Fatty acid desaturase gene obtained from pomegranate and method for the production of unsaturated fatty acids

• US 20050166271A1
• Filed: 07/09/2002
• Published: 07/28/2005
• Est. Priority Date: 07/20/2001
• Status: Abandoned Application

• Current Assignee: BASF Plant Science GmbH (BASF SE)
• Original Assignee: BASF Plant Science GmbH (BASF SE)
• Inventors: Feubner, Ivo, Pernstich, Christian, Renz, Andreas, Hornung, Ellen
• Application Number:
  US10/484,202
• Publication Number:
  US 20050166271A1
• Time in Patent Office: Days
• US Class Current: 800/8
• 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 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12P 7/6472 containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone

Abstract

The present invention relates to a process for the production of unsaturated or saturated fatty acids and to a process for the production of oils and/or triglycerides with an increased content of unsaturated or saturated fatty acids. The invention furthermore relates to nucleic acid sequences, nucleic acid constructs, vectors and organisms comprising the nucleic acid sequences, nucleic acid constructs and/or vectors. Moreover, the invention relates to fatty acid mixtures and to triglycerides with an increased content of unsaturated fatty acids, and to their use.

Claims

1. An isolated nucleic acid that encodes a polypeptide with a desaturase activity, wherein said nucleic acid contains a sequence selected from the group consisting of:

the sequence of SEQ ID NO;

2;

the sequence of SEQ ID NO;

7;

a sequence derived from the amino acid sequence of SEQ ID NO;

3 or SEQ ID NO;

8 by back translation of the respective amino acid sequences, owing to the degeneracy of the genetic code; and

a derivative of the sequence of SEQ ID NO;

2 or SEQ ID NO;

7 wherein said derivative encodes a peptide that has at least 75 % identity at the amino acid level to said polypeptide, wherein the desaturase activity of the peptide is not substantially reduced as compared with the desaturase activity of said polypeptide.

2. An amino acid sequence encoded by the nucleic acid of claim 1.

3. The amino acid sequence of claim 2, which contains the sequence of SEQ ID NO:

3 or SEQ ID NO;

8.

4. A nucleic acid construct comprising the nucleic acid of claim 1 linked to one or more regulatory signal sequences.

5. A vector comprising the nucleic acid construct of claim 4.

6. An organism comprising the vector of claim 5.

7. The organism of claim 6, which is a plant, a microorganism or an animal.

8. A transgenic plant comprising the nucleic acid of claim 1.

9. A process for production of oils or triglycerides with an increased content of unsaturated fatty acids, which comprises:

introducing the nucleic acid of claim 1 to an oil-producing organism;

culturing said organism; and

isolating the oils or triglycerides.

10. The process of claim 9, further comprising liberating said oils or triglycerides from said organism.

11. The process of claim 9, further comprising introducing at least one further nucleic acid to the organism, wherein said at least one further nucleic acid encodes a polypeptide with a desaturase activity, which is functional or non-functional, and said at least one further nucleic acid contains a sequence selected from the group consisting of:

a sequence that encodes a Δ

-5-desaturase, a Δ

-6-desaturase, a Δ

-8-desaturase or Δ

-12-desaturase;

the sequence of SEQ ID NO;

5;

a sequence derived from the amino acid sequence of SEQ ID NO;

6 by back translation of said amino acid sequence, owing to the degeneracy of the genetic code; and

a derivative of the sequence of SEQ ID NO;

5 wherein said derivative encodes a peptide that has at least 90% identity with the amino acid sequences of SEQ ID NO;

6 and the desaturase activity of said peptide is not substantially reduced as compared to the desaturase activity of said polypeptide.

12. The process of claim 9, wherein the unsaturated fatty acids have an increased content of punicic acid.

13. The process of claim 9, wherein the unsaturated fatty acids have an increased content of octadecaconjudienoic fatty acids.

14. The process of claim 9, wherein the unsaturated fatty acids have an increased content of octadecaconjutetraenoic fatty acids.

15. The nucleic acid of claim 1, wherein the polypeptide has a functional desaturase activity.

16. The process of claim 11, wherein the polypeptide has a functional desaturase activity.

17. The process of claim 9, wherein the organism is a plant or a microorganism.

18. A composition comprising oils or triglycerides with an increased content of unsaturated fatty acids prepared by the process of claim 9.

19. A composition comprising oils or triglycerides with an increased content of unsaturated fatty acids prepared by the process of claim 11.

20. A process for generating transgenic plants comprising introducing the construct of claim 4 to a plant.

21. A process for isolating a genomic sequence via homology screening comprising screening a genome with the nucleic acid of claim 1 or a fragment thereof.

22. A process for production of foods, animal feed, cosmetics or pharmaceuticals comprising introducing the nucleic acid of claim 1 to an organism.

31. The process of claim 11, wherein the nucleic acid encodes a non-functional polypeptide.

32. The process of claim 31, wherein the nucleic acid is an antisense nucleic acid.

23. An isolated nucleic acid sequence that encodes a protein that converts a fatty acid of structure I

24. An isolated nucleic acid that encodes a polypeptide with a desaturase activity, wherein said nucleic acid contains a sequence selected from the group consisting of:

the sequence of SEQ ID NO;

5;

a sequence derived from the amino acid sequence of SEQ ID NO;

6 by back translation of said amino acid sequence, owing to the degeneracy of the genetic code;

a derivative of the sequence of SEQ ID NO;

5 that encodes a peptide which has at least 90% identity at the amino acid level with the amino acid sequence of SEQ ID NO;

6, wherein the desaturase activity of said peptide is not substantially reduced as compared with the desaturase activity of said polypeptide.

