3), i4), i5) and i6); j) a polynucletotide comprising

j1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ I D NO: 67, or

j2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, 68, 69, 70, 71 and/or 72, or

j3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66, or j4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 66, or

j5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 66 per primer, or

j6) a polynucleotide of j 1 ) , j2), j3), j4) and/or j5) being obtainable from a pamamycin producing organism, or

j7) a polynucleotide of at least two of j 1 ) , j2), j3), j4), j5) and j6); k) a polynucletotide comprising

k1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, or

k2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 74, 75, 76,

77, 78 and/or 79, or

k3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 73, or k4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 73, or

k5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 73 per primer, or

k6) a polynucleotide of k1), k2), k3), k4) and/or k5) being obtainable from a

pamamycin producing organism, or

k7) a polynucleotide of at least two of k1 ), k2), k3), k4), k5) and k6); a polynucletotide comprising

11 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , or

12) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 81 , 82, 83, 84, 85 and/or 86, or

13) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 80, or

14) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 80, or

15) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 80 per primer, or

16) a polynucleotide of 11), I2), I3), I4) and/or I5) being obtainable from a pamamycin producing organism, or

17) a polynucleotide of at least two of 11 ), I2), I3), I4), I5) and I6); a polynucletotide comprising

a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, or

a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 88, 89, 90, 91 , 92 and/or 93, or

a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 87, or a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 87, or

a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 87 per primer, or

a polynucleotide of ml), m2), m3), m4) and/or m5) being obtainable from a pamamycin producing organism, or

a polynucleotide of at least two of ml ), m2), m3), m4), m5) and m6); a polynucletotide comprising

n1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 95, or

n2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 95, 96, 97, 98, 99 and/or 100, or

n3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 94, or n4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 94, or

n5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 94 per primer, or

n6) a polynucleotide of n1), n2), n3), n4) and/or n5) being obtainable from a

pamamycin producing organism, or

n7) a polynucleotide of at least two of n 1 ), n2), n3), n4), n5) and n6); a polynucletotide comprising

01 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102, or

02) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 102, 103, 104, 105, 106 and/or 107, or

03) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 101 , or

04) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 101 , or

05) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 101 per primer, or

06) a polynucleotide of o1), o2), o3), o4) and/or o5) being obtainable from a

pamamycin producing organism, or

07) a polynucleotide of at least two of o1 ), o2), o3), o4), o5) and 06); a polynucletotide comprising,

p1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, or

p2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 109, 1 10,

1 1 1 , 1 12, 1 13 and/or 1 14, or

p3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 108, or

p4) a polynucleotide which hybridises under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 108, or

p5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 108 per primer, or

p6) a polynucleotide of p1), p2), p3), p4) and/or p5) being obtainable from a

pamamycin producing organism, or

p7) a polynucleotide of at least two of p1 ), p2), p3), p4), p5) and p6); a polynucletotide comprising

q1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, or

q2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17,

1 18, 1 19, 120 and/or 121 , or

q3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15, or

q4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 1 15, or

q5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 per primer, or

q6) a polynucleotide of q1), q2), q3), q4) and/or q5) being obtainable from a

pamamycin producing organism, or

q7) a polynucleotide of at least two of q 1 ), q2), q3), q4), q5) and q6); a polynucletotide comprising

r1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, or

r2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 130, 131 ,

132, 133, 134 and/or 135, or

r3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 129, or

r4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 129, or

r5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 129 per primer, or r6) a polynucleotide of r1), r2), r3), r4) and/or r5) being obtainable from a pamamycin producing organism, or

r7) a polynucleotide of at least two of r1 ), r2), r3), r4), r5) and r6); a polynucletotide comprising

s1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, or

s2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 137, 138,

139, 140, 141 and 142, or

s3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 136, or

s4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 136, or

s5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 136 per primer, or

s6) a polynucleotide of s1), s2), s3), s4) and/or s5) being obtainable from a

pamamycin producing organism, or

s7) a polynucleotide of at least two of s1 ), s2), s3), s4), s5) and s6); a polynucletotide comprising

t1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123, or

t2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 123, 124,

125, 126, 127 and/or 128, or

t3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 122, or

t4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 122, or

t5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 122 per primer, or

t6) a polynucleotide of t1), t2), t3), t4) and/or t5) being obtainable from a

pamamycin producing organism, or

t7) a polynucleotide of at least two of t1 ), t2), t3), t4), t5) and t6); Preferably the polynucleotides, expression cassettes, vectors and host cells comprising a polynucleotide of at least one of a) to t)

a) at least one of a1), a2), a3), a4), a5), a6) or a7) as described above, and

b) at least one of b1), b2), b3), b4), b5), b6) or b7) as described above, and

c) at least one of c1 ), c2), c3), c4), c5), c6) or c7) as described above, and

d) at least one of d1), d2), d3), d4), d5), d6) or d7) as described above, and

e) at least one of e1), e2), e3), e4), e5), e6) or e7) as described above, and

f) at least one of f1), f2), f3), f4), f5), f6) or f7) as described above, and

g) at least one of g1), g2), g3), g4), g5), g6) or g7) as described above, and

h) at least one of hi), h2), h3), h4), h5), h6) or h7) as described above, and

i) at least one of i 1 ) , i2), i3), i4), i5), i6) or i7) as described above, and

j) at least one of j 1 ) , j2), j3), j4), j5), j6) or j7) as described above, and

k) at least one of k1 ), k2), k3), k4), k5), k6) or k7) as described above, and

I) at least one of 11), I2), I3), I4), I5), I6) or I7) as described above, and

m) at least one of ml ), m2), m3), m4), m5), m6) or m7) as described above, and n) at least one of n1), n2), n3), n4), n5), n6) or n7) as described above, and

o) at least one of o1), o2), o3), o4), o5), 06) or o7) as described above, and

P) at least one of p1 ), p2), p3), p4), p5), p6) or p7) as described above;

q) at least one of q1), q2), q3), q4), q5), q6) or q7) as described above, or

r) at least one of r1 ), r2), r3), r4), r5), r6) or r7) as described above, or

s) at least one of s1 ), s2), s3), s4), s5), s6) or s7) as described above, or

t) at least one of t1 ), t2), t3), t4), t5), t6) or t7) as described above

are recombinant polynucleotides or recombinant expression cassettes or vectors or host cells comprising such recombinant polynucleotides and/or recombinant expression cassettes.

Preferred embodiments define the polypeptides of a) to p), or the polypeptides of a) to t) as applicable, according to the criteria of a1) to p7) or a1) to t7), respectively, as described above, including preferred combinations and sequence identity regions of these criteria described in the different embodiments of the recombinant microorganisms disclosed above.

Preferred polynucleotides are

i) polynucleotides encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to at least one of SEQ ID NOs: 4, 1 1 , 18, 25, 32, 39, 46, 53, 60, 67, 74, 81 , 88, 95, 102 and 109.

ii) polynucleotides encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to at least one of the SEQ ID NOs listed in Tables 3 to 23.

iii) polynucleotides having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to at least one of SEQ ID NOs: 3, 10, 17, 24, 31 , 38, 45, 52, 59, 66, 73, 80, 87, 94, 101 and 108 iv) polynucleotides having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2

v) polynucleotides comprising expression cassettes having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a fragment of SEQ ID NO: 1 and/or 2.

