Production of omega 3 long chain polyunsaturated fatty acids in oilseed crops by a thraustochytrid PUFA synthase

Patent metadata

Field	Value
Publication number	US-9994828-B2
Application number	US-201414555326-A
Country	US
Kind code	B2
Filing date	Nov 26, 2014
Priority date	Nov 26, 2013
Publication date	Jun 12, 2018
Grant date	Jun 12, 2018

Abstract

This disclosure concerns recombinant host organisms genetically modified with a polyunsaturated fatty acid (PUFA) synthase system and one or more accessory proteins that allow for and/or improve the production of PUFAs in the host organism. The disclosure also concerns methods of making and using such organisms as well as products obtained from such organisms.

First claim

What may be claimed is:

1. A genetically modified Brassica plant that produces oil comprising eicosapentaenoic acid (C20:5, n-3) (EPA) in amount between 0.01% and 10% by weight of total fatty acids, the plant comprising:
a first polynucleotide encoding a first polypeptide of a polyunsaturated fatty acid (PUFA) synthase system comprising SEQ ID NO:1;
a second polynucleotide encoding a second polypeptide of the PUFA system comprising SEQ ID NO:4;
a third polynucleotide encoding a third polypeptide of the PUFA system consisting of SEQ ID NO:14; and
a fourth polynucleotide encoding a phosphopantetheinyl transferase (PPTase) from a Nostoc sp.,
wherein each of the polynucleotides is operably linked to a promoter functional in the plant.

2. A cell, tissue, or part of the genetically modified plant of claim 1 .

3. The genetically modified plant of claim 1 , wherein the promoter is a seed-specific promoter or a leaf-specific promoter.

4. The genetically modified plant of claim 1 , wherein the promoter is a seed-specific promoter, and wherein the polynucleotides are operably linked to a transcription termination sequence.

5. The genetically modified plant of claim 1 , wherein the promoter is selected from the group consisting of PvDlec2; LfKCS3; FAE1; BoACP; BnaNapinC; ubiquitin; CsVMV; SSPRO2745.1; and SSPRO2743.1 promoters.

6. The genetically modified plant of claim 1 , wherein the oil produced by the plant further comprises DHA (docosahexaenoic acid (C22:6, n-3)) in an amount between 0.01% and 15% by weight of total fatty acids.

7. The genetically modified plant of claim 6 , wherein the oil produced by the plant comprises between 0.05% and 10% DHA by weight of total fatty acids.

8. The genetically modified plant of claim 7 , wherein the oil produced by the plant comprises between 0.05% and 5% DHA by weight of total fatty acids.

9. The genetically modified plant of claim 1 , wherein the oil produced by the plant comprises an amount of EPA between 0.05% and 5% by weight of total fatty acids.

10. The genetically modified plant of claim 9 , wherein the oil produced by the plant comprises an amount of EPA between 0.05% and 1% by weight of total fatty acids.

11. The genetically modified plant of claim 1 , wherein the plant or cell, tissue, seed, or part thereof comprises a ratio of EPA:DHA (docosahexaenoic acid (C22:6, n-3)) of between 1:1 and 1:30 by weight of total fatty acids.

12. The genetically modified plant of claim 11 , wherein the oil produced by the plant comprises a ratio of EPA:DHA of between 1:1 and 1:3 by weight of total fatty acids.

13. A seed obtained from the genetically modified plant of claim 1 , the seed comprising the first, second, third, and fourth polynucleotides.

14. A commodity product obtained from the genetically modified plant of claim 1 , wherein the commodity product is selected from the group consisting of a refined oil, an unrefined oil, a raw oil, a feed or meal composition, and a functional food product, the commodity product comprising the first, second, third, and fourth polynucleotides.

15. A method for obtaining oil comprising at least one PUFA, the method comprising recovering oil from the genetically modified plant of claim 1 .

16. The method according to claim 15 , wherein the oil is recovered from a cell, tissue, seed, or part of the genetically modified plant.

17. The genetically modified plant of claim 1 , wherein the first polynucleotide is at least 70% identical to at least one of SEQ ID NO:2 and SEQ ID NO:3, wherein the second polynucleotide is at least 70% identical to at least one of SEQ ID NO:5 and SEQ ID NO:6, and wherein the third polynucleotide is at least 70% identical to SEQ ID NO:13.

