RU2014102130A - Мультизимы и их использование в получении полиненасыщенных жирных кислот - Google Patents
Мультизимы и их использование в получении полиненасыщенных жирных кислот Download PDFInfo
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- 235000020777 polyunsaturated fatty acids Nutrition 0.000 title 1
- 239000002157 polynucleotide Substances 0.000 claims abstract 14
- 102000040430 polynucleotide Human genes 0.000 claims abstract 14
- 108091033319 polynucleotide Proteins 0.000 claims abstract 14
- 239000002773 nucleotide Substances 0.000 claims abstract 12
- 125000003729 nucleotide group Chemical group 0.000 claims abstract 12
- 101000912235 Rebecca salina Acyl-lipid (7-3)-desaturase Proteins 0.000 claims abstract 4
- 101000877236 Siganus canaliculatus Acyl-CoA Delta-4 desaturase Proteins 0.000 claims abstract 4
- 230000002255 enzymatic effect Effects 0.000 claims abstract 4
- 229920001184 polypeptide Polymers 0.000 claims abstract 4
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract 4
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 230000001105 regulatory effect Effects 0.000 claims abstract 2
- 210000004027 cell Anatomy 0.000 claims 20
- 241000196324 Embryophyta Species 0.000 claims 14
- MBMBGCFOFBJSGT-KUBAVDMBSA-N all-cis-docosa-4,7,10,13,16,19-hexaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 claims 4
- 230000001131 transforming effect Effects 0.000 claims 3
- 210000005253 yeast cell Anatomy 0.000 claims 3
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 claims 2
- 235000020669 docosahexaenoic acid Nutrition 0.000 claims 2
- 229940090949 docosahexaenoic acid Drugs 0.000 claims 2
- 235000020673 eicosapentaenoic acid Nutrition 0.000 claims 2
- 229960005135 eicosapentaenoic acid Drugs 0.000 claims 2
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 claims 2
- 244000068988 Glycine max Species 0.000 claims 1
- 244000020551 Helianthus annuus Species 0.000 claims 1
- 241000235013 Yarrowia Species 0.000 claims 1
- 230000000813 microbial effect Effects 0.000 claims 1
- 230000001172 regenerating effect Effects 0.000 claims 1
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- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
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- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
- C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
- C12N15/8247—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified lipid metabolism, e.g. seed oil composition
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
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- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12P7/6434—Docosahexenoic acids [DHA]
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- C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
- C12Y114/19—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with oxidation of a pair of donors resulting in the reduction of molecular oxygen to two molecules of water (1.14.19)
- C12Y114/19001—Stearoyl-CoA 9-desaturase (1.14.19.1), i.e. DELTA9-desaturase
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Abstract
1. Мультизим, включающий отдельный полипептид, обладающий, по меньшей мере, двумя независимыми и раздельными ферментативными активностями, где указанные ферментативные активности представляют собой дельта-4-десатуразу и C-элонгазу, и где указанные дельта-4-десатураза и C-элонгаза связаны связью, выбранной из группы, состоящей из SEQ ID NO:198, SEQ ID NO:200, SEQ ID NO:235, SEQ ID NO:438, SEQ ID NO:472, SEQ ID NO:445 и SEQ ID NO:504.2. Выделенный полинуклеотид, кодирующий мультизим по п. 1, где указанный полинуклеотид включает:(a) нуклеотидную последовательность, кодирующую полипептид, имеющий аминокислотную последовательность, которая, по меньшей мере, на 80% идентична SEQ ID NO:12, SEQ ID NO:22, SEQ ID NO:95, SEQ ID NO:96 или SEQ ID NO:97;(b) нуклеотидную последовательность, приведенную в SEQ ID NO:11, SEQ ID NO:205, SEQ ID NO:21, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93 или SEQ ID NO:410; или(c) нуклеотидную последовательность, которая гибридизуется в жестких условиях с нуклеотидной последовательностью, приведенной в SEQ ID NO:11, SEQ ID NO:205, SEQ ID NO:21, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93 или SEQ ID NO:410.3. Выделенный полинуклеотид по п. 2, где указанный полинуклеотид включает нуклеотидную последовательность, кодирующую SEQ ID NO:12, SEQ ID NO:22, SEQ ID NO:95, SEQ ID NO:96 или SEQ ID NO:97.4. Выделенный полинуклеотид по п. 3, где указанныйполинуклеотид включает нуклеотидную последовательность, приведенную в SEQ ID NO:11, SEQ ID NO:205, SEQ ID NO:21, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93 или SEQ ID NO:410.5. Рекомбинантная конструкция, включающая любой из выделенных полинуклеотидов по пп. 2-4, функционально связанный, по меньшей мере, с одной регуляторной последовательностью.6. Клетка-хозяин, содержащая рекомбинантную конструкцию по п. 5.7. Клетка-хозяин по п. 6, где указанная клетка хозяин представляет собой клетку растения.8. Клетка-х�
Claims (18)
1. Мультизим, включающий отдельный полипептид, обладающий, по меньшей мере, двумя независимыми и раздельными ферментативными активностями, где указанные ферментативные активности представляют собой дельта-4-десатуразу и C20/22-элонгазу, и где указанные дельта-4-десатураза и C20/22-элонгаза связаны связью, выбранной из группы, состоящей из SEQ ID NO:198, SEQ ID NO:200, SEQ ID NO:235, SEQ ID NO:438, SEQ ID NO:472, SEQ ID NO:445 и SEQ ID NO:504.