Also preferred are expression cassettes, vectors and host cells comprising such

polynucleotides. In one embodiment these polynucleotides and expression cassettes are recombinant polynucleotides or recombinant expression cassettes.

Preferably, the vector referred to herein is suitable as a cloning vector or transformation vector, i.e. replicable in microbial systems or able to integrate polynucleotides into the genome of a microorganism. Also preferably, the vector of the present invention is an expression vector. Expression vectors comprise expression cassettes which enable the transcription and translation of the polynucleotides in the respective microorganism. The expression cassettes comprise a promoter and a terminator being operably linked to the polynucleotide coding for at least one polypeptide of the invention.

The polynucleotides encoding at least one of the polypeptides will preferably be adapted to the codon usage of the respective microorganism. Promoters, terminators and information about codon usage suitable to be used for a particular microorganism are known by a person skilled in the art.

Preferred promoters for microorganisms of the genus Streptomyces are the promoters known be a person skilled in the art. Examples of such promoters, but not excluding others, are:

the ermeE*p promoter (Schmitt-John and Engels, 1992, Promoter constructions for efficient expression in Streptomyces lividans, Appl Micobiol Biotechnol, 36, 493-498), having constitutive expression (SEQ ID NO: 192), the tipSp promoter (Holmes et al. 1993,

Autogenous transcriptional activation of a thiostrepton-induced gene in Streptomyces lividans, EMBO J, 12, 3183-3191), being inducible with thiostreptone,

the chi63p promoter (Ingram and Westpheling, 1995, ccrA1 : mutation in Streptomyces coelicolor that affects the control of catabolite repression, J Bacteriol, 177, 3579-3586) the gylCABp (Hindle et al, 1994, Substrate induction and catabolite repression of the Streptomyces coelicolor glycerol operon are mediated through the GylR protein, Mol Micobiol, 12, 737-745), being inducible with glycerol,

the mcrABp promoter (August et al., 1996, Inducible synthesis of the mitomycin C resistance gene product (MORA) from Streptomyces lavendulae, Gene, 175, 261 -267), being inducible with mitomycin C

Other suitable promoters can for example be found in Siegl, T et al. METABOLIC

ENGINEERING (2013) Volume:19, Pages:98-106, disclosing also sequence variants of SEQ ID NO: 192. The expression cassettes provided herein can be part of a vector generally used for cloning and multiplication in organisms, e.g. E. coli or yeast. The protein encoding polynucleotides provided herein can be part of expression vectors in order to produce the encoded polypeptides. A plurality of suitable expression vectors for different microorganisms are known in the art. Vectors for cloning and expression in E. coli are well known in the art. Examples of vectors for expression in S. cerevisiae comprise for example, but not excluding others, pYep Sec1 (Baldari 1987, Embo J. 6:229-234), pMFa (Kurjan 1982, Cell 30:933- 943), pJRY88 (Schultz 1987, Gene 54:1 13-123) and pYES2 (Invitrogen Corporation, San Diego, CA), pAG-1 , YEp6, YEp13 or pEMBLYe23

Vectors for cloning and expression in Streptomyces are for example, but not excluding others: plJ 101 , plJ364, plJ702, plJ361 , pTES, pTOS2, pSET152 and pOJ436

plJ6021 (Takano et al., 1995), plJ4123 (Takano et al., 1995), plJ4090, pMT3206, plTS107 (Kataoka et al., 1996), pANT849, plJ8600 (Sun et al., 1999), pPM927 (Smokvina et al., 1990), pHM6-1 1 (Motamedi et al., 1995), pMT3226, pKT02 (van Mellaert et al., 1998), pKC1064 (Kuhstoss abd Rao, 1991 ), pMCXpress (Ausust et al., 1996), pUWL201 (Doumith et al., 2000).

The examples of promoters and vectors mentioned above includes also their variants which have been constructed in the art. It will be clear to a person skilled in the art, that sequences located upstream of starting nucleotides of genes of SEQ ID NO: 1 will be able to provide promoter functions in Streptomyces, in particular in Streptomyces alboniger and Streptomyces albus.

It will also be clear to a person skilled in the art, that sequences located downstream of end point nucleotides of genes of SEQ ID NO: 1 will be able to provide terminator functions in Streptomyces, in particular in Streptomyces alboniger and Streptomyces albus.

Accordingly, a further embodiment of the invention are fragments of SEQ ID NO: 2 of about 2000, 1750, 1500, 1250, 1000, 750, 500, 300 or 250 nucleotides located immediately upstream of a starting nucleotide of a polypeptide encoding region of SEQ ID NO: 1 and having promoter function in Streptomyces, in particular in Streptomyces alboniger as well as fragments of SEQ ID NO: 2 of about 500, 300, or 250 nucleotides downstream of an stop codon of a polypeptide encoding region of SEQ ID NO: 2 and having terminator function in Streptomyces, in particular in Streptomyces alboniger, as well as recombinant expression cassettes, vectors and recombinant microorganisms comprising at least one of these fragments. Promoter regions can for example be found in SEQ ID NO: 2, upstream of the polypeptide encoding regions for the polypeptides having pamR2, pamW, pamC, pamF, pamA, pamD, pamK, pamX, pamS and pamR1 activity. Examples for such promoters are provided by SEQ ID NO: 143, 144 and 145. More information on promoter regions comprised by SEQ ID NO: 2 is provided in Example 9.

Thus, a preferred embodiment of the invention comprises

i) a polynucleotides which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 143, 144 or 145, or

ii) a polynucleotides which hybridise under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 143, 144 or 145, or

iii) a polynucleotides which are obtainable via PCR on genomic DNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of

SEQ ID NO: 143, 144 or 145 per primer, or

iv) a polynucleotide, of at least one of i), ii) or iii) and being obtainable from a pamamycin producing organism, or

v) a polynucleotide of at least one of i), ii) iii) or iv) and comprising promoter activity in Streptomyces alboniger.

Thus, a preferred embodiment of the invention comprises

i) recombinant polynucleotides, which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, identical to SEQ ID NO: 143,

144 or 145, or

ii) recombinant polynucleotides which hybridise under medium stringency hybridisation conditions, to a complement of SEQ ID NO: 143, 144 or 145, or

iii) recombinant polynucleotides which are at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a polynucleotide which is obtainable via PCR on genomic DNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 143, 144 or 145 per primer, or

iv) a polynucleotide of at least one of i), ii) or iii) and comprising promoter activity in

Streptomyces alboniger.

These promoters are preferably used in combination with a polypeptide having pamR1 activity. Preferably a polypeptide having pamR1 activity and enhancing expression from a promoter described by SEQ ID NO: 143, 144 or 145.

These promoter polypeptide combinations can be used to coordinate expression of genes in a given microorganism.

