REFERENCE TO SEQUENCE LISTING

The present application contains a sequence listing in computer readable form, which is incorporated by reference.

FIELD OF INVENTION

The invention relates to the production of oxygenated diterpenoid compounds in recombinant cells, such as yeast cells. The oxygenated diterpenoid compounds are useful as intermediate or final compounds in the synthesis of useful bioactive compounds for use in e.g. pharmaceutical treatment of diseases such as cancer. The invention further relates to genes, enzymes and cells, such as yeast cells, particularly suited for production of such compounds.

BACKGROUND FOR THE INVENTION

Terpenes is a diverse group of compounds generated from a basic 5-carbon structure, isoprene (2-methyl-1 ,3-butadiene). Diterpenes are compounds having a 20-carbon structure, generated by the action of an enzyme, diterpene synthase, that converts the compound geranyl-geranyl-diphosphate (GGPP) into a diterpene structure, which can by further modified forming a vast range of diterpene or diterpenoid compounds.

Diterpenes, diterpenoids, derivatives thereof are widely used, e.g. as pharmaceuticals, cosmetics, nutraceuticals, flavors, fragrances and pesticides. Methods for increasing the production of these compounds in natural or engineered cells are abundant in the art.

The Chinese medicinal plant, Tripterygium wilfordii, is known to produce several sesquit- erpenoids, diterpenoids and triterpenoids with potential pharmacological properties, including the diterpenoid compounds triptonide and triptolide. Triptolide, an oxygenated diterpenoid compound, and derivatives thereof, has been identified as potential valuable pharmacological compounds and is under investigation as immunosuppressant and for treatment of cancer. Triptolide may further be used in treatment against COVID-19. Triptonide may be useful as male contraceptive agent.

Using engineered microorganisms for producing valuable molecules from renewable feedstock is a desirable alternative from conventional means of production. However, achieving economically viable yield, titers and productivity is a major roadblock towards industrialisation. N L Hansen et al (2017) in The Plant Journal 2017, 89, 429-441, described a diterpene synthase capable of converting GGPP into the dipterpene, miltiradiene, which is a pre- curser for triptolide. The findings were confirmed by P Su et al (2018) in The plant Jour- nal 2018, 50-65: and by J Guo et al. PNAS 2013, 110, 12108-12113. The complete pathway for converting miltiradiene into other diterpenoid compounds, such as triptolide, has not yet been elucidated. Cytochrome P450 enzymes (CYPs) are involved in the biosynthesis of terpenoids, and for many cytochrome P450 enzymes nothing is known with respect to the substrate they are acting on, which compounds they are generating, or their role in the biosynthe- sis of specific compounds. US 20190270971A1 discloses methods for increasing productivity of microbial host cells that functionally express p450 enzymes. The document describes how P450 genes can be modified in order to improve performance in microorganisms, such as yeast, and it mentions that co-expression with cytochrome P450 reductase can be ben- eficial to improve the yield. It is mentioned that triptolide may be the subject of P450 chemistry, but the document does not provide any link between triptolide and any spe- cific P450 enzymes or cytochromes. CN 108395997A describes yeast with increased GGPP production. The yeast is trans- formed with different diterpene synthases and P450 enzymes to synthesize diterpenoid compounds. The scientist team behind this patent is also behind more patents and pa- tent applications disclosing the synthesis of different di- and triterpenoid compounds using suitable terpene synthases and P450 enzymes e.g. CN 108866029 (friedelin), CN107058419 (Kauren-type) and WO 2020029564 (Fridelin and amyrins). CN 110747178A describes the P450 gene TwCYP728B70 as encoding a Cytochrome P450 enzyme having a role in triptolide synthesis. SHORT DESCRIPTION OF THE INVENTION The inventors have solved the problem of providing an improved method of producing oxygenated diterpenoid compounds, such as triptophenolide, triptonide and triptolide. In a first aspect the invention relates to a method for producing an oxygenated diterpe- noid compound is disclosed, the method comprising the steps of: a. providing a host cell capable of producing miltiradiene and/or dehydro- abietadiene; b. transforming the host cell with a first gene encoding an enzyme having cytochrome P450 activity; c. growing the transformed cell under conditions leading to expression of the transformed gene; whereby the oxygenated diterpenoid compound is formed; wherein: the first gene encoding an enzyme having cytochrome P450 activity encodes a pol- ypeptide comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 1 (TwCYP82D274v1), SEQ ID NO: 2 (TwCYP82D274v2), SEQ ID NO: 74 (TwCYP82D274v3) or SEQ ID NO: 75 (TwCYP82D274v4), or the mature polypeptide thereof. In a second aspect, the invention relates to methods for producing oxygenated diterpe- noid compound, and comprises transforming the host cell with the first gene encoding an enzyme having cytochrome P450 activity and further with a second gene encoding a second enzyme having cytochrome P450 activity and with a third gene encoding a third enzyme having cytochrome P450 activity wherein: the second gene encoding an enzyme having cytochrome P450 activity encodes a polypeptide comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 3 (TwCYP71BE85) or the mature polypeptide thereof; and the third gene encoding an enzyme having cytochrome P450 activity encodes a polypeptide comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 4 (TwCYP71BE85) or the mature polypeptide thereof. In a third aspect, the invention relates to methods for producing oxygenated diterpenoid compound, wherein said method comprises that the host cell is transformed with the first, second and third genes encoding enzymes having cytochrome P450 activities, and fur- ther with a fourth gene encoding a fourth enzyme having cytochrome P450 activity; wherein: the fourth gene encoding a fourth enzyme having cytochrome P450 activity en- codes a polypeptide comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% se- quence identity to SEQ ID NO: 5 (TwCYP82D213v1) or to SEQ ID NO: 76 (TwCYP82D213v2), or the mature polypeptide thereof. According to the invention the useful oxygenated diterpenoid compounds, triptophe- nolide, triptonide or triptolide, may be provided. The invention further relates to polypeptides, polynucleotides, plasmids and expression constructs as well as recombinant host cells useful in the methods of the invention. SHORT DESCRIPTION OF THE DRAWINGS Figure 1 shows LCMS profiles of extracts from N. benthamiana leafs expressing mil- tirdiene biosynthesis genes and selected T. wilfordii CYPs. TwCYPs was co-expressed with CfDXS, CfGGPPS, CfTPS1 and CfTPS3. "3xSTD" rep- resents a LCMS run of a sample of three mixed authentic standards: Triptolide, trip- tonide, and triptophenolide. Solid lines represent ion chromatograms at range m/z 280- 380. Dashed lines (- ----- -) represent extracted ion chromatograms at m/z 313.1800±0.015 which corresponds to the parental ion of triptophenolide [M+H]. Dashed lines (- - -) represents extracted ion chromatograms at m/z 359.1490±0.015 which cor- responds to the parental ion of triptonide [M+H]. LC protocol 1 used. For more details see example 1. Figure 2 shows LCMS profiles of extracts of genetically engineered Saccharomyces cerevisiae (S. cerevisiae) strain. In the background strain (-), genes encoding the diterpene biosynthetic enzymes SPGG- PPS7, CftTPS1, CftTPS3 and TwCPR1 were integrated into genome of wild type S. cerevisiae. In the TwCYP82D274v1 strain, the diterpene biosynthetic enzymes were ex- pressed with TwCYP82D274v1, resulting in the formation of compound (3) identified as 14-OH-dehydroabietadiene marked in grey. LC method1 was used for the analysis. For more details see example 3. Figure 3 shows the ¹ H NMR spectrum of 14-OH-dehydroabietadiene in CDCl ₃ at 599.85 MHz.. For more details see example 4. Figure 4 shows the ¹³ C NMR spectrum of 14-OH-dehydroabietadiene in CDCl ₃ at 150.83 MHz. For more details see example 4. Figure 5 shows LCMS profiles of extracts of yeast having denoted gene combinations genome integrated. All yeast strains have genome integrated spGGPPs7, CftTPS1, CftTPS3 and TwCPR1. "0.5ppm 3xSTD" represents a LCMS run of a sample of three mixed authentic standards: Triptolide, triptonide and triptophenolide. Non-dashed lines represent ion chromato- grams at range m/z 280-380. Dashed lines represents extracted ion chromatograms at m/z 359.1490±0.015 which corresponds to the parental ion of triptonide ([M+H] ⁺ ). LC protocol 2 used. For more details see example 5. Figure 6 shows co-expression of TwCYPs and different variants of B5 proteins isolated from Tripterygium wilfordii Levels of triptophenolide, triptonide and 14-OH-dehydroabietadiene quantified from cul- tures of engineered S. cerevisiae strains. Each column represents an engineered yeast strain and their output of selected compounds. The genes integrated into each of the individual strains are denoted in the lower panel. Quantification was based on the areas of the peaks representing each of the compounds of interest. Individual scales applies for each of the compounds. For more details see example 5. Figure 7 shows relative quantity (bars) of key intermediates in the proposed biosyn- thetic pathway of triptonide, when established in vivo, via heterologous gene expres- sion in N. benthamiana (panel A and D) and S. cerevisiae (panel B and E, strains listed in table 3). Gene expression is indicated by black squares to the left, while relative quantity is indicated by bars (average of 3-4 biological replicates; black diamond squares) with white- and grey fill color distinguishing expression and no expression of Twb5#1, respectively. Error bars represent standard deviation. “DiTPSs” reflects Cft- TPS1 and CftTPS3. In quantification of peak areas, the signature mass tolerance was ±0.1m/z for GCMS (miltiradiene and 14-OH-dehydroabietadiene) and ±0.005m/z for LCMS (all other compounds). Panel C: A hypothetized biosynthetic pathway from milti- radiene to triptonide in vivo in N. benthamiana and S. cerevisiae that include a Wagner- Meerwein rearrangement reaction to account for a methyl shift of C-19 or C-18 to C-3 in the abietane carbon backbone. Figure 8 shows accumulation over 7 days of triptophenolide and triptonide produced with yeast strain NVJ8.15 when grown in bioreactor. Level of triptonide (solid black line) and triptophenolide (dotted black line) shows absolute quantity (ppm, w/v) in samples of the culture taken each day. Biomass was quantified by absorbance at 600nm (grey dotted line). Figure 9 shows yeast strains expressing the genes needed for triptonide biosynthesis, but with genes variants substituting TwCYP82D274v1 or TwCYP82D213v1, pertain the ability to produce triptophenolide (panel A) and triptonide (panel B) and results in simi- lar LCMS profiles (panel C). Genes present in the engineered strains are represented by the black squares. Panel A and B: Bars represent the average relative quantity (2-3 biological replicates, crosses) with error bars showing std. error. From left to right, bars represent yeast strains: NVJ10-1, NVJ10-3, NVJ10-6, NVJ10-8 (see table 3). Panel C: EICs (m/z 280-360) of LCMS analyzed yeast cultures. From top and down pairs of chromatograms represent yeast strains NVJ10-1, NVJ10-3, NVJ10-6 and NVJ10-8. SHORT DESCRIPTION OF THE SEQUENCES SEQ ID NO: 1 shows the amino acid sequence of a cytochrome P450 enzyme derived from T. wilfordii. The enzyme is also known as TwCYP82D274v1. SEQ ID NO: 2 shows the amino acid sequence of a cytochrome P450 enzyme derived from T. wilfordii. The enzyme is also known as TwCYP82D274v2. SEQ ID NO: 2 differs from SEQ ID NO: 1 in only three positions, and it is therefore assumed that SEQ ID NO: 1 and SEQ ID NO: 2 represent different alleles of the same gene. SEQ ID NO: 3 shows the amino acid sequence of a cytochrome P450 enzyme derived from T. wilfordii. The enzyme is also known as TwCYP71BE85. SEQ ID NO: 4 shows the amino acid sequence of a cytochrome P450 enzyme derived from T. wilfordii. The enzyme is also known as TwCYP71BE86. SEQ ID NO: 5 shows the amino acid sequence of a cytochrome P450 enzyme derived from T. wilfordii. The enzyme is also known as TwCYP82D213v1. SEQ ID NO: 6 shows the amino acid sequence of a cytochrome P450 enzyme derived from T. wilfordii. The enzyme is also known as TwCYP82D217. SEQ ID NO: 7 shows the amino acid sequence of a cytochrome P450 enzyme derived from T. wilfordii. The enzyme