Production of DHA and other LC PUFAs in plants

Patent metadata

Field	Value
Publication number	US-11053511-B2
Application number	US-201514726285-A
Country	US
Kind code	B2
Filing date	May 29, 2015
Priority date	May 17, 2010
Publication date	Jul 6, 2021
Grant date	Jul 6, 2021

Abstract

The invention provides 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 present invention also relates to methods of making and using such organisms as well as products obtained from such organisms.

First claim

What is claimed is:

1. A genetically modified soybean plant or seed thereof, comprising:
a first polynucleotide operably linked to a seed-specific promoter, the polynucleotide encoding a polyunsaturated fatty acid (PUFA) synthase polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO: 1, wherein the first polynucleotide comprises a nucleotide sequence that is at least 90% identical to SEQ ID NO:6;
a second polynucleotide operably linked to a seed-specific promoter, the polynucleotide encoding a PUFA synthase system polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:2, wherein the second polynucleotide comprises a nucleotide sequence that is at least 90% identical to SEQ ID NO:7;
a third polynucleotide operably linked to a seed-specific promoter, the polynucleotide encoding a PUFA synthase polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:3, wherein the third polynucleotide comprises a nucleotide sequence that is at least 90% identical to SEQ ID NO:8;
a fourth polynucleotide operably linked to a seed-specific promoter, the polynucleotide encoding a phosphopantetheinyl transferase (PPTase) polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:5, wherein the fourth polynucleotide comprises a nucleotide sequence that is at least 90% identical to SEQ ID NO: 10; and
a fifth polynucleotide encoding an acyl-CoA synthetase (ACoAS) comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:4, wherein seed of the soybean plant comprises an oil comprising docosahexaenoic acid (C22:6, n-3) (DHA) and docosapenaenoic acid (C22:5, n-6) (DPA(n-6)).

2. The genetically modified soybean plant or seed thereof of claim 1 , wherein the first polynucleotide encodes the PUFA synthase polypeptide of SEQ ID NO:1, wherein the second polynucleotide encodes the PUFA synthase polypeptide of SEQ ID NO:2, and wherein the third polynucleotide encodes the PUFA synthase polypeptide of SEQ ID NO:3.

3. The genetically modified soybean plant or seed thereof of claim 2 , wherein the first polynucleotide comprises the nucleotide sequence of SEQ ID NO:6, wherein the second polynucleotide comprises the nucleotide sequence of SEQ ID NO:7, and wherein the third polynucleotide comprises the nucleotide sequence of SEQ ID NO:8.

4. The genetically modified soybean plant or seed thereof of claim 1 , wherein the fourth polynucleotide encodes the PPTase of SEQ ID NO:5.

5. The genetically modified soybean plant or seed thereof of claim 1 , wherein each of the polynucleotides is operably linked to the PvDlec2 promoter.

6. The genetically modified soybean plant or seed thereof of claim 1 , wherein the fifth polynucleotide encodes the ACoAS of SEQ ID NO:4.

7. The genetically modified soybean plant or seed thereof of claim 1 , further comprising a polynucleotide encoding an acetyl CoA carboxylase (ACCase) or a polynucleotide encoding a type 2 diacylglycerol acyltransferase (DGAT2).

8. The genetically modified soybean plant or seed thereof of claim 1 , wherein the oil comprises 0.7%-12% combined docosahexaenoic acid (C22:6, n-3) (DHA)+docosapentaenoic acid (C22:5, n-6) (DPA(n-6)) by weight of total fatty acids.

9. The genetically modified soybean plant or seed thereof of claim 8 , wherein the oil comprises 0.7%-2.8% DHA by weight of total fatty acids.

10. The genetically modified soybean plant or seed thereof of claim 8 , wherein the oil comprises 0.5%-1.9% DPA(n-6) by weight of total fatty acids.

11. The genetically modified soybean plant or seed thereof of claim 9 , wherein the oil comprises 0.5%-1.9% DPA(n-6) by weight of total fatty acids.