Hence, the invention comprises a system for coordinated gene expression in a

microorganism comprising

1 ) at least one expression cassette comprising a promoter being able to provide for gene expression in said microorganism and being operably linkted to

q1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, or

q2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17, 1 18, 1 19, 120 and/or 121 , or q3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15, or

q4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 1 15, or

q5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 per primer, or

q6) a polynucleotide of q1), q2), q3), q4) and/or q5) being obtainable from a

pamamycin producing organism, or

q7) a polynucleotide of at least two of q 1 ), q2), q3), q4), q5) and q6); and

2) at least one expression cassette comprising a promoter comprising

i) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 143, 144 or 145, or

ii) a polynucleotide which hybridise under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 143, 144 or 145, or

iii) a polynucleotide which is obtainable via PCR on genomic DNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive

polynucleotides of SEQ ID NO: 143, 144 or 145 per primer, or

iv) a polynucleotide of at least one of i), ii) or iii) and being obtainable from a

pamamycin producing organism, or

v) a polynucleotide of at least one of i), ii), iii), or iv) and comprising promoter

activity in Streptomyces alboniger.

Wherein the microorganism is preferably recombinant for at least this system for

coordinated gene expression, but may comprise further recombinant polynucleotides.

Further preferred embodiments are polynucleotides, expression cassettes, vectors and host cells comprising such promoters or terminators directly flanking at least one polypeptide encoding region of SEQ ID NO: 1 and vectors and host cells comprising a system for coordinated gene expression as described above.

Preferably, the recombinant microorganisms belong to the group of bacteria, preferably belonging to the group of Streptomyces or Amycolatopsis.

Preferred bacteria of the group Streptomyces or Amycolatopsis are selected from the group consisting of: Actinomadura madurae, Actinomyces violaceochromogenes, Kitasatosporia parulosa, Streptomyces achromogenes, Streptomyces acidiscabies, Streptomyces albus, Streptomyces ambofaciens, Streptomyces antibioticus, Streptomyces aureofaciens, Streptomyces aurantiacus, Streptomyces avermitilis, Streptomyces capreolus,

Streptomyces carcinostaticus, Streptomyces cervinus, Streptomyces clavuligerus,

Streptomyces coelicolor, Streptomyces coeruleorubidus, Streptomyces cyaneofuscatus, Streptomyces davawensis, Streptomyces diastaticus, Streptomyces fradiae, Streptomyces fulvissimus, Streptomyces ghanaensis, Streptomyces griseus,

Streptomyces griseoflavus, Streptomyces griseoviridus, Streptomyces hirsutus,

Streptomyces hygroscopicus, Streptomyces kanamyceticus, Streptomyces lavendulae, Streptomyces lividans, Streptomyces loidensis, Streptomyces natalensis, Streptomyces noursei, Streptomyces nodosum, Streptomyces olivaceus, Streptomyces platensis, Streptomyces peuceticus, Streptomyces pristinaespiralis, Streptomyces rimosus, Streptomyces roseosporus, Streptomyces sviceus, Streptomyces spectabilis,

Streptomyces tauricus, Streptomyces tendae, Streptomyces thioluteus, Streptomyces toxytricini, Streptomyces venezuelae, Streptomyces violaceoniger, Streptomyces violaceoruber, Streptomyces violaceus, Streptomyces viridochromogenes, Streptomyces violaceoruber and Streptomyces zaomyceticus.

In particular the Streptomyces species Streptomyces albus, Streptomyces auratus, Streptomyces avermitilis, Streptomyces coelicolor Streptomyces clavuligerus,

Streptomyces fulvissimus, Streptomyces ghanaensis, Streptomyces griseoflavus, Streptomyces hygroscopicus, Streptomyces lividans, Streptomyces pristinaespiralis, Streptomyces roseosporus, Streptomyces roseosporus, Streptomyces sviceus

Streptomyces viridochromogenes, Streptomyces violaceoruber. Preferred Streptomyces strains are: Streptomyces alboniger ATCC 12461, Streptomyces alboniger DSM 40043, Streptomyces alboniger DSM 40412, Streptomyces albus B24108, Streptomyces albus J 1074, Streptomyces albus DSM40313, Streptomyces avermitilis SUKA17 or Streptomyces avermitilis SUKA22, Streptomyces coelicolor DSM40233, Streptomyces kitasatoensis JCM5000, Streptomyces kitasatoensis JCM5001 Streptomyces lividans ATCC69444.

In particular preferred are microorganisms which already have the capacity to produce pamamycin such as, but not excluding others: Streptomyces alboniger ATCC 12461 Streptomyces alboniger DSM 40043, Streptomyces aurantiacus ATCC 19822 and

Streptomyces kitasatoensis JCM5001.

The polynucleotides and polypeptides described can be used to identify new pamamycin producing organisms or to identify microorganisms potentially capable to produce pamamycin.

Microorganisms potentially capable to produce pamamycin comprise one, several or most of the expression cassettes for polypeptides having pamC activity, pam G activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity, pamS activity, pamR1 activity, pamR2 activity, pamW activity or pamH activity, while one or more expression cassettes for polypeptides having pamC activity, pam G activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity, pamS activity are defective or missing, so that these microorganisms do not produce pamamycin.

Microorganisms potentially capable to produce pamamycin can be transformed into pamamycin producing organisms by providing them with functional expression cassettes to complement the defective or missing expression cassettes.

Accordingly, the invention includes methods to identify pamaycin producing microorganisms or microorganisms potentially capable to produce pamamycin comprising the steps of a) providing a microorganism,

b) isolating DNA and/or cDNA from such microorganism,

c) identifying polynucleotide sequences encoding polypeptides having at least one of pamC activity, pam G activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity, pamS activity, by

I) hybridizing the isolated DNA or RNA under medium or high stringent conditions with polynucleotide fragments of SEQ ID NO: 1 or 2 or both, or

II) performing PCR reactions with PCR-primer suitable to amplify fragments of SEQ ID NO: 1 or 2 or both

III) sequencing the isolated DNA and/or RNA and perform sequence comparisons with polynucleotide sequences of fragments of SEQ ID NO: 1 or 2 or both, d) selecting microorganisms which comprise sequences encoding polypeptides for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15 or all of the polynucleotide sequences of d), e) testing the selected microorganisms for production of pamamycin.

Preferably the microorganisms of step a) are also tested for the presence of polynucleotide sequences encoding polypeptides having at least one of pamR1 activity, pamR2 activity, pamW activity and pamH activity.

Polynucleotide sequences encoding polypeptides having at least one of pamC activity, pam G activity, pamF activity, pamA activity, pamB activity, pamD activity, pamE activity, pamO activity, pamK activity, pamJ activity, pamM activity, pamN activity, pamL activity, pamX activity, pamY activity, pamS activity, can be identified by any sutiable method known in the art to identify polynucleotides encoding certain polypeptides, including the hybridisation, PCR, RNA and analysis of sequenced genome information having been described herein.