2. Выделенный полинуклеотид, кодирующий мультизим по п. 1, где указанный полинуклеотид включает:
(a) нуклеотидную последовательность, кодирующую полипептид, имеющий аминокислотную последовательность, которая, по меньшей мере, на 80% идентична SEQ ID NO:12, SEQ ID NO:22, SEQ ID NO:95, SEQ ID NO:96 или SEQ ID NO:97;
(b) нуклеотидную последовательность, приведенную в SEQ ID NO:11, SEQ ID NO:205, SEQ ID NO:21, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93 или SEQ ID NO:410; или
(c) нуклеотидную последовательность, которая гибридизуется в жестких условиях с нуклеотидной последовательностью, приведенной в SEQ ID NO:11, SEQ ID NO:205, SEQ ID NO:21, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93 или SEQ ID NO:410.
3. Выделенный полинуклеотид по п. 2, где указанный полинуклеотид включает нуклеотидную последовательность, кодирующую SEQ ID NO:12, SEQ ID NO:22, SEQ ID NO:95, SEQ ID NO:96 или SEQ ID NO:97.
4. Выделенный полинуклеотид по п. 3, где указанный
полинуклеотид включает нуклеотидную последовательность, приведенную в SEQ ID NO:11, SEQ ID NO:205, SEQ ID NO:21, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93 или SEQ ID NO:410.
5. Рекомбинантная конструкция, включающая любой из выделенных полинуклеотидов по пп. 2-4, функционально связанный, по меньшей мере, с одной регуляторной последовательностью.
6. Клетка-хозяин, содержащая рекомбинантную конструкцию по п. 5.
7. Клетка-хозяин по п. 6, где указанная клетка хозяин представляет собой клетку растения.
8. Клетка-хозяин по п. 7, где указанная клетка растения представляет собой клетку масличного растения.
9. Клетка-хозяин по п. 6, где указанная клетка-хозяин представляет собой клетку дрожжей.
10. Клетка-хозяин по п. 9, где указанная клетка-хозяин представляет собой клетку жировых дрожжей.
11. Клетка-хозяин по п. 10, где указанная клетка дрожжей представляет собой клетку Yarrowia.
12. Способ трансформирования клетки, включающий трансформирование клетки рекомбинантной конструкцией по п. 5 и отбор клеток, трансформированных указанной рекомбинантной конструкцией.
13. Способ получения трансформированного растения, включающий трансформирование клетки растения рекомбинантной конструкцией по п. 5 и регенерирование растения из указанной трансформированной клетки растения.
14. Растение, содержащее рекомбинантную конструкцию по п. 5.
15. Растение по п. 14, где указанное растение представляет собой масличное растение.
16. Растение по п. 15, где указанное растение представляет собой растение сои или растение подсолнечника.
17. Семя, полученное из растения по п. 14, где указанное семя содержит указанную рекомбинантную конструкцию.