is also known as TwCYP82D275. SEQ ID NO: 8 shows the amino acid sequence of a cytochrome B5 enzyme derived from T. wilfordii. This enzyme is also known as TwB5#1. SEQ ID NO: 9 shows the amino acid sequence of a cytochrome P450 reductase enzyme derived from T. wilfordii. This enzyme is also known as TwCPR1. SEQ ID NO: 10-66 show PCR primers as further described in Example 2. SEQ ID NO: 67 shows the amino acid sequence of a diterpene synthase TPS1 derived from Plectranthus barbatus. The enzyme is also known as CfTPS1. SEQ ID NO: 68 shows the amino acid sequence of a diterpene synthase TPS3 derived from Plectranthus barbatus. The enzyme is also known as CfTPS3. SEQ ID NO: 69 shows the amino acid sequence of a terpene synthase TPS9, derived from T. wilfordii. The enzyme is also known as TwTPS9. SEQ ID NO: 70 shows the amino acid sequence of a terpene synthase derived from T. wilfordii. The enzyme is also known as TwTPS27. SEQ ID NO: 71 shows the amino acid sequence of a copalyl diphosphate synthase CPS1 derived from Salvia miltiorrhiza. The enzyme is also known as SmCPS. SEQ ID NO: 72 shows the amino acid sequence of a miltiradiene synthase KSL1 derived from Salvia miltiorrhiza. The enzyme is also known as SmKSL. SEQ ID NO: 73 shows the amino acid sequence of a geranyl geranyl diphosphate syn- thase derived from Synechococcus sp. The enzyme is also known as SpGGPPs7v1. SEQ ID NO: 74 shows the amino acid sequence of a cytochrome P450 enzyme derived from T. wilfordii. The enzyme is also known as TwCYP82D274v3. SEQ ID NO: 75 shows the amino acid sequence of a cytochrome P450 enzyme derived from T. wilfordii. The enzyme is also known as TwCYP82D274v4. SEQ ID NO: 76 shows the amino acid sequence of a cytochrome P450 enzyme derived from T. wilfordii. The enzyme is also known as TwCYP82D213v2. SEQ ID NO: 77 shows the truncated amino acid sequence of a diterpene synthase TPS1 derived from Plectranthus barbatus. The amino acid sequence was truncated to remove a transit peptide. The enzyme is also known as CftTPS1. SEQ ID NO: 78 shows the truncated amino acid sequence of a diterpene synthase TPS3 derived from Plectranthus barbatus. The amino acid sequence was truncated to remove a transit peptide. The enzyme is also known as CftTPS3. SEQ ID NO: 79 shows the amino acid sequence of a DXS enzyme derived from Plec- tranthus barbatus. The enzyme is also known as CfDXS. SEQ ID NO: 80 shows the amino acid sequence of a truncated HMGR enzyme derived from S. cerevisiae. The enzyme is also known as SctHMGR. SEQ ID NO: 81 shows the amino acid sequence of a geranyl geranyl diphosphate syn- thase derived from Synechococcus sp. The enzyme is also known as SpGGPPs7v2. DETAILED DESCRIPTION OF THE INVENTION According to a first aspect of the invention, a method for producing an oxygenated diterpenoid compound is disclosed, the method comprising the steps of: a. providing a host cell capable of producing miltiradiene and/or dehydro- abietadiene; b. transforming the host cell with a first gene encoding an enzyme having cytochrome P450 activity; c. growing the transformed cell under conditions leading to expression of the transformed gene; whereby the oxygenated diterpenoid compound is formed; wherein: the first gene encoding an enzyme having cytochrome P450 activity encodes a pol- ypeptide comprising or consisting of SEQ ID NO:1 or an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity more preferred at least 98% sequence identity to SEQ ID NO: 1 (TwCYP82D274V1) or the mature pol- ypeptide thereof. The polypeptide having SEQ ID NO: 1 is a preferred example of a first gene having cytochrome P450 activity, the polypeptides having SEQ ID NO: 2, SEQ ID NO: 74 and SEQ ID NO: 75 are other examples of such a polypeptide. Thus, the enzyme encoded by the first gene has the ability to convert miltiradiene and/or dehydroabietadiene into 14-OH-dehydroabietadiene by inserting an OH group in position 14 of the diterpene skeleton of miltiradiene. In some embodiments, the synthesis of 14-OH-dehydroabietadiene takes place via the compound 14-OH-miltiradiene that subsequently is converted into 14-OH-dehydro- abietadiene. However, the invention is not limited to any particular mechanism for con- verting miltiradiene into 14-OH-dehydroabietadiene. Using the method according to the first aspect of the invention leads to the formation of the oxygenated diterpenoid compound, 14-OH-dehydroabietadiene,

that is a useful intermediate in the synthesis of oxygenated diterpenoid compounds of pharmaceutical use, including well known compounds such as triptophenolide, triptonide and triptolide. In a second aspect of the invention, the method step b. comprises transforming the host cell with the first gene encoding an enzyme having cytochrome P450 activity and further with a second gene encoding a second enzyme having cytochrome P450 activity and a third gene encoding a third enzyme having cytochrome P450 activity wherein: the second gene encoding an enzyme having cytochrome P450 activity encodes a polypeptide comprising or consisting of SEQ ID NO:4 or an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 4 (TwCYP71BE86) or the mature poly- peptide thereof; and the third gene encoding an enzyme having cytochrome P450 activity encodes a polypeptide comprising or consisting of SEQ ID NO: 3 or an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 3 (TwCYP71BE85) or the mature poly- peptide thereof. In this second aspect, the host cell preferably further produces the oxygenated diterpe- noid compound, triptophenolide, (3bR,9bS)-6-hydroxy-9b-methyl-7-propan-2-yl- 3,3b,4,5,10,11-hexahydronaphtho[2,1-e]isobenzofuran-1-one.

Triptophenolide is a valuable compound that has been identified as an antiandrogen. In addition, it may be useful as a starting point for further modifications leading to further bioactive compounds. In a preferred embodiment of the second aspect of the invention, the host cell is further transformed with a fifth gene encoding a polypeptide having cytochrome B5 activity and comprising or consisting of SEQ ID NO:8 or an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 8 (TwB5#1) or the mature polypeptide thereof. It has surprisingly been found that expressing the polypeptide having cytochrome B5 ac- tivity in the same cell that expresses the first, second and third genes encoding enzymes having cytochrome P450 activity, leads to a significantly higher production of the oxy- genated diterpenoid compound. The production is increased at least 50% compared with the production of a similar cell without the polypeptide having cytochrome B5 activity, preferably increased at least 100%, preferably at least 200% or even more. In a third aspect of the invention, the host cell is transformed with the first, second and third genes encoding enzymes having cytochrome P450 activities, and further with a fourth gene encoding a fourth enzyme having cytochrome P450 activity; wherein: the fourth gene encoding a fourth enzyme having cytochrome P450 activity en- codes a polypeptide comprising or consisting of SEQ ID NO:5 or an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, pref- erably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 5 (TwCYP82D213v1) or the mature polypeptide thereof. The polypeptide having SEQ ID NO: 5 is a preferred exam- ple of a fourth gene having cytochrome P450 activity; the polypeptide having SEQ ID NO: 76 is another example of such a polypeptide. In the third aspect of the invention, the transformed eukaryotic cell preferably produces the oxygenated diterpenoid compound, triptonide. The compound triptonide has been reported to have a strong inhibition activity in cancers (Fulu Dong et al 2019, The Prostate, Volume 19, issue 11, pages 1284-1293). The com- pound is also useful as male contraceptive agent. Further, the compound is useful as a starting point for further modifications leading to further bioactive compounds. In a preferred embodiment of the third aspect of the invention, the host cell is further transformed with a fifth gene encoding a polypeptide having cytochrome B5 activity and comprising or consisting of SEQ ID NO:8 or an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 8 (TwB5#1) or the mature polypeptide thereof. It has surprisingly been found that expressing the polypeptide having cytochrome B5 ac- tivity in the same cell that expresses the first, second, third and fourth genes encoding enzymes having cytochrome P450 activity, leads to a significantly higher production of the oxygenated diterpenoid compound. The production is increased at least 50% com- pared with the production of a similar cell without the polypeptide having cytochrome B5 activity, preferably increased at least 100%, preferably at least 200% or even more. In a further preferred embodiment of the third aspect of the invention, the host cell is further transformed with a sixth gene encoding a fifth enzyme having cytochrome P450 activity and/or a seventh gene encoding a sixth enzyme having cytochrome P450 activity wherein: the sixth gene encoding a fifth enzyme having cytochrome P450 activity encodes a polypeptide comprising or consisting of SEQ ID NO:6 or an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 6 (TwCYP82D217) or the mature poly- peptide thereof ; and the seventh gene encoding a sixth enzyme having cytochrome P450 activity en- codes a polypeptide comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% se- quence identity to SEQ ID NO: 7 (TwCYP82D275) or the mature polypeptide thereof. It has surprisingly been found that expressing the sixth and/or seventh genes encoding enzymes having cytochrome P450 activity, leads to a higher production of the oxygen- ated diterpenoid compound. Preferably, the production is increased at least 10% com- pared with the production of a similar cell without sixth and/or seventh genes encoding enzymes having cytochrome P450 activity, preferably increased at least 20%, even more preferred at least 50% or even more. The first, second, third, fourth, fifth, sixth and seventh gene may be comprised in one or more nucleic acid molecules, such as one or more heterologous nucleic acids. The het- erologous nucleic acid encoding the first enzyme having cytochrome P450 activity may herein be referred to as the “first heterologous nucleic acid”. The heterologous nucleic acid encoding the second enzyme having cytochrome P450 activity may herein be re- ferred to as the “second heterologous nucleic acid”. The heterologous nucleic acid en- coding the third enzyme having cytochrome P450 activity may herein be referred to as the “third heterologous nucleic acid”. The heterologous nucleic acid encoding the fourth enzyme having cytochrome P450 activity may herein be referred to as the “fourth het- erologous nucleic acid”. The heterologous nucleic acid encoding the fifth enzyme hav- ing cytochrome P450 activity may herein be referred to as the “fifth heterologous nu- cleic acid”. The heterologous nucleic acid encoding the sixth enzyme having cyto- chrome P450 activity may herein be referred to as the “sixth heterologous nucleic acid”. The heterologous nucleic acid encoding the enzyme having cytochrome B5 activity may herein be referred to as the “seventh heterologous nucleic acid”. This does not im- ply that the recombinant host cell must comprises seven heterologous nucleic acids in total; in some embodiments, the cell comprises only one or more of the first, second, third, fourth, fifth, sixth and seventh heterologous nucleic acids. The oxygenated diterpenoid compounds produced according to the methods of the in- vention may be further modified by biological or chemical synthesis. In connection with this, biological synthesis is understood as a method where the host cell comprising the genes of the invention is further provided with one of more additional genes encoding further enzymes having the capability of modifying the oxygenated diterpenoid com- pounds produced according to the methods of the invention. Chemical modification of the oxygenated diterpenoid compounds produced according to the methods of the invention may be performed directly on the culture broth before re- covery of the oxygenated diterpenoid compounds or it may be performed on the recov- ered oxygenated diterpenoid compounds. Reduction of triptonide to triptolide can be achieved by organic synthesis. An example of such synthesis is the reduction by a nucleophilic attack by a hydride on C-14 ketone. For this reaction Sodium borohydride is a suitable agent for catalyzing this reaction at neutral pH in the appropriate solvent e.g. water or MetOH. In one preferred embodiment, triptonide produced according to the methods of the in- vention is converted into the compound triptolide, that is reported to be an immunosup- pressant and is under investigation for use in cancer therapy.

