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Title:
NUCLEIC ACID MOLECULES ENCODING ALTERNANSUCRASE
Document Type and Number:
WIPO Patent Application WO/2000/047727
Kind Code:
A2
Abstract:
Nucleic acid molecules encoding an alternansucrase are provided. Moreover, vectors, host cells and plant cells transformed by the herein-described nucleic acid molecules and plants containing them are provided. Furthermore, methods are described for preparing transgenic plants which synthesize the carbohydrate alternan, because of the insertion of nucleic acid molecules encoding an alternansucrase. Moreover, methods for preparing alternan and products resulting from them are provided.

Inventors:
KOSSMANN JENS (DE)
WELSH THOMAS (DE)
QUANZ MARTIN (DE)
KNUTH KAROLA (DE)
Application Number:
PCT/EP2000/000954
Publication Date:
August 17, 2000
Filing Date:
February 07, 2000
Export Citation:
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Assignee:
PLANTTEC BIOTECHNOLOGIE GMBH (DE)
MAX PLANCK GESELLSCHAFT (DE)
KOSSMANN JENS (DE)
WELSH THOMAS (DE)
QUANZ MARTIN (DE)
KNUTH KAROLA (DE)
International Classes:
A23L1/09; A61K8/60; A61K8/73; A61Q19/00; C08B37/00; C12N1/21; C12N5/10; C12N9/10; C12N15/31; C12N15/54; C12N15/09; C12P19/02; C12P19/04; C12P19/08; C12P19/18; C12R1/19; (IPC1-7): C12N9/10; C12N15/54; C12N15/63; C12N15/70; C12N15/82; C12N5/10; A01H5/00; C12P21/02; C12P19/04; C12P19/18
Domestic Patent References:
WO1995013389A11995-05-18
WO1996004365A11996-02-15
Other References:
None
Attorney, Agent or Firm:
VOSSIUS & PARTNER (Siebertstrasse 4 München, DE)
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Claims:
Claims
1. A nucleic acid molecule encoding a protein having the biological activity of an alternansucrase, selected from the group consisting of (a) nucleic acid molecules encoding at least the mature form of a protein comprising the amino acid sequence indicated in Seq. ID No. 2 or the amino acid sequence which is encoded by the cDNA contained in plasmid DSM 12666; (b) nucleic acid molecules comprising the nucleotide sequence indicated in Seq. ID No. 1 or the nucleotide sequence of the cDNA contained in plasmid DSM 12666 or a corresponding ribonucleotide sequence; (c) nucleic acid molecules encoding a protein, the amino acid sequence of which has a homology of at least 40% to the amino acid sequence indicated in Seq. ID No 2; (d) nucleic acid molecules, one strand of which hybridizes with any one of the nucleic acid molecules as defined in (a) or (b); (e) nucleic acid molecules comprising a nucleotide sequence encoding a biologically active fragment of the protein which is encoded by any one of the nucleic acid molecules as defined in (a), (b), (c) or (d); and (f) nucleic acid molecules, the nucleotide sequence of which deviates on account of the degeneration of the genetic code from the sequence of the nucleic acid molecules as defined in (a), (b), (c), (d) or (e).
2. An oligonucleotide or polynucleotide which specifically hybridizes with a nucleic acid molecule of claim 1.
3. A vector containing a nucleic acid molecule according to claim 1.
4. The vector according to claim 3, wherein the nucleic acid molecule is connected in sense orientation to regulatory elements ensuring the transcription and synthesis of a translatable RNA in prokaryotic or eukaryotic cells.
5. Plasmid pAlsupSK deposited under the accession No. DSM 12666.
6. A host cell transformed with a nucleic acid molecule of claim 1 or a vector of claim 3 or 4 or descended from such a cell.
7. The host cell according to claim 6, which is a cell of a microorganism.
8. The host cell according to claim 6, which is an E. coli cell.
9. A method for preparing a protein having the biological activity of an alternansucrase or a biologically active fragment thereof, wherein a host cell of any one of claims 6 to 8 is cultured under conditions permitting the synthesis of the protein, and wherein the protein is isolated from the cultured cells and/or the culture medium.
10. A protein or biologically active fragment thereof encoded by a nucleic acid molecule of claim 1 or prepared according to the method of claim 9.