18. Способ превращения эйкозапентаеновой кислоты в докозагексаеновую кислоту, включающий:
a) обеспечение рекомбинантной микробной клетки-хозяина, содержащей рекомбинантную конструкцию по п. 5 и источник эйкозапентаеновой кислоты; и
b) выращивание указанной клетки-хозяина из (a) в условиях, в которых образуется докозагексаеновая кислота.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US90979007P | 2007-04-03 | 2007-04-03 | |
| US60/909,790 | 2007-04-03 | ||
| US2789808P | 2008-02-12 | 2008-02-12 | |
| US61/027,898 | 2008-02-12 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| RU2009140397/10A Division RU2517608C2 (ru) | 2007-04-03 | 2008-04-03 | Мультизимы и их использование в получении полиненасыщенных жирных кислот |
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| RU2014102130A true RU2014102130A (ru) | 2015-08-27 |
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| RU2009140397/10A RU2517608C2 (ru) | 2007-04-03 | 2008-04-03 | Мультизимы и их использование в получении полиненасыщенных жирных кислот |
| RU2014102130/10A RU2014102130A (ru) | 2007-04-03 | 2014-01-23 | Мультизимы и их использование в получении полиненасыщенных жирных кислот |
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| RU2009140397/10A RU2517608C2 (ru) | 2007-04-03 | 2008-04-03 | Мультизимы и их использование в получении полиненасыщенных жирных кислот |
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| Country | Link |
|---|---|
| US (2) | US8828690B2 (ru) |
| EP (1) | EP2129777B1 (ru) |
| JP (1) | JP2010523113A (ru) |
| CN (2) | CN101765658B (ru) |
| BR (1) | BRPI0808577A2 (ru) |
| CA (1) | CA2679988A1 (ru) |
| DK (1) | DK2129777T3 (ru) |
| ES (1) | ES2559312T3 (ru) |
| HU (1) | HUE028692T2 (ru) |
| MX (1) | MX2009010574A (ru) |
| RS (1) | RS20090413A (ru) |
| RU (2) | RU2517608C2 (ru) |
| UA (1) | UA103595C2 (ru) |
| WO (1) | WO2008124048A2 (ru) |
Families Citing this family (48)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102559364B (zh) | 2004-04-22 | 2016-08-17 | 联邦科学技术研究组织 | 用重组细胞合成长链多不饱和脂肪酸 |
| ES2715640T3 (es) | 2004-04-22 | 2019-06-05 | Commw Scient Ind Res Org | Síntesis de ácidos grasos poliinsaturados de cadena larga por células recombinantes |
| CA2661697A1 (en) | 2006-08-29 | 2008-03-06 | Commonwealth Scientific And Industrial Research Organisation | Synthesis of fatty acids |
| US7794701B2 (en) * | 2007-04-16 | 2010-09-14 | E.I. Du Pont De Nemours And Company | Δ-9 elongases and their use in making polyunsaturated fatty acids |
| US8957280B2 (en) * | 2007-05-03 | 2015-02-17 | E. I. Du Pont De Nemours And Company | Delta-5 desaturases and their use in making polyunsaturated fatty acids |
| EP2176416B1 (de) * | 2007-07-31 | 2016-03-16 | BASF Plant Science GmbH | Elongasen und verfahren zur herstellung mehrfach ungesättigter fettsäuren in transgenen organismen |
| CN101970638B (zh) | 2007-10-03 | 2015-02-11 | 纳幕尔杜邦公司 | 用于高水平生产二十碳五烯酸的优化解脂耶氏酵母菌株 |
| EP2274429B1 (en) | 2008-04-03 | 2014-10-22 | E. I. du Pont de Nemours and Company | Multizymes |
| US8168858B2 (en) | 2008-06-20 | 2012-05-01 | E. I. Du Pont De Nemours And Company | Delta-9 fatty acid elongase genes and their use in making polyunsaturated fatty acids |
| BR122021003836B1 (pt) | 2008-11-18 | 2022-02-01 | Commonwealth Scientific And Industrial Research Organisation | Polinucleotídeo isolado e/ou exógeno que não ocorre de forma natural, vetor e método para produzir óleo contendo ácidos graxos insaturados |
| AU2009335903B2 (en) | 2008-12-18 | 2016-04-14 | E. I. Du Pont De Nemours And Company | Reducing byproduction of malonates in a fermentation process |
| US8524485B2 (en) * | 2009-06-16 | 2013-09-03 | E I Du Pont De Nemours And Company | Long chain omega-3 and omega-6 polyunsaturated fatty acid biosynthesis by expression of acyl-CoA lysophospholipid acyltransferases |
| CA2765911C (en) | 2009-06-16 | 2018-12-18 | E.I. Du Pont De Nemours And Company | High eicosapentaenoic acid oils from improved optimized strains of yarrowia lipolytica |
| BRPI1009649A2 (pt) | 2009-06-16 | 2016-03-15 | Du Pont | célula hospedeira yarrowia sp, recombinante, método para fabricar um óleo microbiano e célula hospedeira recombinante para produção de um óleo |
