Method for producing polyunsaturated fatty acids in transgenic plants 机翻标题: 暂无翻译,请尝试点击翻译按钮。

公开号/公开日
US2009222951 A1 2009-09-03 [US20090222951]US9458436 B2 2016-10-04 [US9458436] / 2009-09-032016-10-04
申请号/申请日
2005US-10590457 / 2005-02-23
发明人
CIRPUS PETRA;BAUER JÖRG;QIU XIAO;WU GUOHAI;DATLA NAGAMANI;
申请人
BASF PLANT SCIENCE;
主分类号
IPC分类号
A23D-009/00A61K-008/36A61K-008/92A61K-031/202A61Q-019/00C12N-009/02C12N-009/10C12N-015/82
摘要
(US9458436) The present invention relates to a process for the production of polyunsaturated fatty acids in the seed of transgenic plants by introducing, into the organism, nucleic acids which encode polypeptides with a ω3-desaturase, Δ12-desaturase, Δ6-desaturase, Δ6-elongase, Δ5-desaturase, Δ5-elongase and/or Δ4-desaturase activity.  The invention furthermore relates to recombinant nucleic acid molecules comprising the nucleic acid sequences which encode the aforementioned polypeptides, either jointly or individually, and transgenic plants which comprise the aforementioned recombinant nucleic acid molecules.  Furthermore, the invention relates to the generation of a transgenic plant and to oils, lipids and/or fatty acids with an elevated content of polyunsaturated fatty acids, in particular arachidonic acid, eicosapentaenoic acid and/or docosahexaenoic acid, as the result of the expression of the elongases and desaturases used in the process according to the invention.
机翻摘要
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地址
代理人
代理机构
;
优先权号
2004DE-10009457 2004-02-27 2004DE-10012370 2004-03-13 2004DE-10017518 2004-04-08 2004DE-10024014 2004-05-14 2004DE-10062543 2004-12-24 2004WO-EP07957 2004-07-16 2005WO-EP01863 2005-02-23
主权利要求
(US9458436) We claim: 1.  A process for the production of compounds of the general formula I   in seeds of a transgenic plant with a content of at least 20% by weight based on the total lipid content, which comprises the following process steps:   a) introducing, into a plant, at least one nucleic acid sequence which encodes a polypeptide with DELTA 6-desaturase activity,   b) introducing, into the plant, at least one nucleic acid sequence which encodes a polypeptide with DELTA 6-elongase activity,   c) introducing, into the plant, at least one nucleic acid sequence which encodes a polypeptide with DELTA 5-desaturase activity,   d) introducing, into the plant, at least one nucleic acid sequence which encodes a polypeptide with DELTA 5-elongase activity, wherein said DELTA 5-elongase activity elongates only unsaturated C20-fatty acids, and   e) introducing, into the plant, at least one nucleic acid sequence which encodes a polypeptide with DELTA 4-desaturase activity, and   wherein the variables and substituents in formula I have the following meanings:   R1=hydroxyl, coenzyme A (thioester), lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysodiphosphatidylglycerol, lysophosphatidylserine, lysophosphatidylinositol, sphingo base or a radical of the general formula II    R2=hydrogen, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysodiphosphatidylglycerol, lysophosphatidylserine, lysophosphatidylinositol or saturated or unsaturated C2-C24-alkylcarbonyl,   R3=hydrogen, saturated or unsaturated C2-C24-alkylcarbonyl, or R2 and R3 independently of one another are a radical of the general formula Ia:    in which   n=2, 3, 4, 5, 6, 7 or 9, m=2, 3, 4, 5 or 6 and p=0 or 3,   and wherein the at least one nucleic acid sequence which encodes a polypeptide with DELTA 5-elongase activity comprises:   i) the nucleic acid sequence of SEQ ID NO: 67, 83, or 113;   ii) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 68, 84, or 114;   iii) a nucleic acid sequence having at least 95% identity to the nucleic acid sequence of SEQ ID NO: 67, 83, or 113; or   iv) a nucleic acid sequence encoding an amino acid sequence having at least 95% identity to SEQ ID NO: 68, 84, or 114. 2. The process according to claim 1, wherein the variables n, m and p have the following meanings: n=2, 3 or 5, m=4, 5 or 6 and p=0 or 3. 3. The process according to claim 1, wherein, in formula I, the variables n, m and p have the following meanings: (a) m=4, n=3, p=3 and the compound is arachidonic acid,   (b) m=5, n=3, p=0 and the compound is eicosapentaenoic acid,   (c) m=5, n=5, p=0 and the compound is docosapentaenoic acid, or   (d) m=6, n=3, p=0 and the compound is docosahexaenoic acid. 4. The process according to claim 2, wherein, in the seed of the transgenic plant, the content of all compounds of the formula I together amounts to at least 27% by weight based on the total lipid content. 