Preferably the microorganisms of step a) are tested for the presence of 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15 polynucletotides of a) to t)

a) at least one of a1 ), a2), a3), a4), a5), a6) or a7) as described above, and

b) at least one of b1), b2), b3), b4), b5), b6) or b7) as described above, and

c) at least one of c1 ), c2), c3), c4), c5), c6) or c7) as described above, and

d) at least one of d 1 ), d2), d3), d4), d5), d6) or d7) as described above, and

e) at least one of e1), e2), e3), e4), e5), e6) or e7) as described above, and f) at least one of f1), f2), f3), f4), f5), f6) or f7) as described above, and

g) at least one of g 1 ), g2), g3), g4), g5), g6) or g7) as described above, and

h) at least one of hi), h2), h3), h4), h5), h6) or h7) as described above, and

i) at least one of i 1 ) , i2), i3), i4), i5), i6) or i7) as described above, and

j) at least one of j 1 ) , j2), j3), j4), j5), j6) or j7) as described above, and

k) at least one of k1 ), k2), k3), k4), k5), k6) or k7) as described above, and

I) at least one of 11), I2), I3), I4), I5), I6) or I7) as described above, and

m) at least one of ml ), m2), m3), m4), m5), m6) or m7) as described above, and n) at least one of n1), n2), n3), n4), n5), n6) or n7) as described above, and

o) at least one of o1), o2), o3), o4), o5), 06) or o7) as described above, and

p) at least one of p1), p2), p3), p4), p5), p6) or p7) as described above;

q) at least one of q 1 ), q2), q3), q4), q5), q6) or q7) as described above, or

r) at least one of r1 ), r2), r3), r4), r5), r6) or r7) as described above, or

s) at least one of s1 ), s2), s3), s4), s5), s6) or s7) as described above, or

t) at least one of t1 ), t2), t3), t4), t5), t6) or t7) as described above.

Preferably the methods to identify pamaycin producing microorganisms or microorganisms potentially capable to produce pamamycin comprise also the following steps iii) and iv) iii) selecting an microorganism comprising polynucleotides of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19 or all of a) to t)

iv) testing the selected microorganism for the production of at least one pamamycin, preferably testing for the production of pamamycin 607 or pamamycin 621 or both.

Preferably the microorganisms are selected in step iii) which comprise at least a

polynucleotide of i) and/or j), being

i) a polynucletotide comprising

i1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, or

i2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 60, 61 , 62, 63, 64 and/or 65, or

i3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%,

93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 59, or i4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 59, or

i5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 59 per primer, or

i6) a polynucleotide of i1), i2), i3), i4) and/or i5) being obtainable from a pamamycin producing organism, or

i7) a polynucleotide of at least two of i 1 ), i2), i3), i4), i5) and i6); j) a polynucletotide comprising

j1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, or

j2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67, 68, 69, 70, 71 and/or 72, or

j3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 66, or j4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 66, or

j5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 66 per primer, or

j6) a polynucleotide of j 1 ) , j2), j3), j4) and/or j5) being obtainable from a pamamycin producing organism, or

j7) a polynucleotide of at least two of j 1 ) , j2), j3), j4), j5) and j6); The invention provides also for methods to produce a recombinant microorganism for the production of pamamycin comprising the steps of:

i) providing genomic DNA or cDNA or genomic DNA information of a microorganism; ii) testing the genomic DNA or cDNA or genomic DNA information for the presence of at least one polynucleotide for each one of a) to p);

a) at least one of a1), a2), a3), a4), a5), a6) or a7) as described above, and b) at least one of b1), b2), b3), b4), b5), b6) or b7) as described above, and c) at least one of c1 ), c2), c3), c4), c5), c6) or c7) as described above, and d) at least one of d 1 ), d2), d3), d4), d5), d6) or d7) as described above, and e) at least one of e1), e2), e3), e4), e5), e6) or e7) as described above, and f) at least one of f 1 ), f2), f3), f4), f5), f6) or f7) as described above, and

g) at least one of g1), g2), g3), g4), g5), g6) or g7) as described above, and h) at least one of hi), h2), h3), h4), h5), h6) or h7) as described above, and i) at least one of i 1 ) , i2), i3), i4), i5), i6) or i7) as described above, and

j) at least one of j 1 ) , j2), j3), j4), j5), j6) or j7) as described above, and

k) at least one of k1 ), k2), k3), k4), k5), k6) or k7) as described above, and

I) at least one of 11), I2), I3), I4), I5), I6) or I7) as described above, and

m) at least one of ml ), m2), m3), m4), m5), m6) or m7) as described above, and n) at least one of n1), n2), n3), n4), n5), n6) or n7) as described above, and o) at least one of o1), o2), o3), o4), o5), 06) or o7) as described above, and p) at least one of p1 ), p2), p3), p4), p5), p6) or p7) as described above;

iii) providing the microorganism with expression cassettes comprising polynucleotides of those one of a) to p), of which no presence could be confirmed in step ii); iv) testing the microorganism of step iii) for the production of at least one pamamycin, preferably testing for the production of pamamycin 607 or pamamycin 621 or both.

The invention provides also methods to enhance the production of pamamycin, preferably the production of pamamycin 607 or pamamycin 621 or both, in a microorganism comprising i) at least one polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2, preferably 2,

ii) at least one polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2, preferably 2, and comprising at least one expression cassette for each one of a) to p)

a) at least one of a1 ), a2), a3), a4), a5), a6) or a7) as described above, and b) at least one of b1), b2), b3), b4), b5), b6) or b7) as described above, and c) at least one of c1), c2), c3), c4), c5), c6) or c7) as described above, and d) at least one of d 1 ), d2), d3), d4), d5), d6) or d7) as described above, and e) at least one of e1), e2), e3), e4), e5), e6) or e7) as described above, and f) at least one of f 1 ), f2), f3), f4), f5), f6) or f7) as described above, and

g) at least one of g1), g2), g3), g4), g5), g6) or g7) as described above, and h) at least one of hi), h2), h3), h4), h5), h6) or h7) as described above, and i) at least one of i 1 ) , i2), i3), i4), i5), i6) or i7) as described above, and

j) at least one of j 1 ) , j2), j3), j4), j5), j6) or j7) as described above, and

k) at least one of k1), k2), k3), k4), k5), k6) or k7) as described above, and

I) at least one of 11), I2), I3), I4), I5), I6) or I7) as described above, and

m) at least one of ml ), m2), m3), m4), m5), m6) or m7) as described above, and n) at least one of n1), n2), n3), n4), n5), n6) or n7) as described above, and

0) at least one of o1), o2), o3), o4), o5), 06) or o7) as described above, and p) at least one of p1), p2), p3), p4), p5), p6) or p7) as described above;

iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments

comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of

1) or ii), if the fragments are combined.

by providing the microorganism with at least one expression cassette comprising a promoter being able to provide for gene expression in said microorganism and being operably linkted to

q1 ) a polynucleotide encoding a polypeptide which is at least 80%, 82%, 84%, 86%,

88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, or

q2) a polynucleotide encoding a polypeptide which is at least 90%, 91 %, 92%, 93%,

94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 16, 1 17,

1 18, 1 19, 120 and/or 121 , or

q3) a polynucleotide which is at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1 15, or

q4) a polynucleotide which hybridises under medium stringency hybridisation

conditions, to a complement of SEQ ID NO: 1 15, or

q5) a polynucleotide which is obtainable via PCR on genomic DNA or cDNA of a pamamycin producing organisms using primer pairs of at least 15 consecutive polynucleotides of SEQ ID NO: 1 15 per primer, or

q6) a polynucleotide of q1), q2), q3), q4) and/or q5) being obtainable from a

pamamycin producing organism, or

q7) a polynucleotide of at least two of q 1 ), q2), q3), q4), q5) and q6).