18. The genetically modified plant of claim 1 , wherein the first polynucleotide is SEQ ID NO:3, wherein the second polynucleotide is SEQ ID NO:6, and wherein the third polynucleotide is SEQ ID NO:13.

19. The genetically modified plant of claim 1 , wherein the polynucleotides are arranged in a head-to-tail configuration in the same nucleic acid molecule.

20. The genetically modified plant of claim 1 , wherein the first polynucleotide and the fourth polynucleotide are arranged in a tail-to-tail configuration, and wherein the second polynucleotide and the third polynucleotide are arranged in a tail-to-tail configuration in the same nucleic acid molecule.

21. The genetically modified plant of claim 1 , wherein the first polynucleotide and the fourth polynucleotide are arranged in a head-to-head configuration, and wherein the second polynucleotide and the third polynucleotide are arranged in a head-to-head configuration in the same nucleic acid molecule.

22. The genetically modified plant of claim 1 ,
wherein the first polynucleotide hybridizes under stringent conditions to the complement of SEQ ID NO:2 or SEQ ID NO:3,
wherein the second polynucleotide hybridizes under stringent conditions to the complement of SEQ ID NO:5 or SEQ ID NO:6,
wherein the third polynucleotide hybridizes under stringent conditions to the complement of SEQ ID NO:13, or
wherein the fourth polynucleotide hybridizes under stringent conditions to the complement of SEQ ID NO:10.

23. An isolated nucleic acid molecule comprising:
a first polynucleotide encoding a first polypeptide of a polyunsaturated fatty acid (PUFA) synthase system comprising SEQ ID NO:1;
a second polynucleotide encoding a second polypeptide of the PUFA system comprising SEQ ID NO:4;
a third polynucleotide encoding a third polypeptide of the PUFA system consisting of SEQ ID NO:14; and
a fourth polynucleotide encoding the phosphopantetheinyl transferase (PPTase) from a Nostoc sp.,
wherein each of the polynucleotides is operably linked to a promoter functional in the plant.

24. The isolated nucleic acid molecule of claim 23 , wherein the first polynucleotide is at least 70% identical to at least one of SEQ ID NO:2 and SEQ ID NO:3, wherein the second polynucleotide is at least 70% identical to at least one of SEQ ID NO:5 and SEQ ID NO:6, wherein the third polynucleotide is at least 70% identical to SEQ ID NO:13, or wherein the fourth polynucleotide is at least 70% identical to SEQ ID NO:10.

25. The isolated nucleic acid molecule of claim 23 , wherein the first polynucleotide is SEQ ID NO:3, wherein the second polynucleotide is SEQ ID NO:6, wherein the third polynucleotide is SEQ ID NO:13, and wherein the fourth polynucleotide is SEQ ID NO:10.

26. The isolated nucleic acid molecule of claim 23 , wherein the polynucleotides are all arranged in a head-to-tail configuration.

27. The isolated nucleic acid molecule of claim 23 , wherein the first and fourth polynucleotides are arranged in a tail-to-tail configuration, and wherein the second and third polynucleotides are arranged in a tail-to-tail configuration.

28. The isolated nucleic acid molecule of claim 23 , wherein the first and fourth polynucleotides are arranged in a head-to-head configuration, and wherein the second and third polynucleotides are arranged in a head-to-head configuration.

29. A system for producing a genetically modified plant, the system comprising at least one isolated nucleic acid molecules, the system comprising:
a first polynucleotide encoding a first polypeptide of a polyunsaturated fatty acid (PUFA) synthase system comprising SEQ ID NO:1;
A second polynucleotide encoding a second polypeptide

Assignees

• Dow Agrosciences Llc
• Dsm Ip Assets Bv

Inventors

• Walsh Terence A
• Gachotte Daniel J
• Larsen Cory M
• Bevan Scott
• Owens-Merlo P Ann
• Metz James G
• Zirkle Ross

Classifications

• A23D9/00

  Other edible oils or fats, e.g. shortenings or cooking oils (preservation of other edible oils A23B20/30) · CPC title

• C12N9/1029 Primary

  transferring groups other than amino-acyl groups (2.3.1) · CPC title

• C11B1/10

  by extracting · CPC title

• C12Y203/01085

  Fatty-acid synthase (2.3.1.85) · CPC title

• A23K20/158

  Fatty acids; Fats; Products containing oils or fats · CPC title