The host cell The host cell capable of producing miltiradiene and/or dehydroabietadiene may in prin- ciple be any such cell. The cell may be a cell that naturally produces miltiradiene and/or dehydroabietadiene or it may be a cell that has been engineered to produce one or both of these compounds. It is believed that miltiradiene, at least under some circumstances, may be spontaneously converted into dehydroabietadiene, and the invention may therefore be performed using a cell producing miltiradiene that is spontaneously converted into dehydroabietadiene or it may be performed in a cell comprising an enzyme that facilitates the conversion of miltiradiene to dehydroabietadiene (see a J. Zi, et al., Organic & Biomolecular Chemistry 2013, 11, 7650-7652). The synthesis of miltiradiene in general begins with the formation of GGPP. GGPP may be synthesized by condensation of one dimethylallyl pyroophosphate (DMAP) molecule and three isopentenyl pyrophosphate (IPP) molecules and is typically catalyzed by a geranylgeranyl diphosphate synthase e.g. the SpGGPPs7 enzyme derived from Syn- echococcus sp.; and having the amino acid sequence shown in SEQ ID NO: 73 or SEQ ID NO: 81. GGPP is converted into miltiradiene by action of a diterpene synthase or by the combined action of two or more diterpene synthases, e.g. a combination of two diterpene syn- thases, CfTPS1 and CfTPS3, or CftTPS1 as set forth in SEQ ID NO: 77 and CftTPS3 as set forth in SEQ ID NO: 78, derived from Plectranthus barbatus and having the amino acid sequences of SEQ ID NO: 67 and 68; a combination of two diterpene synthases, TwTPS9 and TwTPS27, derived from T. wilfordii and having the amino acid sequences of SEQ ID NO: 69 and 70; or a combination of a copalyl diphosphate synthase, SmCPS derived from Salvia miltiorrhiza and having the amino acid sequence of SEQ ID NO: 71, and a miltiradiene synthase, SmKSL derived from Salvia miltiorrhiza and having the amino acid sequence of SEQ ID NO: 72. One preferred way to provide a host cell producing miltiradiene and/or dehydroabietadi- ene is selecting a host cell producing GGPP and transforming it with a diterpene syn- thase catalyzing the transformation of GGPP into miltiradiene. Alternatively, a host cell that have been genetically engineered to produce GGPP may be used as a starting point. Techniques for transforming a host cell with a diterpene synthase catalyzing the trans- formation of GGPP into miltiradiene is known in the prior art, e.g. in N L Hansen et al (2017) in The Plant Journal 2017, 89, 429-441 (Incorporated herein by reference), P Su et al (2018) in The plant Journal 2018, 50-65 and in : J. Guo, et al., Proceedings of the National Academy of Sciences 2013, 110, 12108-12113, and the procedures and meth- ods disclosed in these publications are also useful for providing a host cell for use in the present invention. The host cell may be a prokaryotic cell, such as a eubacterial or archaebacterial cell; or a eukaryotic cell, such as a plant cell, an animal cell, an insect cell, a fungal cell or a yeast cell. Practically all eukaryotic cells produce GGPP for their biosynthesis, but in some embod- iments a eukaryotic cell produces an increased amount of GGPP, which may increase the production of miltiradiene, compared with a similar eukaryotic that does not produce increased amounts of GGPP. Methods for increasing the GGPP production in a eukary- otic cell has also been described in the prior art. The host cell may be a unicellular organism, or it may be comprised within a multicellular organism, e.g. a plant. Examples of suitable plants or plant cells for use as host cells according to the invention includes corn (Zea mays), canola (Brassica napus, Brassica rapa ssp.), alfalfa (Medicago sativa), rice (Oryza sativa), rye (Secale cerale), sorghum (Sorghum bicolor, Sorghum vulgare), sunflower (Helianthus annuas), wheat (Tritium aestivum and other species), triticale, rye (Secale) soybean (Glycine max), tobacco (Ni- cotiana tabacum), potato (Solanum tuberosum), peanuts (Arachis hypogaea), cotton (Gossypium hirsutum), sweet potato (Impomoea batatus), cassava (Manihot esculenta), coffee (Coffea spp.), coconut (Cocos nucifera), pineapple (Anana comosus), citrus (Cit- rus spp.) cocoa (Theobroma cacao), tea (Camellia senensis), banana (Musa spp.), ava- cado (Persea americana), fig (Ficus casica), guava (Psidium guajava), mango (Mangifer indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occi- dentale), macadamia (Macadamia intergrifolia), almond (Primus amygdalus), apple (Ma- lus spp.), pear (Pyrus spp.), plum and cherry tree (Prunus spp.), ribes (currant etc.), Vitis, Jerusalem artichoke (Helianthemum spp.), non-cereal grasses (Grass family), sugar and fodder beets (Beta vulgaris), chicory, oats, barley, vegetables, or ornamentals, crop plants (for example, cereals and pulses, maize, wheat, potatoes, tapioca, rice, sorghum, millet, cassava, barley, pea, sugar beets, sugar cane, soybean, oilseed rape, sunflower and other root, tuber or seed crops. Other important plants may be fruit trees, crop trees, forest trees or plants grown for their use as spices or pharmaceutical products (Mentha spp., clove, Artemesia spp., Thymus spp., Lavendula spp., Allium spp., Hypericum, Catharanthus spp., Vinca spp., Papaver spp., Digitalis spp., Rawolfia spp., Vanilla spp., Petrusilium spp., Eucalyptus, tea tree, Picea spp., Pinus spp., Abies spp., Juniperus spp. Horticultural plants which can be used with the present invention may include lettuce, endive, and vegetable brassicas including cabbage, broccoli, and cauliflower, carrots, and carnations and geraniums. The plant can also be tobacco, cucurbits, carrot, strawberry, sunflower, tomato, pepper, or chrysanthemum. Further examples of plants include grain plants for example oil-seed plants or legumi- nous plants. Seeds of interest include grain seeds, such as corn, wheat, barley, sorghum, rye, etc. Oil-seed plants include cotton soybean, safflower, sunflower, Brassica, maize, alfalfa, palm, coconut, etc. Leguminous plants include beans and peas. Beans include guar, locust bean, fenugreek, soybean, garden beans, cowpea, mung bean, lima bean, fava bean, lentils, chickpea. Particular preferred plant species include Physcomitrella sp., such as P. patens; Ara- bidopsis sp., such as A. thaliana; Nicotiana sp., such as N. benthamiana; Chlamydomo- nas sp., such as C. reinhardtii; and Nannochloropsis sp., such as N. oceanica. Examples of suitable eukaryotic cell for use according to the invention include fungal cells such as Agaricus, Aspergillus, Candida, Eremothecium, Fusarium/Gibberella, Kluy- veromyces, Laetiporus, Lentinus, Phaffia, Phanerochaete, Pichia, Physcomitrella, Rhodoturula, Saccharomyces, Schizosaccharomyces, Sphaceloma, Xanthophyllomy- ces or Yarrowia. Exemplary species from such genera include Lentinus tigrinus, Laeti- porus sulphureus, Phanerochaete chrysosporium, Pichia pastoris, Cyberlindnera jadinii, Physcomitrella patens, Rhodoturula glutinis, Rhodoturula mucilaginosa, Phaffia rhodozyma, Xanthophyllomyces dendrorhous, Fusarium fujikuroi/Gibberella fujikuroi, Candida utilis, Candida glabrata, Candida albicans, and Yarrowia lipolytica. In some embodiments, a host cell can be an Ascomycete such as Gibberella fujikuroi, Kluyveromyces lactis, Schizosaccharomyces pombe, Aspergillus niger, Yarrowia lipo- lytica, Ashbya gossypii, or S. cerevisiae. In some embodiments, the host cell can be an algae cell such as Blakeslea trispora, Dunaliella salina, Haematococcus pluvialis, Chlorella sp., Undaria pinnatifida, Sargas- sum, Laminaria japonica, Scenedesmus almeriensis. In some embodiments, a host cell can be a prokaryote such as Bacillus cells, for example Bacillus subtilis; Escherichia cells, for example, Escherichia coli cells; Lactobacillus cells; Lactococcus cells; Streptomyces cells, Streptococcus cells, Cornebacterium cells; Acetobacter cells; Acinetobacter cells; or Pseudomonas cells. In some embodiments, the host cell can be a cyanobacterial cell such as Synechocystis sp. or Synechococcus sp. In one embodiment, a host cell that is suitable for growth in a fermenter is selected. Growing the recombinant host cell according to the invention is a convenient way of growing the host cell for production of the oxygenated diterpenoid compounds of the invention. In another embodiment, the host cell is a phototropic cell and the cell is cultivated in a green house or photobioreactor. The genes and enzymes The recombinant host cell of the present invention is capable of producing miltiradiene and/or dehydroabietadiene. Miltiradiene may be spontaneously converted into dehydro- abietadiene or it may be converted by an enzyme that facilitates the conversion of milti- radiene to dehydroabietadiene. As described herein above, the synthesis of miltiradiene usually begins with the for- mation of GGPP by condensation of one dimethylallyl pyroophosphate (DMAP) mole- cule and three isopentenyl pyrophosphate (IPP) molecules by a geranylgeranyl diphos- phate synthase. Recombinant host cells and heterologous nucleic acids that encode enzymes that cata- lyze the synthesis of GGPP in recombinant host cells are generally known in the art, see e.g. WO 2015/113570. In addition, many host organisms are capable of producing GGPP intrinsically and heterologous nucleic acids may thus not always be necessary for production of GGPP. In some embodiments, the recombinant host cell comprises a heterologous nucleic acid encoding a geranylgeranyl diphosphate synthase, such as the geranylgeranyl di- phosphate synthase SpGGPPs7 as set forth in SEQ ID NO: 73 or SEQ ID NO: 81, or a functional homologue thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the ma- ture polypeptide thereof. Subsequently, GGPP may be converted into miltiradiene by the action of one or more diterpene synthases, copalyl diphosphate synthases and/or miltiradiene synthases. In some embodiments, the recombinant host cell comprises one or more heterologous nucleic acids encoding one or more diterpene synthases, such as the diterpene syn- thases CfTPS1 (SEQ ID NO: 67) and CfTPS3 (SEQ ID NO: 68), or CftTPS1 (SEQ ID NO: 77) and CftTPS3 (SEQ ID NO: 78), or respective functional homologues thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptides thereof. In some embodiments, the recombinant host cell comprises one or more heterologous nucleic acids encoding one or more diterpene synthases, such as the diterpene syn- thases TwTPS9 (SEQ ID NO: 69) and TwTPS27 (SEQ ID NO: 70), or respective func- tional homologues thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptides thereof. In some embodiments, the recombinant host cell comprises a combination of one or more copalyl diphosphate synthases and one or more miltiradiene synthases, such as a combination of the copalyl diphosphate synthases SmCPS (SEQ ID NO: 71) and the miltiradiene synthase SmKSL (SEQ ID NO: 72), or respective functional homologues thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptides thereof. In an even further aspect, the invention relates to polypeptides having cytochrome P450 enzyme activity and comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, preferably at least 96% sequence identity, preferably at least 97% sequence identity, preferably at least 98% sequence identity or even 100% sequence identity to one of the sequences SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7 or the mature polypeptide thereof. In a further aspect, the invention relates to a