Delta-8 desaturase and its use in making polyunsaturated fatty acids

• US 20060195939A1
• Filed: 06/24/2005
• Published: 08/31/2006
• Est. Priority Date: 06/25/2004
• Status: Active Grant

• Current Assignee: Corteva Agriscience LLC (Corteva, Inc.)
• Original Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Inventors: Damude, Howard Glenn, Zhu, Quinn Qun
• Granted Patent: US 7,550,651 B2
• Time in Patent Office: Days
• US Class Current: 800/281
• CPC Class Codes: A23D 9/00 Other edible oils or fats, e.g. shortenings, cooking oils
• CPC Class Codes: A23K 20/158 Fatty acids; Fats; Products containing oils or fats
• CPC Class Codes: A23L 33/12 Fatty acids or derivatives thereof
• CPC Class Codes: A23V 2002/00 Food compositions, function of food ingredients or processes for food or foodstuffs
• CPC Class Codes: A23V 2300/21 Genetic modification
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/0083 Miscellaneous (1.14.99)
• CPC Class Codes: C12P 7/6427 Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
• CPC Class Codes: C12P 7/6431 Linoleic acids [18:2[n-6]]
• CPC Class Codes: C12P 7/6432 Eicosapentaenoic acids [EPA]
• CPC Class Codes: C12P 7/6434 Docosahexenoic acids [DHA]
• CPC Class Codes: C12P 7/6472 containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone

Abstract

Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding a delta-8 desaturase along with a method of making long chain polyunsaturated fatty acids (PUFAs) using this delta-8 desaturase in plants and oleaginous yeast.

Claims

1. An isolated polynucleotide comprising:

(a) a nucleotide sequence encoding a polypeptide having delta-8 desaturase activity, wherein the polypeptide has an amino acid sequence consisting essentially of SEQ ID NOs;

2 or 113;

or,

(b) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary.

2. The polynucleotide of claim 1 wherein the nucleotide sequence comprises SEQ ID NOs:

1 or 112.

3. The polypeptide of claim 1, wherein the amino acid sequence of the polypeptide comprises SEQ ID NOs:

2 or 113.

4. A recombinant construct comprising the polynucleotide of claim 1 operably linked to at least one regulatory sequence.

5. A cell comprising the recombinant construct of claim 4.

6. The cell of claim 5 wherein said cell is selected from the group consisting of plants and yeast.

7. A transformed Yaffowia sp. comprising the recombinant construct of claim 4.

8. The transformed Yarrowia sp. of claim 7 selected from the group consisting of Yarrowia lipolytica ATCC #20362, Yarrowia lipolytica ATCC #8862, Yarrowia lipolytica ATCC #18944, Yarrowia lipolytica ATCC #76982 and Yarrowia lipolytica LGAM S(7)1.

9. A method for transforming a cell, comprising transforming a cell with the recombinant construct of claim 4 and selecting those cells transformed with the recombinant construct of claim 4.

10. A method for producing a transformed plant comprising transforming a plant cell with the polynucleotide of claim 1 and regenerating a plant from the transformed plant cell.

11. The method of claim 10 wherein the plant is a soybean plant.

12. A method for producing yeast comprising transforming a yeast cell with the polynucleotide of claim 1 and growing yeast from the transformed yeast cell.

13. A method for producing yeast comprising transforming a yeast cell with the polynucleotide of claim 1 and growing yeast from the transformed yeast cell, wherein the yeast is an oleaginous yeast selected from the group of:

Yarrowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon and Lipomyces.

14. A seed comprising the recombinant construct of claim 4.

15. A seed obtained from the plant made by the method of of claim 10 or 11.

16. A method for making long chain polyunsaturated fatty acids in a cell comprising:

(a) transforming a cell with the recombinant construct of claim 4;

(b) selecting those transformed cells that make long chain polyunsaturated fatty acids.

17. Oil obtained from the seed of claim 14.

18. Oil obtained from the seed of claim 15.

19. Oil obtained from the yeast made by the method of claim 12 or 13.

20. A method for making long chain polyunsaturated fatty acids in a plant cell comprising:

(a) transforming a cell with the recombinant construct of claim 4;

(b) selecting those transformed cells that make long chain polyunsaturated fatty acids.

22. An oilseed plant comprising the recombinant construct of claim 4.

24. The plant of claim 22 or 23, wherein the oilseed plant is selected from the group consisting of soybean, Brassica species, sunflower, maize, cotton, flax, and safflower.

25. The plant of claim 22 or 23 wherein the oilseed plant is selected from the group consisting of soybean, Brassica species, sunflower, maize, cotton, flax, and safflower and further wherein the polyunsaturated fatty acid is selected from the group consisting of AA, EDA, EPA, ETA, EtrA, DGLA, DPA and DHA.

26. A seed obtained from the plant of claim 22.

28. Oil obtained from the seed of claim 26.

30. Oil obtained by the method of claim 20 or 21

31. Food or feed which incorporates the oil of claim 28.

33. Food or feed which incorporates the oil of claim 30.

34. Food or feed comprising the seeds of claims 31.

36. Food or feed comprising the seeds of claims 33.

37. Food or feed comprising an ingredient derived from the processing of the seeds of claim 26.

21. A method for producing at least one polyunsaturated fatty acid in a soybean cell comprising:

(a) transforming a soybean cell with a first recombinant DNA construct comprising an isolated polynucleotide encoding a delta-8 desaturase polypeptide, operably linked to at least one regulatory sequence and at least one additional recombinant DNA construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide selected from the group consisting of a delta-4, a delta-5, delta-6, a delta-9, a delta-12, a delta-15, and a delta-17 desaturase, a delta-9 elongase, a C18 to C22 elongase and a C20 to C24 elongase.

(b) regenerating a soybean plant from the transformed cell of step (a); and

(c) selecting those seeds obtained from the plants of step (b) having an altered level of polyunsaturated fatty acids when compared to the level in seeds obtained from a nontransformed soybean plant.

23. An oilseed plant comprising:

a) a first recombinant DNA construct comprising an isolated polynucleotide encoding a delta-8 desaturase polypeptide, operably linked to at least one regulatory sequence; and

b) at least one additional recombinant DNA construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide selected from the group consisting of a delta-4, a delta-5, delta-6, a delta-9, a delta-12, a delta-1 5, and a delta-17 desaturase, a delta-9 elongase, a C18 to C22 elongase and a C20 to C24 elongase.

27. A seed obtained from the plant of claim 23.

29. Oil obtained from the seed of claim 27.

32. Food or feed which incorporates the oil of claim 29.

35. Food or feed comprising the seeds of claims 32.

38. Food or feed comprising an ingredient derived from the processing of the seeds of claim 27.