11. A transgenic plant cell transformed with a nucleic acid molecule of claim 1 or a vector of claim 3 or 4, or descended from such a cell, wherein said nucleic acid molecule encoding the protein having the biological activity of an alternansucrase is under the control of regulatory elements permitting the transcription of a translatable mRNA in plant cells.
12. A plant containing the plant cells of claim 11.
13. The plant according to claim 12, which is a useful plant.
14. The plant according to claim 12 or 13, which is a sugarstoring or starchstoring plant.
15. Propagation material of a plant according to any one of claims 12 to 14, containing the plant cells of claim 11.
16. A method for preparing alternan comprising the steps of extracting and isolating the alternan from a plant according to any one of claims 12 to 14.
17. Alternan obtainable from a plant according to any one of claims 12 to 14, from propagation material according to claim 15 or by a method according to claim 16.
18. A method for preparing alternan and/or fructose, wherein (a) a host cell of any one of claim 6 to 8 secrets an alternansucrase into a saccharosecontaining culture medium; and (b) alternan and/or fructose is/are isolated from the culture medium.
19. The method according to claim 18, wherein the host cell is immobilized.
20. A method for preparing alternan and/or fructose, wherein (a) a saccharosecontaining solution is contacted with a protein of claim 10 under conditions permitting the conversion of saccharose into alternan and/or fructose; and (b) alternan and/or fructose is/are isolated from the solution.
21. The method according to claim 20, wherein the protein is immobilized on a carrier material.
22. A method for preparing alternan and/or fructose, wherein (a) a saccharosecontaining solution is contacted with a protein of claim 10 and acceptor molecules under conditions permitting the conversion of saccharose into alternan and/or fructose; and (b) alternan and/or fructose is/are isolated from the solution.
23. The method according to claim 22, wherein the acceptor molecule is selected from the group consisting of maltose, isomaltose, isomaltotriose and methyla Dglucan.
24. The method according to claim 22 or 23, wherein the protein is immobilized.
25. Cosmetic products or food products containing alternan as specified in claim 17 or prepared according to any one of claims 16 and 18 to 24.
26. A method for preparing cosmetic products or food products comprising a process according to any of claims 16 and 18 to 24 and the formulation of the resulting alternan in a form suitable for use as a cosmetic product or foodstuff.
Description:
INTERNATIONAL SEARCH REPORT lnteri nal AppileMlon No PCT/EP 00/00954 C. (Continuation) DOCUMENTS CONSIDERED TO BE RELEVANT Category Citation of document, with indication, where appropriate, of the relevant passages Relevant to daim No. A RAEMAEKERS M H M ET AL:"PRODUCTION OF 1,17,20, ALTERNANSUCRASE BY LEUCONOSTOC 26 MESENTEROIDES NRRL B-1355 IN BATCH FERMENTATION WITH CONTROLLED PH AND DISSOLVE OXYGEN" JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY. (INTERNATIONAL JOURNAL OF BIOTECHNICAL AND CHEMICAL PROCESSES), GB, ELSEVIER APPLIED SCIENCE PUBLISHERS. BARKING, vol. 69, no. 4, 1 August 1997 (1997-08-01), pages 470-478, XP000702350 ISSN: 0268-2575 abstract page 471, left-hand column P, X ARGUELLO-MORALES ET AL.:"Sequence 1-10 analysis of the gene encoding alternansucrase, a sucrose glucosyltransferase from Leuconostoc mesenteroides NRRL B-1355" FEMS MICROBIOLOGY LETTERS, vol. 182,1 January 2000 (2000-01-01), pages 81-85, XP000937860 abstract 1 INTERNATIONAL SEARCH REPORT star, emppucatb to . nformation on patent fam lly membws PCT/EP 00/00954 Patent document Publication Patent family Publication cited in search report date member (s) date WO 9513389 A 18-05-1995 AU 1208795 A 29-05-1995 BR 9408286 A 26-08-1997 CA 2176109 A 18-05-1995 EP 0728213 A 28-08-1996 HU 75075 A 28-04-1997 JP 9505467 T 03-06-1997 PL 314296 A 02-09-1996 US 5908975 A 01-06-1999 ZA 9408867 A 09-05-1996 WO 9604365 A 15-02-1996 AU 697819 B 15-10-1998 AU 3235295 A 04-03-1996 CA 2196596 A 15-02-1996 EP 0804545 A 05-11-1997 US 5702942 A 30-12-1997 US 5789209 A 04-08-1998



 
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