polypeptide having cytochrome B5 activity and comprising an amino acid sequence having at least 80 % sequence identity, prefer- ably at least 85% sequence identity, preferably at least 90% sequence identity, prefera- bly at least 95% sequence identity, preferably at least 96% sequence identity, preferably at least 97% sequence identity, preferably at least 98% sequence identity or even 100% sequence identity to SEQ ID NO: 8, or the mature polypeptide thereof. The invention also relates to polynucleotide sequences or genes encoding polypeptides having cytochrome P450 enzyme activity and comprising an amino acid sequence hav- ing at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, preferably at least 96% sequence identity, preferably at least 97% sequence identity, preferably at least 98% sequence identity or even 100% sequence identity to one of the sequences SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7 or the mature polypeptide thereof, or encoding a polypeptide having cytochrome B5 activity and comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, preferably at least 96% sequence identity, preferably at least 97% sequence identity, preferably at least 98% sequence identity or even 100% sequence identity to SEQ ID NO: 8, or the mature polypeptide thereof. In preferred embodiments, one or more of the first, second, third, fourth, fifth and sixth enzymes having cytochrome P450 activity comprises or consists of an amino acid se- quence according to any one of SEQ ID NO: 1 (TwCYP82D274v1), SEQ ID NO: 2 (TwCYP82D274v2), SEQ ID NO: 74 (TwCYP82D274v3), SEQ ID NO: 75 (TwCYP82D274v4), SEQ ID NO: 3 (TwCYP71BE85), SEQ ID NO: 4 (TwCYP71BE86), SEQ ID NO: 5 (TwCYP82D213v1) and SEQ ID NO: 76 (TwCYP82D213v2), and re- spective functional homologs thereof having at least 80% sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, preferably at least 96% sequence identity, preferably at least 97% sequence identity, preferably at least 98% sequence identity, preferably at least 98% sequence identity thereto, or the mature polypeptide thereof. In some embodiments, the first heterologous nucleic acid encoding a first enzyme hav- ing cytochrome P450 activity encodes TwCYP82D274 as set forth in SEQ ID NO: 1 (TwCYP82D274v1, SEQ ID NO: 2 (TwCYP82D274v2), SEQ ID NO: 74 (TwCYP82D274v3), SEQ ID NO: 75 (TwCYP82D274v4), or a functional homologue thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto. In some embodiments, the second heterologous nucleic acid encoding a second en- zyme having cytochrome P450 activity encodes the cytochrome P450 enzyme TwCYP71BE86 as set forth in SEQ ID NO: 4, or a functional homologue thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto. In some embodiments, the third heterologous nucleic acid encoding a third enzyme having cytochrome P450 activity encodes the cytochrome P450 enzyme TwCYP71BE85 as set forth in SEQ ID NO: 3, or a functional homologue thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto. In some embodiments, the fourth heterologous nucleic acid encoding a fourth enzyme having cytochrome P450 activity encodes the cytochrome P450 enzyme TwCYP82D213v1 as set forth in SEQ ID NO: 5 or TwCYP82D213v2 as set forth in SEQ ID NO: 76 (TwCYP82D213v2), or a functional homologue thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto. In some embodiments, the fifth heterologous nucleic acid encoding a fifth enzyme hav- ing cytochrome P450 activity encodes the cytochrome P450 enzyme TwCYP82D217 as set forth in SEQ ID NO: 6, or a functional homologue thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% se- quence identity thereto. In some embodiments, the sixth heterologous nucleic acid encoding a sixth enzyme having cytochrome P450 activity encodes the cytochrome P450 enzyme TwCYP82D275 as set forth in SEQ ID NO: 7, or a functional homologue thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto. In some embodiments, the seventh heterologous nucleic acid encoding an enzyme having cytochrome B5 activity encodes the cytochrome B5 enzyme TwB5#1 as set forth in SEQ ID NO: 8, or a functional homologue thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto. In some embodiments is provided a recombinant host cell i. wherein the host cell is capable of producing miltiradiene and/or dehydroabietadi- ene; and ii. comprises a heterologous nucleic acid encoding TwCYP82D274 of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 74 or SEQ ID NO: 75, or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof, wherein said cell is capable of producing 14-hydroxydehydroabietadiene. In some embodiments is provided a recombinant host cell i. wherein the host cell is capable of producting miltiradiene and/or dehydro- abietadiene; and ii. comprises a heterologous nucleic acid encoding TwCYP82D274 of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 74 or SEQ ID NO: 75, or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; and iii. comprises a heterologous nucleic acid encoding TwCYP71BE86 of SEQ ID NO: 4 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof, wherein said cell is capable of producing 14-hydroxydehydroabietadiene, 3,14-dihy- droxydehydroabietadiene, 3,14-dihydroxyabeodiene and/or 14-hydroxy-18-aldo-abeo- diene. In some embodiments is provided a recombinant host cell i. wherein the host cell is capable of producting miltiradiene and/or dehydro- abietadiene; and ii. comprises a heterologous nucleic acid encoding TwCYP82D274 SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 74 or SEQ ID NO: 75, or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; iii. comprises a heterologous nucleic acid encoding TwCYP71BE86 of SEQ ID NO: 4 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; and iv. comprises a heterologous nucleic acid encoding TwCYP71BE85 of SEQ ID NO: 3 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof, wherein said cell is capable of producing 14-hydroxydehydroabietadiene, 3,14-dihy- droxydehydroabietadiene, 3,14-dihydroxyabeodiene, 14-hydroxy-18-aldo-abeodiene and/or triptophenolide. In some embodiments is provided a recombinant host cell i. wherein the host cell is capable of producting miltiradiene and/or dehydro- abietadiene; and ii. comprises a heterologous nucleic acid encoding TwCYP82D274 of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 74 or SEQ ID NO: 75, or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; iii. comprises a heterologous nucleic acid encoding TwCYP71BE86 of SEQ ID NO: 4 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; iv. comprises a heterologous nucleic acid encoding TwCYP71BE85 of SEQ ID NO: 3 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; and v. comprises a heterologous nucleic acid encoding TwCYP82D213 of SEQ ID NO: 5 or SEQ ID NO: 76, or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof, wherein said cell is capable of producing 14-hydroxydehydroabietadiene, 3,14-dihy- droxydehydroabietadiene, 3,14-dihydroxyabeodiene, 14-hydroxy-18-aldo-abeodiene, triptophenolide and/or triptonide. In some embodiments is provided a recombinant host cell i. wherein the host cell is capable of producting miltiradiene and/or dehydro- abietadiene; and ii. comprises a heterologous nucleic acid encoding TwCYP82D274 of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 74 or SEQ ID NO: 75, or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; iii. comprises a heterologous nucleic acid encoding TwCYP71BE86 of SEQ ID NO: 4 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; iv. comprises a heterologous nucleic acid encoding TwCYP71BE85 of SEQ ID NO: 3 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; v. comprises a heterologous nucleic acid encoding TwCYP82D213 of SEQ ID NO: 5 or SEQ ID NO: 76, or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; and vi. comprises a heterologous nucleic acid encoding TwB5#1 of SEQ ID NO: 8 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% se- quence identity thereto, or the mature polypeptide thereof, wherein said cell is capable of producing triptonide with a titer that is at least 2-fold, such as at least 3-fold, such as at least 4-fold, such as at least 5-fold higher than an identical yeast cell, except wherein said yeast said does not express said TwB5#1 or said functional homolog thereof. In some embodiments is provided a recombinant host cell i. wherein the host cell is capable of producting miltiradiene and/or dehydroabietadi- ene; and ii. comprises a heterologous nucleic acid encoding TwCYP82D274 of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 74 or SEQ ID NO: 75,or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; iii. comprises a heterologous nucleic acid encoding TwCYP71BE86 of SEQ ID NO: 4 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; iv. comprises a heterologous nucleic acid encoding TwCYP71BE85 of SEQ ID NO: 3 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; v. comprises a heterologous nucleic acid encoding TwCYP82D213 of SEQ ID NO: 5 or SEQ ID NO: 76, or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% sequence identity thereto, or the mature polypeptide thereof; and vi. comprises a heterologous nucleic acid encoding TwB5#1 of SEQ ID NO: 8 or a functional homolog thereof having at least 80%, such as at least 81%, such as at least 82%, such as at least 83%, such as at least 84%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 91%, such as at least 92%, such as at least 93%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% se- quence identity thereto, or the mature polypeptide thereof, wherein said cell is capable of growing in a fermentation medium and where said fer- mentation medium after 7 days of fermentation comprises: - at least 3 ppm triptonide and/or - at least 1 ppm triptophenolide. Aforementioned recombinant host cells may be capable of producing miltiradiene and/or dehydroabietadiene for several different reasons. For example, the host cells may endogenously be capable of producing miltiradiene. Alternatively, the recombinant host cell may comprise one or more heterologous nucleic acid sequences encoding one or more enzymes involved in the production of miltiradiene, such as the diterpene biosynthetic enzymes SPGGPPS7 of SEQ ID NO: 73 or SEQ ID NO: 81, CfTPS1 of SEQ ID NO: 67, CftTPS1 of SEQ ID NO: 77, CfTPS3 of SEQ ID NO: 68, CftTPS3 of SEQ ID NO: 78 and/or TwCPR1 of SEQ ID NO: 9, or respective functional homologs thereof having at least 80% sequence identity, preferably at least 85% sequence iden- tity, preferably at least 90% sequence identity, preferably at least 95% sequence iden- tity, preferably at least 96% sequence identity, preferably at least 97% sequence iden- tity, preferably at least 98% sequence identity, preferably at least 98% sequence iden- tity thereto, or the mature polypeptides thereof. Functional homologues of the first (e.g. TwCYP82D274), second (e.g. TwCYP71BE86), third (e.g. TwCYP71BE85), fourth (e.g. TwCYP82D213), fifth (e.g.TwCYP82D217) and sixth (e.g.TwCYP82D275) enzymes having cytochrome P450 activity and the enzyme having cytochrome B5 activity (e.g.TwB5#1) may be verified by expressing the relevant protein in a yeast cell and assessing whether they are able to produce specific compounds as described herein below. A yeast cell expressing a functional homolog of TwCYP82D274 (SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 74 or SEQ ID NO: 75) and further expressing i. the diterpene biosynthetic enzymes SPGGPPS7v2 (SEQ ID NO: 81), CftTPS1 (SEQ ID NO: 77), CftTPS3 (SEQ ID NO: 78) and