12. The genetically modified soybean plant or seed thereof of claim 8 , wherein the plant or seed thereof is a soybean seed.

13. A food product comprising the soybean seed of claim 12 .

14. A functional food comprising the genetically modified soybean plant or seed thereof of claim 8 .

15. A pharmaceutical product comprising the genetically modified soybean plant or seed thereof of claim 8 .

16. The genetically modified soybean plant or seed thereof of claim 8 ,
wherein the PUFA synthase polypeptide encoded by the first polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:28 or SEQ ID NO:29 at its amino terminus,
wherein the PUFA synthase polypeptide encoded by the second polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:28 or SEQ ID NO:29 at its amino terminus,
wherein the PUFA synthase polypeptide encoded by the third polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:28 or SEQ ID NO:29 at its amino terminus, and
wherein the PPTase encoded by the fourth polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:28 or SEQ ID NO:29 at its amino terminus.

17. The genetically modified soybean plant material or seed thereof of claim 16 ,
wherein the PUFA synthase polypeptide encoded by the first polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:29 at its amino terminus,
wherein the PUFA synthase polypeptide encoded by the second polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:29 at its amino terminus,
wherein the PUFA synthase polypeptide encoded by the third polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:29 at its amino terminus, or
wherein the PPTase encoded by the fourth polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:29 at its amino terminus.

18. A genetically modified soybean seed comprising:
a first polynucleotide operably linked to a seed-specific promoter, the polynucleotide comprising the nucleotide sequence of SEQ ID NO:6;
a second polynucleotide operably linked to a seed-specific promoter, the polynucleotide comprising the nucleotide sequence of SEQ ID NO:7;
a third polynucleotide operably linked to a seed-specific promoter, the polynucleotide comprising the nucleotide sequence of SEQ ID NO:8;
a fourth polynucleotide operably linked to a seed-specific promoter, the polynucleotide comprising the nucleotide sequence of SEQ ID NO:10; and
a fifth polynucleotide encoding the acyl-CoA synthetase (ACoAS) of SEQ ID NO:4,
wherein the plant seed comprises a seed oil with a polyunsaturated fatty acid profile consisting essentially of 0.36%-8.00% docosahexaenoic acid (C22:6, n-3) (DHA) and 0.3%-3.98% docosapentaenoic acid (C22:5, n-6) (DPA(n-6)) by weight of total fatty acids.

19. The genetically modified soybean seed of claim 8 , wherein the fifth polynucleotide consists of a nucleotide sequence at least 90% identical to SEQ ID NO:9.

20. The genetically modified soybean seed of claim 8 , wherein the fifth polynucleotide consists of a nucleotide sequence at least 90% identical to SEQ ID NO:30.

21. The genetically modified soybean seed of claim 20 ,
wherein the PUFA synthase polypeptide encoded by the first polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:28 or SEQ ID NO:29 at its amino terminus,
wherein the PUFA synthase polypeptide encoded by the second polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:28 or SEQ ID NO:29 at its amino terminus,
wherein the PUFA synthase polypeptide encoded by the third polynucleotide comprises the chloroplast transit peptide (CTP) encoded by SEQ ID NO:28 or SEQ ID NO:29 a

Assignees

• Dow Agrosciences Llc
• Dsm Ip Assets Bv

Inventors

• Walsh Terence A
• Gachotte Daniel
• Merlo Ann Owens
• Pareddy Dayakar Reddy
• Metz James
• Bevan Scott
• Kuner Jerry

Classifications

• C12N15/8243 Primary

  involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine · CPC title

• C12N15/8247 Primary

  involving modified lipid metabolism, e.g. seed oil composition · CPC title

• A23L11/03

  Soya beans, e.g. full-fat soya bean flakes or grits · CPC title

• C12Y203/01085

  Fatty-acid synthase (2.3.1.85) · CPC title

• A01H6/542 Primary

  Glycine max [soybean] · CPC title