| JP5931721B2 (ja) | 2009-06-16 | 2016-06-08 | イー・アイ・デュポン・ドウ・ヌムール・アンド・カンパニーＥ．Ｉ．Ｄｕ Ｐｏｎｔ Ｄｅ Ｎｅｍｏｕｒｓ Ａｎｄ Ｃｏｍｐａｎｙ | アシルＣｏＡリゾリン脂質アシルトランスフェラーゼの発現による長鎖オメガ−３及びオメガ−６多価不飽和脂肪酸生合成の改良 |
| WO2011008510A2 (en) | 2009-06-30 | 2011-01-20 | E. I. Du Pont De Nemours And Company | Plant seeds with altered storage compound levels, related constructs and methods involving genes encoding cytosolic pyrophosphatase |
| CA3037072C (en) | 2009-07-17 | 2022-07-26 | Basf Plant Science Company Gmbh | Novel fatty acid desaturases and elongases and uses thereof |
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| AR078829A1 (es) | 2009-10-30 | 2011-12-07 | Du Pont | Plantas y semillas con niveles alterados de compuesto de almacenamiento, construcciones relacionadas y metodos relacionados con genes que codifican proteinas similares a las aldolasas bacterianas de la clase ii del acido 2,4- dihidroxi-hept-2-eno-1,7-dioico |
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| BR112012022045A8 (pt) | 2010-03-03 | 2018-06-19 | Du Pont | ''planta transgênica,semente transgênica,método para produzir uma planta transgênica,método para produzir sementes transgênicas,produto e/ou subproduto,polinucleotídeo isolado e planta ou semente''. |
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| WO2004090123A2 (de) * | 2003-04-08 | 2004-10-21 | Basf Plant Science Gmbh | Δ-4-desaturasen aus euglena gracilis, exprimierende pflanzen und pufa enthaltende öle |
| US7238482B2 (en) * | 2003-05-07 | 2007-07-03 | E. I. Du Pont De Nemours And Company | Production of polyunsaturated fatty acids in oleaginous yeasts |
| US7214491B2 (en) * | 2003-05-07 | 2007-05-08 | E. I. Du Pont De Nemours And Company | Δ-12 desaturase gene suitable for altering levels of polyunsaturated fatty acids in oleaginous yeasts |
| US7125672B2 (en) | 2003-05-07 | 2006-10-24 | E. I. Du Pont De Nemours And Company | Codon-optimized genes for the production of polyunsaturated fatty acids in oleaginous yeasts |
| EP2172536B1 (de) * | 2003-08-01 | 2016-05-04 | BASF Plant Science GmbH | Verfahren zur Herstellung mehrfach ungesättigter Fettsäuren in transgenen Organismen |
| US7504259B2 (en) | 2003-11-12 | 2009-03-17 | E. I. Du Pont De Nemours And Company | Δ12 desaturases suitable for altering levels of polyunsaturated fatty acids in oleaginous yeast |
| AU2004290051A1 (en) | 2003-11-12 | 2005-05-26 | E.I. Dupont De Nemours And Company | Delta-15 desaturases suitable for altering levels of polyunsaturated fatty acids in oilseed plants and oleaginous yeast |
| EP3543324B1 (de) * | 2004-02-27 | 2022-11-30 | BASF Plant Science GmbH | Verfahren zur herstellung mehrfach ungesättigten fettsäuren in transgenen pflanzen |
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- 2008-04-03 RU RU2009140397/10A patent/RU2517608C2/ru not_active IP Right Cessation
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- 2008-04-03 MX MX2009010574A patent/MX2009010574A/es active IP Right Grant
- 2008-04-03 HU HUE08727277A patent/HUE028692T2/en unknown
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- 2008-04-03 UA UAA200910197A patent/UA103595C2/ru unknown
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| US20150031096A1 (en) | 2015-01-29 |
| CA2679988A1 (en) | 2008-10-16 |
| WO2008124048A8 (en) | 2011-11-03 |
| WO2008124048A3 (en) | 2009-02-05 |
| HUE028692T2 (en) | 2016-12-28 |
| CN101765658A (zh) | 2010-06-30 |
| UA103595C2 (ru) | 2013-11-11 |
| ES2559312T3 (es) | 2016-02-11 |
| RS20090413A (sr) | 2010-06-30 |
| CN104152423A (zh) | 2014-11-19 |
| WO2008124048A2 (en) | 2008-10-16 |
| EP2129777B1 (en) | 2015-10-28 |
| EP2129777A2 (en) | 2009-12-09 |
| MX2009010574A (es) | 2009-10-22 |
| CN101765658B (zh) | 2013-11-20 |
| US20080254191A1 (en) | 2008-10-16 |
| RU2517608C2 (ru) | 2014-05-27 |
| JP2010523113A (ja) | 2010-07-15 |
| AU2008236723A1 (en) | 2008-10-16 |
| US8828690B2 (en) | 2014-09-09 |
| RU2009140397A (ru) | 2011-05-10 |
| BRPI0808577A2 (pt) | 2019-09-24 |
| DK2129777T3 (en) | 2016-01-25 |
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