TwCPR1 (SEQ ID NO: 9), is preferably capable of producing 14-hydroxydehydroabietadiene. A yeast cell expressing a functional homolog of TwCYP71BE86 (SEQ ID NO: 4) and further expressing i. the diterpene biosynthetic enzymes SPGGPPS7v2 (SEQ ID NO: 81), CftTPS1 (SEQ ID NO: 77), CftTPS3 (SEQ ID NO: 78) and TwCPR1 (SEQ ID NO: 9); and ii. TwCYP82D274 (SEQ ID NO: 1 or SEQ ID NO: 2), is preferably capable of producing 14-hydroxydehydroabietadiene, 3,14-dihydroxydehy- droabietadiene, 3,14-dihydroxyabeodiene and 14-hydroxy-18-aldo-abeodiene. A yeast cell expressing a functional homolog of TwCYP71BE85 (SEQ ID NO: 3) and further expressing i. the diterpene biosynthetic enzymes SPGGPPS7v2 (SEQ ID NO: 81), CftTPS1 (SEQ ID NO: 77), CftTPS3 (SEQ ID NO: 78) and TwCPR1 (SEQ ID NO: 9); ii. TwCYP82D274 (SEQ ID NO: 1 or SEQ ID NO: 2); and iii. TwCYP71BE86 (SEQ ID NO: 4), is preferably capable of producing 14-hydroxydehydroabietadiene, 3,14-dihydroxydehy- droabietadiene, 3,14-dihydroxyabeodiene, 14-hydroxy-18-aldo-abeodiene and tripto- phenolide. A yeast cell expressing a functional homolog of TwCYP82D213 (SEQ ID NO: 5 or SEQ ID NO: 76) and further expressing i. the diterpene biosynthetic enzymes SPGGPPS7v2 (SEQ ID NO: 81), CftTPS1 (SEQ ID NO: 77), CftTPS3 (SEQ ID NO: 78) and TwCPR1 (SEQ ID NO: 9); ii. TwCYP82D274 (SEQ ID NO: 1 or SEQ ID NO: 2); iii. TwCYP71BE86 (SEQ ID NO: 4); and iv. TwCYP71BE85 (SEQ ID NO: 3), is preferably capable of producing 14-hydroxydehydroabietadiene, 3,14-dihydroxydehy- droabietadiene, 3,14-dihydroxyabeodiene, 14-hydroxy-18-aldo-abeodiene, triptophe- nolide and triptonide. A yeast cell expressing a functional homolog of TwB5#1 (SEQ ID NO: 8) and further expressing i. the diterpene biosynthetic enzymes SPGGPPS7v2 (SEQ ID NO: 81), CftTPS1 (SEQ ID NO: 77), CftTPS3 (SEQ ID NO: 78) and TwCPR1 (SEQ ID NO: 9); ii. TwCYP82D274 (SEQ ID NO: 1 or SEQ ID NO: 2); iii. TwCYP71BE86 (SEQ ID NO: 4); iv. TwCYP71BE85 (SEQ ID NO: 3); and v. TwCYP82D213 (SEQ ID NO: 5 or SEQ ID NO: 76) is preferably capable of producing triptonide with a titer that is at least 2-fold, such as at least 3-fold, such as at least 4-fold, such as at least 5-fold higher than an identical yeast cell, except wherein said yeast said does not express said functional homolog of TwB5#1. In preferred embodiments, the enzyme having cytochrome B5 activity comprises or consists of an amino acid sequence according to SEQ ID NO: 8 (TwB5#1), or a func- tional homolog thereof having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, preferably at least 96% sequence identity, preferably at least 97% sequence identity, preferably at least 98% sequence identity, preferably at least 98% sequence identity thereto, or the mature polypeptide thereof. The polynucleotide of the invention may be provided by cloning from organisms that nat- urally produce the polypeptides such as the plant T. wilfordii or closely related plants, or it may be provided by chemical synthesis of the polynucleotide sequence based on tech- niques known in the art. The polynucleotide may have a sequence that is identical to a sequence found in nature, or it may have a sequence that is not found in nature, e.g. the sequence may be codon optimized for the particular selected host cell. The polypeptides of the invention may be provided from organisms that naturally produce the polypeptides such as the plant T. wilfordii or related organisms; or they may be pro- vided by inserting and expressing polynucleotides encoding polypeptides into a suitable host cell and recovering the polypeptides from culture broth comprising the host cell transformed with the respective genes. It is preferred to provide the polypeptides of the invention from a suitable selected recombinant host cell. In order to transform and express a gene in a suitable host cell, the gene is usually operably connected with suitable regulatory elements and inserted into an expression vector suitable for the particular selected host cell. Selecting suitable regulatory ele- ments, constructing a suitable expression vector and transforming the selected host cell is within the skills of the average practitioner and the invention is not limited by any par- ticular selection of such elements. The generated host cells comprising the genes of the invention are suitable grown in a container, e.g. a fermenter or shake flasks; under conditions where the genes are ex- pressed and the oxygenated diterpenoid compounds are formed. When growth ceases or a sufficiently high amount of the oxygenated diterpenoid compounds are accumulated in the culture broth, the oxygenated diterpenoid compounds may be further modified and recovered from the culture broth. Sequence identity is understood as a measurement of the similarity between two amino acid or nucleotide sequences. Sequence identity is calculated by first aligning the two sequences, counting the number of positions where the two sequences contain the same amino acid residue or nucleotide and calculating the percent identity as the number of positions with identical amino acid residue or nucleotide with the whole length of the alignment. Several algorithms have been developed and are available for the skilled person. In this specification and claim, the sequence identity for amino acid sequences are cal- culated using the NCBI BLAST+ pairwise alignment algorithm, using default parameters (BLOSUM 62 matrix, Gap open penalty 11; gap extend penalty 1, Exp. Thr 10), and the sequence identity for nucleotide sequences are calculated using the NCBI BLAST+ pair- wise alignment algorithm, using default parameters (Match/mismatch scores 1, -3; gap open penalty 5; gap extend penalty 2; exp. Thr 10). The NCBI BLAST+ programs are further described in: Madeira F el at (2019) NAR 47: W636-W641. EXAMPLES Materials and Methods Genetic engineering of Nicotiana benthamiana Tripterygium wilfordii CYP genes were cloned from plant material and co-expressed in Nicotiana benthamiana with the diterpene biosynthesis genes CfDXS (SEQ ID NO: 79) or SctHMGR (SEQ ID NO: 80), CfGGPPS or SpGGPPS7 (SEQ ID NO: 81), CfTPS1 (SEQ ID NO: 67) or CftTPS1 (SEQ ID NO: 77), and CfTPS3 (SEQ ID NO: 68) or CftTPS3 (SEQ ID NO: 78) using constructs and methods previously described in (1-4). Also co- expressed, were the suppressor of gene silencing, p19. Briefly, binary vectors each con- taining individual diterpene biosynthesis genes or Tripterygium wilfordii CYPs (TwCYPs) were transformed into agrobacteria. Liquid cultures of agrobacteria each containing spe- cific plasmids were mixed for co-expression of specific combinations of TwCYPs. Genetic engineering of Saccharomyces cerevisiae and growth conditions for engineered S. cerevisiae Media YPD media: 20 g/L Bacto™ Peptone, 10 g/L Bacto™ Yeast extract, 20 g/L glucose. Synthetic complete (SC) meda without uracil: 1.92 g/L Yeast Synthetic Drop-out Media Supplements without uracil (Sigma-Aldrich Co. LLC. Catalog number Y1501), 6.7 g/L Yeast Nitrogen Base Without Amino Acids (Sigma-Aldrich Co. LLC. Catalog number Y0626), 20 g/L glucose. Feed-In-Time (FIT) was based on EnPump200 (Enpresso GmbH), and made according to protocol enclosed with the product. Agar plates: SC me- dia including agar (15 g/L). Uracil auxotrophy in parent strains was introduced by selecting for lack of URA3 function on agar plates of SC medium without uracil containing also 5-Fluoroorotic Acid (5-FOA, 0.74 g/L) and uracil (30 mg/L). Yeast transformants were isolated on SC without uracil agar plates. Feed-batch fermentation of engineering S. cerevisiae strains for isolation of miltiradiene derived diterpenoids All engineered S. cerevisiae strains were cultivated in 96-deepwell plates using a Feed- In-Time (FIT; m2p-labs) approach similar to previously described (insert ref Forman et al. 2018). For isolation and purification of key intermediates in the triptonide pathway selected engineered S. cerevisiae strain were cultivated in feed batch fermentor using a 2 L Biostat® A bioreactor (Sartorius AG). Fed batch fermentation was initiated by addi- tion of a 100 mL starter culture to the reactor tank (with impellers), which in turn was prepared earlier by autoclavation while containing 200 mL Batch glucose and 300 mL Batch salt mix. Also 5 mL vitamin mix, 5 mL micro elements and 0.5 mL trace elements, were added. Cultivation in the bioreactor was started under the following conditions (monitored and automatically controlled): pH = 5, temp. = 30 °C, dissolved oxygen (DO) = 20 %. While pH was controlled by feeding of ammonium hydroxide (32 %) and sulfuric acid (10 %), dissolved oxygen was controlled by air supply combined with stirring. Also foam levels were adjusted by addition of anti-foam emulsion (35119, Serva Electropho- resis GmbH). After 18 hours of initial cultivation in the bioreactor, feeding with Feeding solution at a rate of 1.3 % was started. The fermentation process continued for 7 days with daily sampling of the culture. Extraction of engineered S. cerevisiae for LC-MS analysis Genetically engineered S. cerevisiae strain was transferred into 0.5mL media in a 96- well plate and grown for 3 days at 30 ºC with orbital shaking at 350 rpm. For extraction 0.1mL of S. cerevisiae culture was transferred to 1.5mL glass vials.0.4mL MeOH uHPLC grade was added. S. cerevisiae extract was filtered by using a 0:22 μm 96-well filter plate (Merck Millipore, Darmstadt, Germany) and at stored at 4 ^C prior to LC-MS analysis. Extraction of diterpenoid metabolites for LC-MS analysis Samples of yeast cultures for LCMS analysis were prepared in 1.5 mL glass vials by mixing yeast cultures and methanol spiked with 5 ppm andrographolide (internal stand- ard; FA17902, CarboSynth) in a ratio of 1:19 (v/v) for daily bioreactor samples and 1:4 (v/v) for 96-deepwell cultures. Mixing proceeded for 30 min with shaking at room temp. For tobacco samples, 2 leaf discs (ø=3cm) placed in 1.5 ml glass vials, were extracted with 1 mL of the methanol extraction solution, for 1 h with shaking at room temperature. Before LCMS analysis samples were passed through a 0.22 µM 96-well plate filter (Merck Millipore, Darmstadt, Germany) and stored at 5°C. LC-MS analysis: Methanol (MeOH) extracts were analysed using an Ultimate 3000 UHPLC+ Focused system (Dionex Corporation, Sunnyvale, CA) coupled to a Bruker Compact ESI-QTOF- MS (Bruker) system. Samples were separated on a Kinetex XB-C18 column (100 x 2.1 mm ID, 1:7 μm particle size, 100 °A pore size; Phenomenex Inc., Torrance, CA) main- tained at 40 _C with a flow rate of 0.3 mL min-1 and mobile phase consisting of 0.05 % (v/v) formic acid in water (solvent A) and 0.05 % (v/v) formic acid in acetonitrile (solvent B). Two LC protocols were used: LC method 1: 0-0.5 min, 10 % B; 0.5-21 min, linear increase from 10 to 80 % B; 21-31 min, to 90 % B; 31-34 min, to 100 % B; 34-39 min 100% B; 39-40 min linear decrease from 100 to 10 % B. LC method 2: 0-0.5 min, 20 % B; 0.5-11 min, linear increase from 20 to 80 % B; 11-20 min, to 90 % B; 20-22 min, to 100 % B; 22-27 min 100% B; 27-28 min linear decrease from 100 to 20 % B. LC method 3: 0-0.5 min, 20 % B; 0.5-9 min, linear increase from 20 to 100 % B; 9-11 min, 100% B; 11-11.5 min, linear decrease from 100 to 20 % B; 11.5-15min, 20% B. Extraction of diterpenoid metabolites for GC-MS analysis Samples of yeast cultures for GCMS analysis were prepared in 1.5 mL glass vials by mixing yeast culture and pure methanol at a ratio of 1:4 (v/v). After brief mixing, apolar constituents, were liquid-liquid extracted into