Preferred microorganisms used in the methods to identify pamaycin producing

microorganisms or microorganisms potentially capable to produce pamamycin or used in methods to produce a recombinant microorganism for the production of pamamycin or used in methods to enhance the production of pamamycin are microorganisms of from the genus Streptomyces alboniger, Streptomyces ambofaciens, Streptomyces antibioticus,

Streptomyces aurantiacus, Streptomyces caviscabies, Streptomyces cyaneofuscatus, Streptomyces griseus, Streptomyces kitasatoensis, Streptomyces olivaceus, Streptomyces tauricus, Streptomyces tendae, Streptomyces viridochromogenes and Streptomyces zaomyceticus.

In one embodiment of the invention, the recombinant microorganism is produced by transforming microorganism with at least one of the gene clusters described by Figure 1 comprising at least one polynucleotide of i), ii) or iii) as described below.

i) a polynucleotide having at least 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2, preferably 2,

ii) a polynucleotide having at least 70%, 72%, 74%, 76%, 78%, 80%, 82%, 84%, 86%, 88%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1 and/or 2, preferably 2, and comprising at least one expression cassette for each one of a) to p)

a) at least one of a1), a2), a3), a4), a5), a6) or a7) as described above, and b) at least one of b1), b2), b3), b4), b5), b6) or b7) as described above, and c) at least one of c1 ), c2), c3), c4), c5), c6) or c7) as described above, and d) at least one of d 1 ), d2), d3), d4), d5), d6) or d7) as described above, and e) at least one of e1), e2), e3), e4), e5), e6) or e7) as described above, and f) at least one of f1), f2), f3), f4), f5), f6) or f7) as described above, and

g) at least one of g1 ), g2), g3), g4), g5), g6) or g7) as described above, and h) at least one of hi), h2), h3), h4), h5), h6) or h7) as described above, and i) at least one of i 1 ) , i2), i3), i4), i5), i6) or i7) as described above, and

j) at least one of j 1 ) , j2), j3), j4), j5), j6) or j7) as described above, and

k) at least one of k1 ), k2), k3), k4), k5), k6) or k7) as described above, and I) at least one of 11), I2), I3), I4), I5), I6) or I7) as described above, and m) at least one of ml ), m2), m3), m4), m5), m6) or m7) as described above, and n) at least one of n1), n2), n3), n4), n5), n6) or n7) as described above, and o) at least one of o1), o2), o3), o4), o5), 06) or o7) as described above, and

P) at least one of p1), p2), p3), p4), p5), p6) or p7) as described above;

iii) two or more fragments of the polynucleotides of i) or ii) wherein the fragments

comprise functional expression cassettes for one or more of the polypeptides of a) to p) and wherein the fragments cover at least the whole sequence of a polynucleotide of i) or ii), if the fragments are combined.

Preferably the expression cassettes of the gene clusters will comprise promoters and terminator sequences which lead to high expression in the respective microorganism.

Preferably also the polynucleotide encoding sequences of the gene clusters will be codon optimized for the respective microorganism. The gene clusters will not need to be transformed as one single element, but may be transformed in several pieces, which may or may not integrate at different locations in the genome. Usually all pieces of the gene cluster comprise complete expression cassettes, so that none of the expression cassettes is destroyed in case pieces of the gene cluster are integrated at different locations in the genome of the microorganism.

Preferably the microorganisms transformed with such clusters are microorganisms of from the groups of Streptomyces or Amycolatopsis, including the preferred species of both groups as described above.

Preferred embodiments of the methods to identify pamaycin producing microorganisms or microorganisms potentially capable to produce pamamycin and of the methods to produce a recombinant microorganism and of the methods to enhance the production of pamamycin define the polypeptides of a) to p), or the polypeptides of a) to t) as applicable, according to the criteria of a1) to p7) or al ) to t7), respectively, as described above, including preferred combinations and sequence identity regions of these criteria described in the different embodiments of the recombinant microorganisms disclosed above.

Preferably, the methods to Preferred microorganisms used in the methods to identify pamaycin producing microorganisms or microorganisms potentially capable to produce pamamycin or used in methods to produce a recombinant microorganism for the production of pamamycin or used in methods to enhance the production of pamamycin comprise an additional step wherein the recombinant microorganism is tested for the production of pamamycin. The recombinant microorganisms can be tested for the production of pamamycin, by culturing the recombinant microorganism under conditions which allow for the production of pamamycin and analysing the recombinant microorganism or the culture medium or analysing the recombinant microorganism and the culture medium for the presence of pamamycin. The invention comprises also the use of the microorganisms discloced herein in a method for the production of pamamycin, as well as the use of any one of the polynucleotides, expression cassettes, vectors disclosed herein in a method to produce pamamycin, in a method to identify pamaycin producing microorganisms or microorganisms potentially capable to produce pamamycin, or in a method to produce a recombinant microorganism for the production of pamamycin, or in a method to enhance the production of pamamycin.

All references cited in this specification are herewith incorporated by reference with respect to their entire content and the content specifically mentioned in this specification.

EXAMPLES

1. Culturing of Streptomyces alboniger DSM 40043

A 300 ml culture flasks with 4 baffles, comprising 100 ml of preculture medium (NL410) was inoculated with spores of Streptomyces alboniger DSM 40043 and incubated at 28 °C and 120 rpm for 48h.

The main culture was grown in 1 I shaking flasks with 2 baffles at 28 °C and 120 rpm for 120 h and inoculated with 4 % of preculture. The main culture was grown in either PPM medium or in SSG medium.

N410 medium:

Glucose 10 g

Glycerol 10 g

Oatmeal 5 g

Soymeal 10 g

Yeastextract 5 g

Bacto Casaminoacids 5 g

CaC03

Tap water liter, pH 7,0

SGG medium:

Starch soluble 10 g

Glucose 10 g

Glycerol 10 g

Cornsteep Powder 2,5 g

Bacto Peptone 5 g

Yeastextract 2 g

NaCI i g

CaC03 3 g

Tap water liter, pH 7,3

PPM medium: Maltose monohydrate 15 g

Polypeptone 10 g

Yeastextract 3 g

NaCI 3 g

CaC03 2 g

Distilled water ad 1 liter,

Example 2. Identification and analysis of pamamycin biosynthetic gene cluster.