5. The process according to claim 3, wherein, in the seed of the transgenic plant, the docosahexaenoic acid content amounts to at least 1% by weight based on the total lipid content. 6. The process according to claim 1, wherein the nucleic acid sequences which encode polypeptides with DELTA 6-desaturase, DELTA 6-elongase, DELTA 5-desaturase, or DELTA 4-desaturase activity are selected from the group consisting of: a) a nucleic acid sequence with the sequence shown in SEQ ID NO: 11, SEQ ID NO: 27, SEQ ID NO: 41, or SEQ ID NO: 193,   b) nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO: 12, SEQ ID NO: 28, SEQ ID NO: 42, or SEQ ID NO: 194, and   c) derivatives of the nucleic acid sequence shown in SEQ ID NO: 11, SEQ ID NO: 27, SEQ ID NO: 41, or SEQ ID NO: 193, which encode polypeptides with at least 70% identity at the amino acid level with SEQ ID NO: 12, SEQ ID NO: 28, SEQ ID NO: 42, or SEQ ID NO: 194, and which have DELTA 6-desaturase, DELTA 6-elongase, DELTA 5-desaturase or DELTA 4-desaturase activity. 7. The process according to claim 1, wherein a nucleic acid sequence which encodes polypeptides with omega 3-desaturase activity, selected from the group consisting of: a) a nucleic acid sequence with the sequence shown in SEQ ID NO: 87 or SEQ ID NO: 105, or   b) nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO: 88 or SEQ ID NO: 106, or   c) derivatives of the nucleic acid sequence shown in SEQ ID NO: 87 or SEQ ID NO: 105, which encode polypeptides with at least 70% identity at the amino acid level with SEQ ID NO: 88 or SEQ ID NO: 106 and which have omega 3-desaturase activity   is additionally introduced into the transgenic plant. 8. The process according to claim 1, wherein a nucleic acid sequence which encodes polypeptides with DELTA 12-desaturase activity, selected from the group consisting of: a) a nucleic acid sequence with the sequence shown in SEQ ID NO: 107, SEQ ID NO: 109 or SEQ ID NO: 195, or   b) nucleic acid sequences which, as the result of the degeneracy of the genetic code, can be derived from the amino acid sequence shown in SEQ ID NO: 108, SEQ ID NO: 110 or SEQ ID NO: 196, or   c) derivatives of the nucleic acid sequence shown in SEQ ID NO: 107, SEQ ID NO: 109 or SEQ ID NO: 195, which encode polypeptides with at least 70% identity at the amino acid level with SEQ ID NO: 108, SEQ ID NO: 110 or SEQ ID NO: 196 and which have DELTA 12-desaturase activity   is additionally introduced into the transgenic plant. 9. The process according to claim 1, wherein a nucleic acid sequence which encodes proteins of the biosynthetic pathway of the fatty acid or lipid metabolism selected from the group acyl-CoA dehydrogenase(s), acyl-ACP [=acyl carrier protein] desaturase(s), acyl-ACP thioesterase(s), fatty acid acyltransferase(s), acyl-CoA:lysophospholipid acyltransferase(s), fatty acid synthase(s), fatty acid hydroxylase(s), acetyl-coenzyme A carboxylase(s), acyl-coenzyme A oxidase(s), fatty acid desaturase(s), fatty acid acetylenases, lipoxygenases, triacylglycerol lipases, allenoxide synthases, hydroperoxide lyases or fatty acid elongase(s) is additionally introduced into the transgenic plant. 10. The process according to claim 1, wherein the substituents R2 or R3 independently of one another are saturated or unsaturated C18-C22-alkylcarbonyl. 11. The process according to claim 1, wherein the substituents R2 or R3 independently of one another are unsaturated C18-, C20- or C22-alkylcarbonyl with at least two double bonds. 12. The process according to claim 1, wherein the transgenic plant is selected from the group consisting of an oil-producing plant, a vegetable plant and an ornamental. 13. The process according to claim 1, wherein the transgenic plant is selected from the group of the plant families consisting of: Anacardiaceae, Asteraceae, Boraginaceae, Brassicaceae, Cannabaceae, Compositae, Cruciferae, Cucurbitaceae, Elaeagnaceae, Euphorbiaceae, Fabaceae, Geraniaceae, Gramineae, Leguminosae, Linaceae, Malvaceae, Moringaceae, Marchantiaceae, Onagraceae, Olacaceae, Oleaceae, Papaveraceae, Piperaceae, Pedaliaceae, Poaceae and Solanaceae. 14. The process according to claim 1, wherein the compounds of the general formula I are isolated from the transgenic plant in the form of their oils, lipids or free fatty acids. 15. The process according to claim 1, wherein the polypeptide with DELTA 5-elongase activity elongates only unsaturated C20-fatty acids with one double bond in the DELTA 5-position. 16. The process according to claim 1, wherein the compounds of the general formula I comprise fatty acids having 20 or 22 carbon atoms in the fatty acid chain. 