hexane, spiked with 10 ppm 1-eicocene, by mixing at a ratio of 1:1 (v/v) and shaking for 1 h. For tobacco samples, 2 leaf discs (ø=3cm) placed in 1.5 ml glass vials, were extracted with 1mL of the same hexane solu- tion, via 1 h of shaking. Prior to GCMS analysis hexane layers were transferred to new vials. Gas chromatography Mass spectrometry (GC-MS) analysis GC-MS analysis was carried out on a Shimadzu GCMS-QP2010 Ultra (Shimadzu Corp.) with an Agilent HP-5MS column (Agilent Technologies) 20 m x 0.18 mm i.d., 0.18 µm film thickness). Hydrogen was used as a carrier gas at a constant linear velocity of 50 cm s-1, and the injection volume was 1 µL at 250°C (splitless mode). The oven program was 80°C for 2 min, ramp at rate 20 °C/min to 180°C, ramp at rate 10 °C/min to 300 °C, ramp at rate 20 °C/min to 310°C, hold for 3 min. Data was stored in .CDF format and processed in MZmine2. Relative quantification of miltiradiene derived diterpenoids Relative compound quantities in yeast cultures were based on normalized peak areas of characteristic ions (data obtained using targeted feature detection in the MZmine2 soft- ware). The signal for the following ions were quantified: 1: miltiradiene m/z 91.1, 2: 14- hydroxyabietadiene m/z 189.1, 3: F15P1 m/z 303.2318, 4: F20P2 m/z 283.2059, 5: F15P2 m/z 299.2002, 6: triptophenolide m/z 313.1794, 8: triptonide m/z 359.1481. For LCMS and GCMS data a mass deviation of 5ppm and 100ppm, respectively, was toler- ated. The peak area of the base peak ion (m/z 315.1947) for the internal standard andro- grapholide was used for normalization. Absolute quantifications Absolute quantifications of triptophenolide (FT65732, CarboSynth) and triptonide (FT65197, CarboSynth) were done by co-analysis of authentic standards prepared in methanol and a final concentration of 5 ppm internal standard (andrographelide). Quantification was based on normalized peak area and calculated from the slopes of linear extrapolations of the standards response curve (triptophenolide 0.05, 0.5, 1, 2 ppm; triptonide 0.5, 1, 2, 10, 20 ppm). Isolation and purification of miltiradiene derived compounds from engineered S. cere- visiae strain for NMR analysis Compounds in this invention were isolated from bioreactor cultures yeast strains NVJ8.15, and NVJ3.10, and structurally elucidated by NMR. The combined ethyl acetate extracts of broth and methanol-lysed cells (cells:methanol = 1:4, v/v) were initially dried in presence of Celite S® (06858, Sigma-Aldrich) via rotary evaporation. Compounds were subsequently isolated by successive fractionations using a puriFlash® 5.250 (In- terchim, Montluçon, France) instrument with detection by UV absorbance and Evapora- tive Light-Scattering Detection (ELSD). This was equipped with either of columns (C1) PF-15SIHP-F0025 (OV002A, Interchim) and (C2) US5C18HQ-100/300 (SSP750, Inter- chim) for normal phase- and reverse phase separation, respectively. An initial pre-fractionation of the dry mix of Celite S®/crude extract was achieved using column (ref.9) with loading from a manually packed dry-loading column. Separation was obtained using mobile phases hexane (A) and ethyl acetate (B), a constant flow rate of 15 mL/min, followed by a final washing step with 100% methanol. Compounds of interest were detected by UV and ELSD and collected. Collected fractions were continuously evaluated by LCMS using LC-MS method 3 and TLC analysis prior to further fractionation or NMR studies. Additional purification of compounds of interest from fraction with multi- ple compounds was done by an additional normal phase fraction using C1 or a reverse phase column fractionation using C2. For reverse phase purification with C2 samples were evaporated using rotor evaporation and resuspended in 2mL methanol. Sample was injected directly onto the pre condi- tioned column C2. Mobile phases for C2 consisted of solvent C: deionized water and solvent D acetonitrile each acidified with 0.05% (v/v) formic acid. A constant flow rate of 32 mL/min, was used, with a linear solvent gradient with increasing concentration of sol- vent D. Compounds of interest were detected by ELSD and UV and collected. Additional reverse phase purification was done by multiple injections of 100uL ontp a semi-prep Phenomenex Luna 5µm C18(2) 100Å 250x10mm (fully porous)(Phenomenex, Inc., Torrance, CA, USA) column on a Shimadzu HPLC (SPD-M20A diode array detec- tor, FRC-10A fraction collector, DGU-20A5 degasser, LC-20AT pump, CBM-20A System controller, CTO-10AS VP column oven, SIL-10AP autosampler). Mobile phase was a linear gradient between C and D with an increasing amount of D going from 50-100%. Compounds of interest was detected by UV absorbance at 210nm and collected. Mass spectra Mass spectra were acquired in positive ion mode over a scan range of m/z 50-1200 with the following ESI and MS settings: capillary voltage, 4000 V; end plate offset, 500 V; dry gas temperature, 220 ^C; dry gas flow of 8 L min1; nebulizer pressure, 2 bar; in source CID energy, 0 eV; hexapole RF, 50 Vpp; quadrupole ion energy, 4 eV; collision cell en- ergy, 7 eV. Raw chromatogram data was calibrated using an internal sodium formate standard and subsequently exported as mzML format using DataAnalysis 4.3 (Build 110.102.1532) (64-bit), Bruker. MZmine ver 2.53 was used for visualizing the LC-MS chromatograms. Media recipes for bioreactor starting media and feed media Batch glucose: Glucose monohydrate 55 g/L Batch salt mix: Ammonium sulfate 25 g/L Potassium phosphate monobasic 5 g/L Magnesium sulfate heptahydrate 1.7 g/L Feed glucose: Glucose monohydrate 880 g/L Feed salt mix: Potassium phosphate monobasic 21.6 g/L Magnesium sulfate heptahydrate 24.24 g/L Potassium sulfate 8.4 g/L Sodium sulfate 0.672 g/L Preparation notes: Batch- and feed salt mixes as well as batch and feed glucose were prepared in separate BlueCap bottles by dissolving components in Milli-Q water and sterilizing by autoclava- tion. Feeding solution: A feeding solution was made by mixing 500 mL of feed glucose with 500 mL of feed salt mix, 10 mL of vitamin mix, 10 mL of micro elements solution and 1 mL of trace elements solution. Example 1: Expression in Nicotiana benthamiana Leaf material of N. benthamiana co-expressing specific combinations of genes of interest (GOI) was harvested 7 days after agrobacterial infiltration.1mL methanol (MeOH) was added to 2 leaf disks (Ø = 2cm). Extraction was done at room temperature at 200rpm orbital shaking. 200 uL of extract was filtered by using a 0:22 μm 96-well filter plate (Merck Millipore, Darmstadt, Germany) and at stored at 4 ^C prior to LC-MS analysis. Figure 1 shows the obtained LCMS profiles. The results show that the N. benthamiana cells transformed with CYP82D274V1 encoding the enzyme having SEQ ID NO: 1, leads to production of 14-OH-dehydroabietadiene; when the cells are further transformed with CYP71BE85 and CYP71BE86 encoding the enzymes having the amino acid sequences SEQ ID NO: 3 and SEQ ID NO: 4, respectively, triptophenolide is formed; and when the cells are further transformed with CYP82D213 encoding the enzyme having the amino acid sequence of SEQ ID NO: 5 triptonide is formed. Further, it can be seen that the enzyme having the sequence of SEQ ID NO:6 and encoded by the gene CYP82D217 increases the production of triptophenolide and triptonide. Example 2: Construction of S. cerevisiae strains. Strain construction Parent yeast strain was S. cerevisiae S288C (NCYC 3608; National Collection of Yeast Cultures, Norwich, UK). Genotypes and source of strains are listed in table 3. Constructed yeast strains were made using the lithium acetate transformation method (8). Parent strains without functional URA3 were made competent by the following pro- cedure: Inoculation from a glycerol stock into 5 ml YPD medium and growing at 30⁰C O/N. Then, transfer of 3 mL of O/N culture to 50 mL YPD medium and continued growing for 4-5 hours followed by centrifugation at 4000 RPM for 10 minutes then discarding the supernatant. Cells were then ready for transformation after 2 washes in sterile water (1st in 25 mL, 2nd in 1 mL) and resuspension in 0.4 mL of sterile water. Transformation of competent yeast cells was done by the following procedure: Mixes of designated NotI digested plasmids (2 µL of each) were each added 10 µL competent yeast cells and mixed with 60 µL PEG 3350 (50% w/v), 9 µL LiAc (1 M) and 12.5 µL preboiled salmon sperm DNA (10 mg/ml). The resulting mixes were next incubated at 42⁰C for 40 minutes before cells were collected by centrifugation (3000 RPM for 5 minutes) and removal of supernatant. Cells were then resuspended in 100 µL sterile water and spread on SC without uracil agar plates. Isolated transformants appeared as single colonies after 2 days of incubation at 30⁰C. Insertion of gene constructs was con- firmed by colony PCR, using gene and construct specific primers found in table 1. For colony PCR, yeast colonies were resuspended in 50 µL 20mM NaOH and incubated at 99⁰C for 15 min.1µL colony suspension was used for PCR. Table 1: List of primers used Assembly of genetic constructs for S. cerevisiae genome engineering Plasmid names and encoded gene constructs are listed in table 2. All plasmids were generated by USER cloning as previously described (5). Also, parent vectors named assembler -1, -2 and -3, for simultaneous genome integration of up to six gene con- structs, and harboring AsiSI/Nb.BsmI USER-cassettes, were prepared for USER cloning as previously described (6). Primers used for PCR amplification with USER compatible PfuX7 polymerase (7) are listed in table 1. Vectors used and generated in this work is listed in table 3. Codon optimized genes for S. cerevisiae were acquired from TWIST Biosciences, USA, San Francisco. All genes denoted with the prefix “CO_” in the below tables were codon- optimized. Codon-optimized genes were amplified using primers identical to those de- scribed in Table 1 above, except that the primers were modified to accommodate hybrid- ization to any nucleotide changes in the codon-optimized genes. The primers for ampli- fication of the codon-optimized genes are also disclosed in J. Andersen-Ranberg et al., Expanding the Landscape of Diterpene Structural Diversity through Stereochemically Controlled Combinatorial Biosynthesis. Angewandte Chemie International Edition, n/a (2016). Table 2: Vectors and plasmids generated and used _