The genome of Streptomyces alboniger ATCC 12461 (DSM40043) was sequenced and assembled into 10 scaffolds of different size. The pamamycin biosynthesis gene cluster was identified based on the similarity of its predicted biosynthetic pathway to the biosynthesis of the macrotetrolide antibiotic nonactin produced by Streptomyces griseus ETH A7796.

However, the proposed steps of pamamycin biosynthesis involve also an aminotransferase and methyltransferase(s) which are not comprised by the nonactin biosynthesis gene cluster. In order to compensate for this, sequences of aminotransferase and

methyltransferase(s) were also used as targets in genome mining.

Within the genome of S. alboniger two distinct regions containing ketosynthase (KS) genes similar to polyketidsynthases (PKS) being involved in nonactin biosynthesis have been identified. The region of scaffold 1 , (554075-580246 bp) is located in close proximity to a PKSI-NRPS gene cluster. This region contains 7 KS genes, two of which (orf0731 (46% identities), orf0740 (48% identities)) show a high degree of homology to the NonK ketosynthase of the nonactin biosynthesis gene cluster. Other KS (orf 0730, 0732, 0734, 0736, 0738) resemble KS-a of PKSII. Interestingly, this cluster contains an ACP gene (orf0729) that was found to be not needed in nonactin assembly. The identified region also contains genes encoding three dehydrogenase (orf0737, 0741 , 0742), acyl-CoA-transferase (orf0733), aminotransferase (orf0745), two methyltransferase (orf0746, 0757), hydrolase (orf07444), transporter (orf0727), enoyl CoA dehydratase/isomerase (orf0747) and two regulatory genes of the TetR (orf0726) and LuxR (orf0749) families.

A S. alboniger cosm\d library was constructed on a shuttle Streptomyces - E. col) cosmid vector pOJ436 and screened by hybridization with two probes (5' probe being described in SEQ ID NO: 146 and a 3' probe - being described in SEQ ID NO: 147) covering 31458 bp of the S. alboniger chromosome containing the identified region with all genes predicted to be involved in pamamycin biosynthesis and 2 and 4 kb of flanking sequence. Five library clones (R1 , R2, R3, R4, and R5) giving strong signals with both probes were identified and end-sequenced. The resulting sequences proved that the identified cosmids contained the desired region of the S. a/bon/ger chromosome.

Example 3. Deletion of the pmm0734 PKS encoding gene (pamD) in the native pamamycin gene cluster of S. alboniger.

A plasmid for the deletion of the pmm0734 gene was constructed based on cloning two 1.6 kb regions of the pamD polypeptide ecoding region in the vector pKC1 139. A Streptomycin resistance marker gene (aadA) was inserted between the cloned regions flanking the pamD polypeptide encoding regions, so that these flanking regions could be used as " recombination arms" in order to replace the pmm0734 region with the aadA gene in the chromosome of S. alboniger. Successful gene replacement was confirmed by PCR. S. alboniger strains comprising this gene replacement were grown in culture media and tested for pamamycin production. All clones comprising the gene replacement failed to produce spores and pamamycin.

Example 4. Heterologous expression of cosmids containing the pamamycin gene cluster.

The five cosmids (R1 to R5) identified in Example 2 were transformed in the heterologous host Streptomyces albus J 1074. Exconjugants for the cosmids R1 , R2, R3, and R4 were obtained and tested for pamamycin production. The exconjugants containing cosmids R1 , R2 and R3 produced pamamycins, of wich exconjugants comprising the cosmid R2 (S. albus R2) had the highest production level. Recombinant strain S. albus R2 was found also to produce several pamamycins of which the major compounds were pamamycins 607, 621 and 635. Culture media supernatant of S. albus R2 comprised about 3.5 mg/L pamamycin 607, which is about twice as much as the wild type strain of S. alboniger. While the wild type strain of S. alboniger contained more pamamycin in the produced biomass (see Figure 25a), so that the overall production of pamamycin 607 was higher in S. alboniger DSM 40043 than in the heterologous host S. albus comprising the R2 cosmid.

Example 5. Overexpression of the the pamamycin gene cluster in Streptomyces alboniger DSM 40043 and comparison of pamamycin production with wildtype Streptomyces alboniger DSM 40043 and heterologous host S. alboniger comprising cosmid R2.

Cosmid R2 having been obtained in Example 2 was transformed in the homologous host Streptomyces alboniger DSM 40043 in order to produce a new strain having an additional copy of the pamamycin cluster. A recombinant strain of Streptomyces alboniger DSM 40043 comprising the R2 plasmid (S. alboniger + R2) as well as a wildtype Streptomyces alboniger DSM 40043 (S. alboniger) and a strain of Streptomyces albus comprising a R2 cosmid as described in Example 4 (S. albus + R2) were tested for pamamycin production in two different media (PPM medium and SGG medium as described in Example 1 ). Culture medium supernatant of PPM medium (PPM KF) and culture medium supernatant of SGG medium (SGG KF) as well as produced biomass of each medium " PMM Mycel" and " SGG Mycel" of each of these strains were separately analysed for their content of pamamycin (see Figure 25b). Again, S. albus + R2 secreted more pamamycin in the medium in comparison to S. alboniger and S. alboniger + R2. While both variants of S. alboniger comprised more pamamycin in the biomass and had a higher overall production of pamamycin. All three strains produced more pamamycin in the SGG medium than in the PPM medium. Example 6. Production of R2 cosmid variants comprising deletions of pam Genes.

Deletion of pam genes within the R2 cosmid was performed with the use of homologous recombination. For this purpose a set of primers with 39 bp of flanking region of target gene and fragment complementary to the resistance gene cassette were used to generate the disruption PCR product. Primers used in this work and respective primers to check for the gene exchange with the resistance marker via PCR are listed in Table 24. Mutated cosmids were transformed into S. albus. Chromosomal DNA from these strains was purified and the presence of the mutated R2 variants was verified.

Table 24:

Gene deletion Cloning primers Check primers

pamC SEQ ID NOs 148, 149 SEQ ID NOs 150, 151 pamG SEQ ID NOs 152, 153 SEQ ID NOs 154, 155 pamF SEQ ID NOs 156, 157 SEQ ID NOs 158, 159 pamA SEQ ID NOs 160, 161 SEQ ID NOs 162, 163 pamB SEQ ID NOs 164, 165 SEQ ID NOs 166, 167 pamO SEQ ID NOs 168, 169 SEQ ID NOs 170, 171 pamJ SEQ ID NOs 172, 173 SEQ ID NOs 174, 175 pamL SEQ ID NOs 176, 177 SEQ ID NOs 178, 179 pamX SEQ ID NOs 180, 181 SEQ ID NOs 182, 183 pamY SEQ ID NOs 184, 185 SEQ ID NOs 186, 187 pamR1 SEQ ID NOs 188, 189 SEQ I D NOs 190, 191 S. albus strains comprising the R2 cosmid having a deleted pamF, pamB, pamJ, pamL, pamX, pamY or pamR1 gene failed to produce pamamycin.