25. An amino acid sequence encoded by the nucleic acid of claim 24.

26. A nucleic acid construct comprising the nucleic acid of claim 24, linked to one or more regulatory signal sequences.

27. A vector comprising the nucleic acid construct of claim 26.

28. An organism comprising the vector of claim 27.

29. A transgenic plant comprising the nucleic acid of claim 24.

30. A process for isolating a genomic sequence via homology screening comprising screening a genome with the nucleic acid of claim 24 or a fragment thereof.

33. An isolated nucleic acid that encodes a polypeptide with a non-functional desaturase activity, wherein said nucleic acid contains a sequence selected from the group consisting of:

the sequence of SEQ ID NO;

2;

the sequence of SEQ ID NO;

7;

a sequence derived from the amino acid sequence of SEQ ID NO;

3 or SEQ ID NO;

8 by back translation of the respective amino acid sequences, owing to the degeneracy of the genetic code; and

a derivative of the sequence of SEQ ID NO;

2 or SEQ ID NO;

7 wherein said derivative encodes a peptide that has at least 75 % identity at the amino acid level to said polypeptide.

34. The nucleic acid of claim 33, which is an antisense nucleic acid.

35. A nucleic acid construct comprising the nucleic acid of claim 33 linked to one or more regulatory signal sequences.

36. A vector comprising the nucleic acid construct of claim 35.

37. An organism comprising the vector of claim 36.

38. The organism of claim 37, which is a plant, a microorganism or an animal.

39. A transgenic plant comprising the nucleic acid of claim 33.

40. A process for production of oils or triglycerides with an increased content of unsaturated fatty acids, which comprises:

introducing the nucleic acid of claim 33 to an oil-producing organism;

culturing said organism; and

isolating the oils or triglycerides.

41. The process of claim 40, further comprising liberating said oils or triglycerides from said organism.

42. The process of claim 40, further comprising introducing at least one further nucleic acid to the organism, wherein said at least one further nucleic acid encodes a polypeptide with functional or non-functional desaturase activity and said at least one further nucleic acid contains a sequence selected from the group consisting of:

a sequence that encodes a Δ

-5desaturase, a Δ

-6-desaturase, a Δ

-8-desaturase or Δ

-12-desaturase;

the sequence of SEQ ID NO;

5;

a sequence derived from the amino acid sequence of SEQ ID NO;

6 by back translation of said amino acid sequence, owing to the degeneracy of the genetic code; and

a derivative of the sequence of SEQ ID NO;

5 wherein said derivative encodes a peptide that has at least 90% identity with the amino acid sequences of SEQ ID NO;

6.

43. The process of claim 40, wherein the oils or triglycerides have an increased content of punicic acid.

44. The process of claim 40, wherein the oils or triglycerides have an increased content of octadecaconjudienoic fatty acids.

45. The process of claim 40, wherein the oils or triglycerides have an increased content of octadecaconjutetraenoic fatty acids.

51. A process for isolating a genomic sequence via homology screening comprising screening a genome with the nucleic acid of claim 33 or a fragment thereof.

52. A process for production of foods, animal feed, cosmetics or pharmaceuticals comprising introducing the nucleic acid of claim 33 to an organism.

46. A process for production of oils or triglycerides with an increased content of saturated fatty acids, which comprises:

introducing a nucleic acid to an oil-producing organism wherein said nucleic acid encodes a polypeptide with a non-functional desaturase activity, wherein said nucleic acid contains a sequence selected from the group consisting of;

the sequence of SEQ ID NO;

2;

the sequence of SEQ ID NO;

7;

a sequence derived from the amino acid sequence of SEQ ID NO;

3 or SEQ ID NO;

8 by back translation of the respective amino acid sequences, owing to the degeneracy of the genetic code; and

a derivative of the sequence of SEQ ID NO;

2 or SEQ ID NO;

7

wherein said derivative encodes a peptide that has at least 75 % identity at the amino acid level to said polypeptide;

culturing said organism; and

isolating said oils or triglycerides.

47. The process of claim 46, wherein the nucleic acid is an antisense nucleic acid.

48. The process of claim 46, further comprising liberating said oils or triglycerides.

49. The process of claim 46, wherein the oil-producing organism is a plant or a microorganism.

50. A composition comprising oils or triglycerides with an increased content of unsaturated fatty acids prepared by the process of claim 46.

53. An isolated nucleic acid that encodes a polypeptide with a non-functional desaturase activity, wherein said nucleic acid contains a sequence selected from the group consisting of:

the sequence of SEQ ID NO;

5;

a sequence derived from the amino acid sequence of SEQ ID NO;

6 by back translation of said amino acid sequence, owing to the degeneracy of the genetic code; and

a derivative of the sequence of SEQ ID NO;

5 that encodes a peptide which has at least 90% identity at the amino acid level with the amino acid sequence of SEQ ID NO;

6.

54. The nucleic acid of claim 53, which is an antisense nucleic acid.

55. A nucleic acid construct comprising the nucleic acid sequence of claim 53, which is linked to one or more regulatory signal sequences.

56. A vector comprising the nucleic acid construct of claim 55.

57. An organism comprising the vector of claim 56.

58. A transgenic plant comprising the nucleic acid of claim 53.