Table 3: List of S. cerevisiae strains used and generated. Name Genotype Source MATα, SUC2, gal2, mal2, mel, National Collection of S288c flo1, flo8-1, ho, bio1, bio6 Yeast Cultures (NCYC) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, NVJ0 ho, bio1, bio6, ura3∆::KanMX, XI-2::(pTEF1- CO_CftTPS1/pPGK1-CO_CftTPS3/pICL1- This study CO_SpGGPPS7/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, XI-2::(pTEF1- NVJ1-3.5 CO_CftTPS1/pPGK1-CO_CftTPS3/pICL1- This study CO_SpGGPPS7), X-4::(pTDH3- TwCYP82D274v1/pTEF2-TwCPR1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, XI-2::(pTEF1- CO_CftTPS1/pPGK1-CO_CftTPS3/pICL1- NVJ3.10 CO_SpGGPPS7), X-4::(pTDH3- This study TwCYP82D274v1/pTEF2-TwCPR1), X- 3::(pTDH3-CO_TwCYP71BE86/pCCW12- CO_TwCYP71BE85v1/pFBA1-Twb5#1) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, XI-2::(pTEF1- CO_CftTPS1/pPGK1-CO_CftTPS3/pICL1- CO_SpGGPPS7), X-4::(pTDH3- NVJ2-19 TwCYP82D274v1/pTEF2-TwCPR1), X- This study 3::(pTDH3-CO_TwCYP71BE86/pCCW12- CO_TwCYP71BE85v1/pENO2- CO_TwCYP82D213/pSED1- TwCYP82D217/pFBA1-TwB5#1/KlURA3) NVJ11-0 MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(KlURA3) This study MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, NVJ11-1 ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- This study CO_CftTPS1/pTEF1-CO_CftTPS3/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- NVJ11-2 TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- This study CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- TwCYP82D274V1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- NVJ11-3 TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- CO_CftTPS1/pTEF1-CO_CftTPS3/pENO2- This study CO_TwCYP71BE85v1/pSED1- TwCYP82D274V1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- NVJ11-4 TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- This study CO_TwCYP71BE86/pSED1- TwCYP82D274V1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- NVJ11-5 CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- This study CO_TwCYP71BE86/pENO2- CO_TwCYP71BE85v1/pSED1- TwCYP82D274V1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- NVJ11-6 CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- CO_TwCYP71BE86/pENO2- This study CO_TwCYP71BE85v1/pPDC1- CO_TwCYP82D213/pSED1- TwCYP82D274V1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- NVJ11-7 TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- This study CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- TwCYP82D274V1/pFBA1-Twb5#1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- NVJ11-8 TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- CO_CftTPS1/pTEF1-CO_CftTPS3/ pENO2- This study CO_TwCYP71BE85v1/pSED1- TwCYP82D274V1/pFBA1-Twb5#1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- NVJ11-9 TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- This study CO_TwCYP71BE86/pSED1- TwCYP82D274V1/pFBA1-Twb5#1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- NVJ11-10 CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- This study CO_TwCYP71BE86/pENO2- CO_TwCYP71BE85v1/pSED1- TwCYP82D274V1/pFBA1-Twb5#1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- NVJ11-11 CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- CO_TwCYP71BE86/pENO2- This study CO_TwCYP71BE85v1/pPDC1- CO_TwCYP82D213/pSED1- TwCYP82D274V1/pFBA1-Twb5#1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- TwCPR2/pICL1-CO_SpGGPPS7/pPGK1- CO_CftTPS1/pTEF1-CO_CftTPS3/pCCW12- CO_TwCYP82D213/pSED1- CO_TwCYP71BE86/pENO2- CO_TwCYP71BE85v1/pPDC1- TwCYP82D213/pSED1-TwCYP82D274V1) NVJ8-15 XI-2::(pPGK1-TwCPR1/pTPI1-Twb5#4) XII- 2::(pPGK1-CO_TwCYP71BE86/pTPI1- This study TwCYP71BE85v1/pTDH3- CO_TwCYP82D213/pSED1- CO_TwCYP82D213/pPDC1- TwCYP82D274V1/pENO2- TwCYP82D274V1/pTEF1- TwCYP82D213/pPGK1- TwCYP82D213/pSED1-Twb5#1/pFBA1- Twb5#4/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- NVJ10-1 CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- CO_TwCYP71BE86/pENO2- This study CO_TwCYP71BE85v1/pPDC1- CO_TwCYP82D213/pSED1- TwCYP82D274v1/pFBA1-Twb5#1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- NVJ10-3 CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- CO_TwCYP71BE86/pENO2- This study CO_TwCYP71BE85v1/pPDC1- CO_TwCYP82D213/pSED1- TwCYP82D274v3/pFBA1-Twb5#1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- NVJ10-6 CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- CO_TwCYP71BE86/pENO2- This study CO_TwCYP71BE85v1/pPDC1- CO_TwCYP82D213/pSED1- TwCYP82D274v4/pFBA1-Twb5#1/KlURA3) MATα, SUC2, gal2, mal2, mel, flo1, flo8-1, ho, bio1, bio6, ura3∆::KanMX, X-3::(pTEF2- TwCPR1/pICL1-CO_SpGGPPS7/pPGK1- NVJ10-8 CO_CftTPS1/pTEF1-CO_CftTPS3/pSED1- CO_TwCYP71BE86/pENO2- This study CO_TwCYP71BE85v1/pPDC1- CO_TwCYP82D213v2/pSED1- TwCYP82D274V1/pFBA1-Twb5#1/KlURA3) Example 3: Expression in the yeast, Saccharomyces cerevisiae Extraction and metabolite analysis Genetically engineered S. cerevisiae strains were transferred into 0.5mL media in a 96- well plate and grown for 3 days at 30 ^C with orbital shaking at 350 rpm. For extraction 0.1mL of S. cerevisiae culture was transferred to 1.5mL glass vials. 0.4mL MetOH uHPLC grade was added. S. cerevisiae extracts extract was filtered by using a 0:22 μm 96-well filter plate (Merck Millipore, Darmstadt, Germany) and at stored at 4 ^C prior to LC-MS analysis. LCMS profiles of the extracts can be seen in figure 2, where it can be observed that transforming the background strain with TwCYP82D274V1 encoding the enzyme having the amino acid sequence of SEQ ID NO: 1 leads to formation of 14-OH-dehydroabietadi- ene. Example 4: Detection of 14-OH-dehydroabietadiene by NMR analysis The compound identified as 14-OH-dehydroabietadiene in example 3 was analyzed by NMR to confirm its identity. Purification for NMR Purification of triptolide intermediates from engineered yeast Engineered yeast producing the desired compound of interest was inoculated from SC- Agar in 10mL YDP and grown ON at 30 ^C.5 mL ON culture was inoculated in 500mL FIT media and grown for 5 days at 30 ^C. Compound of interest was extracted from cul- ture with 500mL EtAc. Solvent was removed by rotor evaporation and analytes were resuspended in hexane. Extraction was repeated 3 times. Hexane extract was applied on Supelclean™ Florisil®/Na2SO4 SPE Tube (Sigma-Aldrich) and analytes were eluted from column using a step gradient with 1:99-5:95 EtAc:Hexane. Each fraction was ana- lyzed with either LC-MS or GC-MS and the fraction containing the compound of interest was selected for NMR analysis. NMR analysis NMR data were acquired on a Bruker Avance III HD 600 MHz NMR spectrometer (1H operating frequency 599.85 MHz) equipped with a 5 mm cryogenically cooled DCH probe optimized for 13C and 1H (Bruker Biospin, Karlsruhe, Germany). NMR data was rec- orded in 5 mm tubes in CDCl3 (Euriso-top, 99.8 atom % D) with temperature equilibration to 300 K, optimization of lock parameters, gradient shimming, and setting of receiver gain, all automatically controlled by Topspin ver. 3.2 and IconNMR ver. 4.7.5 (Bruker Biospin, Karlsruhe, Germany).1H and 13C chemical shifts were referenced to the resid- ual solvent signals of at respectively pH 7.26 ppm and pC 77.16 ppm.1D 1H and 13C NMR spectra were acquired with 30° pulses and 64k data points and zero-filled to 256k data points, 1H spectra were acquired with a spectral width of 12 kHz, a relaxation delay of 1 s and an acquisition time of 2.7 s.13C spectra were 1H-decoupled using the Waltz- 16 composite pulse decoupling scheme.2D homo- and heteronuclear experiments were acquired with 4096 (HMBC), 2048 (DQF-COSY and ROESY), or 1024 (multiplicity edited HSQC) data points in the direct dimension and 256 (DQF-COSY, HMBC and ROESY) or 128 (multiplicity edited HSQC) data points in the indirect dimension. 2D NMR data was zero-filled to 1k in F1 and zero-filled to twice the number of points in F2, employing forward linear prediction in F1 (LPBIN=0). Processing of NMR data was done using Top- spin ver.4.0.9 (Bruker Biospin, Karlsruhe, Germany). The NMR spectroscopic data for 14-OH-dehydroatietadiene is shown in table 4. The ¹ H NMR spectrum of 14-OH-dehydroatietadiene in CDCl ₃ at 599.85 MHz is shown in figure 3 and the ¹³ C NMR spectrum of 14-OH-dehydroatietadiene in CDCl ₃ at 150.83 MHz is shown in figure 4 confirming the identity of the compound. Example 5: Expression in S. cerevisiae of genes leading to production of tripto- phenolide and triptonide This was a preliminary study to assess the effects of expression of genes leading to production of triptophenolide and triptonide in S. cerevisiae. The background yeast strain generated in example 2, was further transformed with vec- tors each containing individual diterpene biosynthesis genes or Tripterygium wilfordii CYPs (TwCYPs) LCMS profiles of the extracts can be seen in figure 5, where it can be seen that trans- forming the background strain with TwCYP82D274V1, TwCYP71BE85 and TwCYP71BE86 encoding the enzymes having the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 3 and SEQ ID NO: 4, respectively; leads to formation of triptophenolide; and further transformation with CYP82D213 encoding the enzyme having the amino acid sequence of SEQ ID NO: 5, leads to the formation of triptonide. Figure 6 shows an overview of the content of oxygenated diterpenoid compounds de- tected in the extracts of the transformants generated. The left panel shows the content of triptophenolide and triptonide, and the right panel shows the content of 14-OH-dehydroabietadiene. Expressing the gene TwB5#1 encod- ing the enzyme having the amino acid sequence of SEQ ID NO: 8 resulted in a signifi- cantly higher production of triptophenolide and triptonide. The genes TwB5#2-6 are other T.wilfordii cytochrome B5 genes (sequences not pro- vided) that do not increase the production of triptophenolide or triptonide. Example 6: Production of oxygenated diterpenoid compounds in S. cerevisiae and N. benthamiana All engineered S. cerevisiae strains and N. benthamiana were cultured as described herein above in the section “Materials and Methods”. Similarly, diterpenoid metabolites were extracted, analyzed by LC-MS, GC-MS and NMR, and quantified as also described herein above. It is preferred that the experimental organism is yeast and that the heterologous genes have been stably transfected in the organism as this gives the most precise and repro- ducible results. The results are shown in Figures 7-9. As can be clearly seen from the figures, organisms as different as yeast cells and tobacco plants are both capable of producing the claimed key intermediates in the proposed biosynthetic pathway of triptonide at high titers ac- cording to the methods of the invention. NMR spectra of other key compounds also produced are shown in Figures 10-26. The NMR spectroscopic data for the produced compounds are shown in Tables 5-21, below.

ROESY H _{-1 ^, H-3} H-1 ^, H-2, H-11, H-20 H _{-1 ^, H-19, H-20 H-1 ^ ^ ^H-5} 10, C-18, C- H-3, H-7A, H-18A H-5 H _{-1 ^ ^ ^ ^- ^ ^} H-15, H-16, H-17 17 H-12 H-12 H-12 H-5 H-2, H-20 H _{-1 ^ ^ ^H-2, H-11, H-19} ROESY _{H-1 ^, H-2B, H-5 H-1 ^, H-11, H-20} H-19B _{H-1 ^, H-19A H-1 ^, H-6 ^, H-7B} H-5, H-7B ^H-20 _{H-6 ^, H5 H-1 ^ ^ ^ ^- ^ ^} H-16, H-17 H-12 H-12 H-2B, H-1' H-2A, H-1' _{H-1 ^ ^ ^H-6 ^, H-11} H-19 H-19 ROESY H _{-1 ^, H-3} H-1 ^, H-2, H-20 H _{-1 ^, H-20 H-1 ^, H-5, H-18A} C-20 H-3, H-7B, H-18A H-7B, H-18A, H-18B H ^{-7B, H-19} H _{-7B, H-6 ^ H-5, H-6 ^, H-6 ^, H-7A} H-15, H-16/17 H-12 H-12 H-12 H _{-3, H-5, H-6 ^, H-19 H-6 ^, H-19 H-6 ^, H-18A, H-18B, H-20 H-1 ^, H-2, H-19} ) ROESY H ^{-3 H-2, H-11, H-20} H _{-1 ^ H-1 ^ ^ ^H-5, H-18} , C-19, C- H-1 ^ ^ ^H-3, H-7 ^, H-18 H _{-7 ^ ^ ^H-7 ^, H-18, H-19} H-7 ^, H-19 H _{-5, H-6 ^ H-6 ^ ^ ^H-6 ^ H-1 ^} H-16, H-17 H-12 H-12 H _{-3, H-5, H-6 ^ H-6 ^ ^ ^H-6 ^, H-20 H-1 ^ ^ ^H-19} ROESY H-3 ^{H-2, H-20} _{H-1 ^, H-19 H-1 ^, H-18 H-3, H-6 ^, H-7B, H-7A} ^{H-5, H-7B, H-18 H-7B, H-19} _{H-5, H-6 ^ H-5, H-6 ^, H-6 ^} H-16/17 H-12 H-12 _{H-3, H-6 ^ H-2, H-6 ^, H-20 H-1 ^, H-19} ROESY H _{-1 ^, H-2B, H-5} ^{18), C-20} _{H-1 ^, H-11, H-20} H-20 H _{-1 ^, H-20 H-1 ^, H-6 ^} H-5, H-6 ^, H-7A, H-7B, H- 19 H-6 ^, H-7B, H-19, H-20 H-6 ^ H _{-6 ^, H-6 ^ H-1 ^} H-16, H-17 H-12 H-12 _{H-6 ^, H-6 ^ H-1 ^, H-2A, H-2B, H-6 ^} ROESY ^{H-5, H-18 H-2, H-11, H-20} _{H-1 ^, H-5, H-18, H-20 H-1 ^ ^ ^H-2, H-7 ^ H-7 ^ ^ ^H-7 ^, H-19A} H-7 ^, H-19A, H-20 _{H-5, H-6 ^ ^ H-6 ^ ^ ^H-7 ^} H-6 ^ ^ ^H-6 ^, H-7 ^ _{H-1 ^} H-16/17 H-12 H-12 _{H-1 ^ ^ ^H-2, H-19B H-6 ^ ^ ^H-6 ^} H-18 H _{-1 ^, H-2, H-6 ^} ROESY H-2B H-3, H-17, H-18 H-1A H-1B, H-5, H-18, C-20 H-3, H-18 H-19 H-19 20 H-14, H-20 H-12A, H-12B, H-17 H-12A, H-12B, H-20 H-11A, H-11B H-11A, H-11B, H-15B