S. albus strains comprising the R2 cosmid having a deleted pamA or pamC gene produced far less pamamycin than S. albus strains comprising the complete R2 cosmid. This indicates that S. albus comprises endogenous polypeptides having a low pamA or pamC activity, respectively.

Example 7. Generation of Streptomyces albus strains comprising the R2 cosmid and an additional pamR1 gene.

In order to increase the overall yield of pamamycin in S. albus comprising the R2 cosmid, the pmm0749 gene encoding pamR1 was cloned into the pTOS integrative vector under control of its native and the strong constitutive heterologous promoter p21 (SEQ ID NO: 193), respectively. Resulting plasmids pTOS0749BX (native promoter) and pTOS0749P21 (P21 p) were introduced into S. albus J 1074 and the vector backbone was removed by expression of the Dre recombinase. The cosmid R2 was introduced into the obtained strains and production of pamamycin was evaluated. In the case of S. albus 0749P21 containing the R2 cosmid, about 1.3 fold overall increase in pamamycin accumulation could be observed when compared to S. albus J 1074.

Example 8. Generation of Streptomyces alboniger strains comprising an additional pamR1 gene. A wildtype S. alboniger strain and a S. alboniger strain comprising the R2 cosmid were provided with an additional copy of the pamR1 gene via plasmid pTOSR1 P21 (P21 promoter driving pamR1 expression) and via plasmid pTOSRI BX (native promoter driving pamR1 expression). The resulting strains were tested for their pamamycin production in PPM and SGG medium of Example 1.

Introduction of additional copies of pamR1 into S. alboniger caused up to a 2.5 fold increase of pamamycin 607 production (see Figure 26).

Example 9. Transcriptional analysis of the pamamycin cluster in the heterologous host S. albus.

In order to investigate the transcriptional organization of the pam gene cluster, RNAseq analysis experiments were performed using a S. albus strain comprising the R2 cosmid. The results showed that the pamamycin gene cluster is organized in several transcribed units. The two major operons starts with two divergent promoters located within the pamF and pamA intergenic region.

The promoter of the pamA gene is transcribing a large multicistronic mRNA that includes the genes pamA, B, D, E, O. The predicted transcription start is located 48 bp away from the ATG start codon of the pamA gene. The other promoter transcribes a large

multicistronic mRNA that includes genes K, J, M , N, L and H.

Genes pamF and pamG are trascribed in one mRNA which is under transcriptional control of a promoter upstream of the pamF encoding region. Its transcription initiation point is located at about 33 bp away from the start codon of pamF.

The pamC gene is transcribed from its own promoter with the transcription initiation position located 21 bp away from the start codon.

The pamX and pamY genes are also transcribed as an operon with their transcription initiation site located 69 bp upstream from the pamX start codon.

The pamS gene transcription is initiated 42 bp upstream from the start codon.

The pamR1 transcription start is initiated 1 1 to 13 bp away from its predicted start codon.

The pamW gene comprises its own promoter and produces a mRNA coding only for the pamW polypeptide.

A further promoter is located upstream of the pamR2 encoding region. The produced mRNA lacks a 5' -untranslated region and its transcription initiates directly at the

translation starting point. Example 10. Activity of the pamA, pamD and pamR1 promoter in the homologous host Streptomyces alboniger and in the heterologous host Streptomyces albus.

In order to test the promoter activity of the pamA promoter (SEQ ID NO: 143), the pamD promoter (SEQ I D NO: 144) and the pamR1 promoter (SEQ ID NO: 145), recombinant plasmids were constructed in which the gusA reporter gene with an ATG start codon is operatively linked to the respective promoters of the pamA, pamD and pamR1 gene. The resulting plasmids were introduced into S. albus and S. alboniger and GusA activity was monitored. The relative GusA activity is shown in Table 25. Table 25:

promoter GusA activity in S. GusA activity in S.

alboniger DSM 40043 albus J 1074

pamA + ( ⁺ )

pamR1 + ++

pamD ++ +++

(+) represents weak activity

+ represents activity

++ represents strong activity

+++ represents very strong activity

Example 1 1 : Deletion of the pmm0731 encoding gene (pamF) in the heterologous expression host Streptomyces albus J 1074

The deletion of the gene pmm0731 (pamF) was performed using the Red/ET recombination technology. A hygromycin resistance marker gene (hyg) flanked by modfied attB sites of the 0C31 bacteriophage was excised from the patt-hyg plasmid (see Myronovskyi, M., B.

Rosenkranzer, and A. Luzhetskyy, Iterative marker excision system. Appl Microbiol Biotechnol, 2014. 98(10): p. 4557-70) as a BamHI/Xbal fragment using the primers 0731 DF (SEQ I D NO: 194) and 0731 DR (SEQ ID NO: 194). The obtained PCR product was used to replace the gene pmm0731 (pamF) within the R2 cosmid with the hygromycin resistance gene. Successful gene replacement was confirmed by PCR. The mutated cosmids were introduced into Streptomyces albus J 1074. Since the R2 cosmid contains the int gene for 0C31 the hyg resistance marker was immediately removed during cosmid integration into the genome of the host strain, thus generating a markerless mutation of the desired gene. The strain was grown in culture media and tested for pamaycin production. The deletion of pmm0731 (pamF) caused a complete cessation of hydroxy acid L, whereas hydroxy acid S was still produced. Example 12: Deletion of the pmm0730 encoding gene (pamG) in the heterologous expression host Streptomyces albus J 1074

The deletion of the gene pmm0730 (pamG) was performed using the Red/ET recombination technology. A hygromycin resistance marker gene (hyg) flanked by modfied attB sites of the 0C31 bacteriophage was excised from the patt-hyg plasmid (see Myronovskyi, M., B., et al in Example 1 1 ) as a BamHI/Xbal fragment using the primers 0730DelHygMF (SEQ ID NO: 196) and 0730DelHygMR (SEQ ID NO: 197). The obtained PCR product was used to replace the gene pmm0730 (pamG) within the R2 cosmid with the hygromycin resistance gene. Successful gene replacement was confirmed by PCR. The mutated cosmids were introduced into Streptomyces albus J 1074. Since the R2 cosmid contains the int gene for 0C31 the hyg resistance marker was immediately removed during cosmid integration into the genome of the host strain, thus generating a markerless mutation of the desired gene. The strain was grown in culture media and tested for pamaycin production. The deletion of pmm0730 (pamG) caused a complete cessation of hydroxy acid L, whereas hydroxy acid S was still produced.