17. The process according to claim 1, wherein the plant is selected from the group consisting of soybean, peanut, oilseed rape, canola, linseed, evening primrose, mullein, thistle, hazelnut, almond, macadamia, avocado, bay, wild roses, pumpkin/squash, pistachios, sesame, sunflower, safflower, borage, maize, poppy, mustard, hemp, castor-oil plant, olive, Calendula, Punica, oil palm, walnut and coconut. 18. A process for the production of arachidonic acid (ARA), eicosapentaenoic acid (EPA) and/or docosahexaenoic acid (DHA) in seeds of a plant, comprising introducing into a plant: a) a nucleic acid encoding a polypeptide having DELTA 6-desaturase activity;   b) a nucleic acid encoding a polypeptide having DELTA 6-elongase activity;   c) a nucleic acid encoding a polypeptide having DELTA 5-desaturase activity;   d) a nucleic acid encoding a polypeptide having DELTA 5-elongase activity; and   e) a nucleic acid encoding a polypeptide having DELTA 4-desaturase activity;   wherein said nucleic acid encoding a polypeptide having DELTA 5-elongase activity comprises:   i) the nucleotide sequence of SEQ ID NO: 67, 83, or 113;   ii) a nucleic acid sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 68, 84, or 114; or   iii) a nucleic acid sequence encoding a polypeptide having at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 68, 84, or 114. 19. The process of claim 18, wherein said nucleic acid encoding a polypeptide having DELTA 5-elongase activity comprises a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 68, 84, or 114. 20. The process of claim 18, wherein said nucleic acid encoding a polypeptide having DELTA 5-elongase activity comprises a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 68, 84, or 114. 21. The process of claim 1, wherein EPA and/or DHA is produced in the seeds of said transgenic plant. 22. A process for the production of an oil-, lipid- and fatty acid-composition, comprising: a) obtaining EPA and/or DHA produced by the process of claim 18; and   b) formulating said EPA and/or DHA as an oil-, lipid- and fatty acid-composition. 23. A method for the production of feedstuffs, foodstuffs, cosmetics or pharmaceuticals, comprising: a) obtaining an oil-, lipid- and fatty acid-composition produced by the process of claim 22; and   b) processing said oil-, lipid- and fatty acid-composition to produce feedstuffs, foodstuffs, cosmetics or pharmaceuticals. 24. A process for the production of an oil-, lipid- and fatty acid-composition, comprising: a) producing EPA and/or DHA in seeds of a transgenic plant according to the process of claim 18; and   b) obtaining an oil-, lipid- and fatty acid-composition from the seeds of said transgenic plant. 25. A method for the production of feedstuffs, foodstuffs, cosmetics or pharmaceuticals, comprising: a) obtaining an oil-, lipid- and fatty acid-composition produced by the process of claim 24; and   b) processing said oil-, lipid- and fatty acid-composition to produce feedstuffs, foodstuffs, cosmetics or pharmaceuticals.
法律状态
(US9458436) LEGAL DETAILS FOR US2009222951  Actual or expected expiration date=2025-02-23    Legal state=ALIVE    Status=GRANTED     Event publication date=2005-02-23  Event code=US/APP  Event indicator=Pos  Event type=Examination events  Application details  Application country=US US10590457  Application date=2005-02-23  Standardized application number=2005US-10590457     Event publication date=2006-10-20  Event code=US/AS  Event type=Change of name or address  Event type=Reassignment  Assignment OWNER: BASF PLANT SCIENCE GMBH, GERMANY ASSIGNMENT OF ASSIGNORS INTEREST ASSIGNORS:CIRPUS, PETRA BAUER, JORG QIU, XIAO AND OTHERS REEL/FRAME:018416/0670 SIGNING DATES FROM 20050308 TO 20050329     Event publication date=2006-10-20  Event code=US/AS  Event type=Change of name or address  Event type=Reassignment  Assignment OWNER: BASF PLANT SCIENCE GMBH, GERMANY ASSIGNMENT OF ASSIGNORS INTEREST ASSIGNORS:CIRPUS, PETRA BAUER, JORG QIU, XIAO AND OTHERS SIGNING DATES FROM 20050308 TO 20050329 REEL/FRAME:018416/0670     Event publication date=2009-09-03  Event code=US/A1  Event type=Examination events  Application published  Publication country=US  Publication number=US2009222951  Publication stage Code=A1  Publication date=2009-09-03  Standardized publication number=US20090222951     Event publication date=2016-10-04  Event code=US/B2  Event indicator=Pos  Event type=Event indicating In Force  Granted patent as second publication  Publication country=US  Publication number=US9458436  Publication stage Code=B2  Publication date=2016-10-04  Standardized publication number=US9458436
专利类型码
A1B2
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