H-9 H-15B, H-16 H-12B, H-15A H-7A, H-7B, H-15A, H-17 H-1B, H-11A, H-16 H-1B, H-3, H-5, H-6B H-6A, H-6B H-9, H-11B ROESY H-1 ^, H-5 H-1 ^, H-11, H-20 H-18A, H-18B _{H-1 ^} H-7A, H-7B, H-19 ^{H-7B, H-20} H-6 ^, 14-OH H-6 ^, H-6 ^, 14-OH _{H-1 ^} H-16, H-17 _{H-7 ^ ^ ^H-7 ^, H-15} 14-OH H-12 H-12 H-2, H-19 H-2, H-19 H-6 ^, H-18A, H-18B H _{-1 ^, H-6 ^} ROESY _{H1 ^, H-3} H1 ^, H-2 ^, H-11, H-20 ^{H-18A, H-20} H1 ^, H-18B, H-19 _{H1 ^, H-4, H-18A, H-18B H-3, H-5, H-6 ^ ^ ^H-18B H-4, H-7 ^ H-4, H-7 ^ ^ ^H-7 ^} H-7 ^, 14-OH, H-19, H-20 _{H-5, H-6 ^, 14-OH H-6 ^ ^ ^H-6 ^ H-1 ^, H-20} H-16, H-17 _{H-7 ^ ^ ^H-7 ^, H-15} 14-OH H-12 H-12 _{H-2 ^, H-2 ^, H-3, H-19 H-2 ^, H-3, H-4, H-19 H-2 ^, H-6 ^, H-18B, H-20 H-1 ^, H-2 ^, H-6 ^, H-11, H-19} ROESY _{H1 ^, H-18} H1 ^, H-11, H-20 ^{H-18 H-19, H-20} _{H-6 ^, H-18 H-4, H-7 ^ ^ ^H-7 ^} H-7 ^, 14-OH, H-19, H-20 _{H-6 ^, 14-OH H-6 ^ ^ ^H-6 ^ H-1 ^} H-16, H-17 H-7 ^ ^ ^H-7 ^, H-15 14-OH H-12 H-12 _{H-1 ^, H-2 ^, H-4 H-2 ^, H-6 ^, H-20 H-1 ^, H-2 ^, H-6 ^, H-19} ROESY H _{-2 ^} H-11, H-20 H _{-1 ^, H-19} H ^-20 H _{-7 ^ H-7 ^} H-7 ^, H-20 H-5, H-6 ^, 14-OH H _{-6 ^, 14-OH H-1 ^} H-16, H-17 _{H-7 ^ ^ ^H-7 ^, H-15} 14-OH H-12 H-12 H-19 _{H-2 ^, H-18 H-1 ^, H-2 ^, H-6 ^} ROESY H-1 ^, H-5 H-1 ^, H-2, H-11, H-20B _{H-1 ^, H-18A, H-18B, H-} 20B _{H-1 ^} H-7A, H-7B, H-19 ^{H-7B, H-20A, H-20B} _{H-6 ^, 14-OH} H-6 ^, H-6 ^, 14-OH C-8, C-10, C-13 _{H-1 ^} C-9, C-14, C-15 H-16/17 C-8, C-13 _{H-7A ^ ^H-7B, H-15} C-12, C-13, C-14, C-16, C-17 14-OH C-13, C-15, C-17 H-12 C-13, C-15, C-16 H-12 C-2, C-4 H-2, H-19 C-2, C-4 H-2, H-19 C-3, C-4, C-5 _{H-6 ^, H-18A, H-18B H-6 ^} C-9 _{H-1 ^, H-2, H-6 ^} Table ^Pos. ROESY H _{1 ^, H-3 H1 ^, H-11, H-20} 2 H-18A, H-18B, H-19B, H-20 ³ _{H1 ^, H-4, H-18B} ⁴ _{H-3, H-5, H-6 ^, H-19A H-4, H-7 ^ H-4, H-7 ^ ^ ^H-7 ^, H-19A} H-7 ^, 14-OH, H-19A, H-20 H-5, H-6 ^, 14-OH H _{-6 ^ ^ ^H-6 ^ H-1 ^} H-16, H-17 O _{H-7 ^ ^ ^H-7 ^, H-15} 14-OH H-12 H-12 H-2, H-19B H-2, H-3, H-19B H-4, H-6 ^, H-6 ^, H-20 H-2, H-18A/B, H-20 ²⁰ _{H-1 ^, H-2, H-6 ^, H-19A, H-19B} Pos ROESY . H ^{-3 H-20} H-19 H _{-1 ^, H-5, H-18} H-3, H-7 ^, H-9, H-18 H _{-7 ^, H-18} H-7 ^, H-19 H ^-5 H-6 ^, H-6 ^, H-17B H-5, H-12B, H-14, H-17A, H-20 H-12B, H-14, H-17A, H-20 12 C-15 H-14 H-9, H-11A, H-14, H-15, H-17A H-9, H-11A, H-12A, H-12B H-12B, H-16 H-15 H-9, H-11A, H-12A, H-12B, H-17B H _{-7 ^, H-17A H-3, H-5, H-6 ^ H-2A, H-6 ^} H-9, H-11A 5 ROESY H-3-Me H-4 H-1 H-2 H-12B, H-13 C-14 H-14 C-14 H-10, H-14 H-10 H-12A, H-12B, H-15-Me, H-15-Me' H-14 H-14 ROESY H-3-Me H-4 H-1, H-4 H-2, H-3-Me H-8 H _{-7-Me, H-10, H-15- ^Me} H-11-Me 5 H-8, H-14 H _{-9B, H-12B, H-13A, H-15- ^Me} H-11-Me H-11-Me H-14 H _{-10, H-13B, H-15- ^Me} H ^{-8, H-14 H-11-Me a 1} H NMR (600.13) and ¹³ C NMR (150.90 MHz) data obtained with samples in CDCl ₃ . b Assignments based on HSQC and HMBC experiments. c Multiplicities reported as apparent splittings: s = singlet, d = doublet, t = triplet, sext = sextet, m = multiplet (incl. apping resonances), br = broad. ^ ^denotes Me pointing into the plane and ^ ^denotes Me pointing out of the plane. A denotes owest chemical shift value and B denotes the highest chemical shift value

References 1. J. Andersen-Ranberg et al., Expanding the Landscape of Diterpene Structural Diversity through Stereochemically Controlled Combinatorial Biosynthesis. Angewandte Chemie International Edition, n/a (2016). 2. I. Pateraki et al., Total biosynthesis of the cyclic AMP booster forskolin from Coleus forskohlii. Elife 6, e23001 (2017). 3. I. Pateraki et al., Manoyl Oxide (13R), the Biosynthetic Precursor of Forskolin, Is Synthesized in Specialized Root Cork Cells in Coleus forskohlii. Plant Physiology 164, 1222-1236 (2014). 4. N. L. Hansen et al., The terpene synthase gene family in Tripterygium wilfordii harbors a labdane-type diterpene synthase among the monoterpene synthase TPS-b subfamily. The Plant Journal 89, 429-441 (2017). 5. H. H. Nour-Eldin, B. G. Hansen, M. H. H. Nørholm, J. K. Jensen, B. A. Halkier, Advancing uracil-excision based cloning towards an ideal technique for cloning PCR fragments. Nucleic Acids Research 34, e122 (2006). 6. N. B. Jensen et al., EasyClone: method for iterative chromosomal integration of multiple genes in Saccharomyces cerevisiae. FEMS Yeast Research 14, 238- 248 (2014). 7. M. H. H. Nørholm, A mutant Pfu DNA polymerase designed for advanced uracil- excision DNA engineering. BMC Biotechnology 10, 21 (2010). 8. R. D. Gietz, R. H. Schiestl, High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method. Nature Protocols 2, 31-34 (2007). 9. Hansen, N.L., et al., Integrating pathway elucidation with yeast engineering to produce polpunonic acid the precursor of the anti-oDesity agent celastrol. MicroD Cell Fact, 2020.19(1): p.15. 10. Voinnet O, Rivas S, Mestre P, Baulcombe D. An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus [retracted in: Plant J.2015 Nov;84(4):846]. Plant J. 2003;33(5):949-956. doi:10.1046/j.1365-313x.2003.01676.x ITEMS 1. A recombinant host cell, capable of producing oxygenated diterpenoid compound, wherein the host cell is capable of producing miltiradiene and/or dehydroabietadiene and has been transformed with a first gene en- coding an enzyme having cytochrome P450 activity and which enzyme is capable of converting miltiradiene and/or dehydroabietadiene into 14-OH- dehydroabietadiene. 2. The recombinant host cell of item 1, wherein the first gene encodes a pol- ypeptide comprising an amino acid sequence having at least 80 % se- quence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 1 (TwCYP82D274V1) or the mature polypeptide thereof. 3. The recombinant host cell of item 1 or 2, wherein the recombinant host cell further comprises: a second gene encoding a second enzyme having cytochrome P450 activity and a third gene encoding a third enzyme having cytochrome P450 activity, wherein: the second gene encoding an enzyme having cytochrome P450 activity encodes a polypeptide comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% se- quence identity, preferably at least 90% sequence identity, prefera- bly at least 95% sequence identity, more preferred at least 98% se- quence identity to SEQ ID NO: 4 (TwCYP71BE86) or the mature pol- ypeptide thereof; the third gene encoding an enzyme having cytochrome P450 ac- tivity encodes a polypeptide comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% se- quence identity, preferably at least 90% sequence identity, prefera- bly at least 95% sequence identity, more preferred at least 98% se- quence identity to SEQ ID NO: 3 (TwCYP71BE85) or the mature pol- ypeptide thereof. 4. The recombinant host cell of item 3, wherein the host cell further comprises a gene encoding a polypeptide having cytochrome B5 activity and com- prising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% se- quence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 8 (TwB5#1) or the mature polypeptide thereof. 5. The recombinant host cell of item 3 or 4, wherein the host cell further com- prises a fourth gene encoding a fourth enzyme having cytochrome P450 activity; wherein: the fourth gene encoding a fourth enzyme having cytochrome P450 ac- tivity encodes a polypeptide comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% se- quence identity more preferred at least 98% sequence identity to SEQ ID NO: 5 (TwCYP82D213) or the mature polypeptide thereof. 6. The recombinant host cell of item 5, wherein the host cell further comprises a fifth gene encoding a fifth enzyme having cytochrome P450 activity and/or a sixth gene encoding a sixth enzyme having cytochrome P450 activity, wherein: the fifth gene encoding a fifth enzyme having cytochrome P450 activity encodes a polypeptide comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, prefer- ably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 6 (TwCYP82D217) or the mature polypeptide thereof ; and the sixth gene encoding a sixth enzyme having cytochrome P450 activity encodes a polypeptide comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, prefer- ably at least 90% sequence identity, preferably at least 95% sequence identity, more preferred at least 98% sequence identity to SEQ ID NO: 7 (TwCYP82D275) or the mature polypeptide thereof . 7. The recombinant host cell according to any of the preceding items, wherein the host cell capable of producing miltiradiene and/or dehydro- abietadiene is a recombinant cell that has been transformed with one or more gene(s) encoding: a. a geranylgeranyl diphosphate synthase; b. a diterpene synthase capable of converting GGPP into miltiradiene; c. a combination of two or more diterpene synthases that in combina- tion is capable of converting GGPP into miltiradiene; or d. a copalyl diphosphate synthase and a miltiradiene synthase. 8. The recombinant host cell of item 7, wherein the geranylgeranyl diphos- phate synthase is a polypeptide comprising the amino acid sequence of SEQ ID NO: 73 or SEQ ID NO: 81. 9. The recombinant host cell of item 7, wherein the combination of two or more diterpene synthases, that is capable of converting GGPP into milti- radiene, is the combination of a polypeptide comprising the amino acid sequence of SEQ ID NO: 67 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 68; or is the combination of a polypeptide com- prising the amino acid sequence of SEQ ID NO: 69 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 70. 10.The recombinant host cell of item 7, wherein the combination of a copalyl diphosphate synthase and a miltiradiene synthase is a combination of a polypeptide comprising the amino acid sequence of SEQ ID NO: 71 and a polypeptide comprising the polypeptide of SEQ ID NO: 72. 11.The recombinant host cell according to any of the previous items wherein the host cell is selected among prokaryotic and eukaryotic cells. 12.The recombinant host cell according to item 11, being a prokaryotic cell selected among Escherichia, Bacillus, Lactobacillus and Corynebacterium species. 13.The recombinant host cell of item 11, being a eukaryotic cell selected among Saccharomyces, Scizosaccharomýces, Klyveromyces, Pichia, Candida and Yarrowia species. 14.The recombinant host cell of item 11, where the cell is a S. cerevisiae cell. 15.Use of a recombinant host cell according to any of the preceding items for the production of an oxygenated diterpenoid compound. 16.The use of item 15, wherein the oxygenated diterpenoid compound is se- lected among 14-OH-dehydroabietadiene, triptophenolide and triptonide. 17.The use of item 16, wherein the oxygenated diterpenoid compound is trip- tonide, which triptonide is further converted into triptolide. 18.The use according to one of the items 15-17, wherein the oxygenated diterpenoid compound is recovered using one or more separation and/or chromatographic steps. 19.A polypeptide having cytochrome P450 enzyme activity and comprising an amino acid sequence having at least 80 % sequence identity, preferably at least 85% sequence identity, preferably at least 90% sequence identity, preferably at least 95% sequence identity, preferably at least 96% sequence identity, preferably at least 97% sequence identity, preferably at least 98% sequence identity, or even 100% sequence identity to one of the sequences SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 or the mature polypeptide thereof. 20.A polynucleotide encoding the polypeptide of item 19. 21.A plasmid, expression vector, expression construct or recombinant host cell comprising a polynucleotide of item 20. 22.The compound 14-OH-dehydroabietadiene.