Example 13: Deletion of the pmm0740 encoding gene (pamJ) in the heterologous expression host Streptomyces albus J 1074

The deletion of the gene pmm0740 (pamJ) was performed using the Red/ET recombination technology. A hygromycin resistance marker gene (hyg) flanked by modfied attB sites of the 0C31 bacteriophage was excised from the patt-hyg plasmid (see Myronovskyi, M., B., et al in Example 1 1 ) as a BamHI/Xbal fragment using the primers 0740DF (SEQ ID NO: 198) and 0740DR (SEQ ID NO: 199). The obtained PCR product was used to replace the gene pmm0740 (pamJ) within the R2 cosmid with the hygromycin resistance gene. Successful gene replacement was confirmed by PCR. The mutated cosmids were introduced into Streptomyces albus J 1074. Since the R2 cosmid contains the int gene for 0C31 the hyg resistance marker was immediately removed during cosmid integration into the genome of the host strain, thus generating a markerless mutation of the desired gene. The strain was grown in culture media and tested for pamaycin production. The deletion of pmm0740 (pamJ) caused an accumulation of hydroxy acids S and L.

Example 14: Deletion of the pmm0745 encoding gene (pamX) in the heterologous expression host Streptomyces albus J 1074

The deletion of the gene pmm0745 (pamX) was performed using the Red/ET recombination technology. A hygromycin resistance marker gene (hyg) flanked by modfied attB sites of the 0C31 bacteriophage was excised from the patt-hyg plasmid (see Myronovskyi, M., B., et al in Example 1 1 ) as a BamHI/Xbal fragment using the primers 0745hygDF (SEQ ID NO: 200) and

0745hygDR (SEQ ID NO: 201 ). The obtained PCR product was used to replace the gene pmm0745 (pamX) within the R2 cosmid with the hygromycin resistance gene. Successful gene replacement was confirmed by PCR. The mutated cosmids were introduced into Streptomyces albus J 1074. Since the R2 cosmid contains the int gene for 0C31 the hyg resistance marker was immediately removed during cosmid integration into the genome of the host strain, thus generating a markerless mutation of the desired gene. The strain was grown in culture media and tested for pamaycin production. The deletion of pmm0745 (pamX) caused an accumulation of hydroxy acids S and K. Example 15: Deletion of the pmm0746 encoding gene (pamY) in the heterologous expression host Streptomyces albus J 1074

The deletion of the gene pmm0746 (pamY) was performed using the Red/ET recombination technology. A hygromycin resistance marker gene (hyg) flanked by modfied attB sites of the 0C31 bacteriophage was excised from the patt-hyg plasmid (see Myronovskyi, M., B., et al in Example 1 1 ) as a BamHI/Xbal fragment using the primers 0746DelF3 (SEQ ID NO: 202) and 0746DelR3 (SEQ ID NO: 203). The obtained PCR product was used to replace the gene pmm0746 (pamY) within the R2 cosmid with the hygromycin resistance gene. Successful gene replacement was confirmed by PCR. The mutated cosmids were introduced into Streptomyces albus J 1074. Since the R2 cosmid contains the int gene for 0C31 the hyg resistance marker was immediately removed during cosmid integration into the genome of the host strain, thus generating a markerless mutation of the desired gene. The strain was grown in culture media and tested for pamaycin production. The deletion of pmm0746 (pamY) caused an accumulation of hydroxy acids S and K.

Example 16: Deletion of the pmm0738 encoding gene (pamK) in the heterologous expression host Streptomyces albus J 1074

The deletion of the gene pmm0738 (pamK) was performed using the Red/ET recombination technology. A hygromycin resistance marker gene (hyg) flanked by modfied attB sites of the 0C31 bacteriophage was excised from the patt-hyg plasmid (see Myronovskyi, M., B., et al in Example 1 1 ) as a BamHI/Xbal fragment using the primers 0738DelHygMF (SEQ ID NO: 204) and 0738DelHygMR (SEQ ID NO: 205). The obtained PCR product was used to replace the gene pmm0738 (pamY) within the R2 cosmid with the hygromycin resistance gene. Successful gene replacement was confirmed by PCR. The mutated cosmids were introduced into Streptomyces albus J 1074. Since the R2 cosmid contains the int gene for 0C31 the hyg resistance marker was immediately removed during cosmid integration into the genome of the host strain, thus generating a markerless mutation of the desired gene. The strain was grown in culture media and tested for pamaycin production. The deletion of pmm0738 (pamK) caused an accumulation of hydroxy acids S and L.

Example 17: Deletion of the pmm0743 encoding gene (pamL) in the heterologous expression host Streptomyces albus J 1074

The deletion of the gene pmm0743 (pamL) was performed using the Red/ET recombination technology. A hygromycin resistance marker gene (hyg) flanked by modfied attB sites of the 0C31 bacteriophage was excised from the patt-hyg plasmid (see Myronovskyi, M., B., et al in Example 1 1 ) as a BamHI/Xbal fragment using the primers 0743DhygMF2 (SEQ ID NO: 206) and 0743DhygMR2 (SEQ ID NO: 207). The obtained PCR product was used to replace the gene pmm0743 (pamL) within the R2 cosmid with the hygromycin resistance gene. Successful gene replacement was confirmed by PCR. The mutated cosmids were introduced into Streptomyces albus J 1074. Since the R2 cosmid contains the int gene for 0C31 the hyg resistance marker was immediately removed during cosmid integration into the genome of the host strain, thus generating a markerless mutation of the desired gene. The strain was grown in culture media and tested for pamaycin production. The deletion of pmm0743 (pamL) caused an accumulation of hydroxy acids S and L.

Example 18. Complementation of S. albus strains having pamA, pamB, pamC, pamJ, pamL, pamO, pamX and pamY knockouts with pamA, pamB, pamC, pamJ, pamL, pamO, pamX and pamY genes

The complementation of the knockout strains with deleted pmm genes was performed with pmm genes cloned into the pUWLFLPhyg (see H. J. Kwon, W. C. Smith, A. J. Scharon, S. H. Hwang, M. J. Kurth, B. Shen, Science 2002, 297, 1327-1330. M. Fedoryshyn, L. Petzke, E. Welle, A. Bechthold, A. Luzehtskyy, Gene 2008, 419, 43-47) vector under the control of the ErmE* promoter (SEQ ID NO: 192). Transformed S. albus strains containing the respective complementation construct were selected by hygromycin resistance.

S. albus strains comprising the corresponding gene complementation construct were grown in culture media and tested for pamamycin production. All strains produced pamamycin.

Example 19. Complementation of S. albus pamB and pamO knockouts strains with nonT and nonO genes of Streptomyces fulvissimus DSMZ 40767.

The complementation of the strain with the deleted pamB gene and pamO gene was performed with the homologous nonT gene (SEQ ID NO: 348) and the homologous nonO gene (SEQ ID NO: 383) of Streptomyces fulvissimus DSMZ 40767. The nonT and nonO gene, respectively, were cloned into the pUWLFLPhyg vector under the control of the ErmE* promoter (SEQ ID NO: 192). Transformed S. albus strains containing the

complementation construct were selected by hygromycin resistance.

S. albus strains comprising the nonT gene or nonO gene were grown in culture media and tested for pamamycin production. Both strains produced pamamycin.

Example 20. Feeding experiment with L-chain and S-chain

A S. albus strain containing the pamJ, pamK and pamL genes was cultivated and fed with a mixture of compound S-chain and compound L-chain. The culture medium was tested for pamamycin production. The strain produced pamaycin.