Method for producing unsaturated ω-3-fatty acids in transgenic organisms

• US 7,777,098 B2
• Filed: 02/23/2005
• Issued: 08/17/2010
• Est. Priority Date: 02/27/2004
• Status: Active Grant

• Current Assignee: BASF Plant Science GmbH (BASF SE)
• Original Assignee: BASF Plant Science GmbH (BASF SE)
• Inventors: Cirpus, Petra, Zank, Thorsten, Bauer, Jörg, Heinz, Ernst
• Primary Examiner(s): McElwain; Elizabeth F
• Application Number:
  US10/590,958
• Publication Number:
  US 20070224661A1
• Time in Patent Office: 2,001 Days
• Field of Search: None
• US Class Current: 800/281
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/0004 Oxidoreductases (1.)
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)

Abstract

The present invention relates to a process for production of unsaturated ω-3-fatty acids and to a process for production of triglycerides with an elevated content of unsaturated fatty acids, especially of ω-3-fatty acids having more than three double bonds. The invention relates to the production of a transgenic organism, preferably of a transgenic plant or of a transgenic microorganism, with an elevated content of unsaturated ω-3-fatty acids, oils or lipids having ω-3-double bonds as the result of the expression of an ω-3-desaturase from fungi of the family Pythiaceae such as the genus Phytophtora, for example of the genus and species Phytophtora infestans.

Claims

1. A process for production of compounds comprising one or more C18-, C20-, and/or C22-polyunsaturated fatty acids in a transgenic organism comprising:

a) introducing, into an organism, at least one nucleic acid sequence which encodes an ω

-3-desaturase that is capable of desaturating C20- or C22-fatty acids, and

b) culturing the organism under conditions which permits the production of one or more C18-, C20-, and/or C22-polyunsaturated fatty acids,

wherein the ω

-3-desaturase is encoded by a nucleic acid sequence comprising;

i) the nucleic acid sequence of SEQ ID NO;

1,

ii) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO;

2,

iii) a nucleic acid sequence having at least 95% identity with the nucleic acid sequence of SEQ ID NO;

1, or

iv) a nucleic acid sequence encoding an amino acid sequence having at least 95% identity with the amino acid sequence of SEQ ID NO;

2.

2. The process according to claim 1, further comprising introducing into the organism at least one nucleic acid sequence coding for a polypeptide with Δ

9-elongase, Δ

6-desaturase, Δ

8-desaturase, Δ

6-elongase, Δ

5-desaturase, Δ

5-elongase or Δ

4-desaturase activity.

3. The process according to claim 1, wherein the one or more C18-, C20-, and/or C22-polyunsaturated fatty acids have at least two double bonds.

4. The process according to claim 1, wherein the transgenic organism is a transgenic microorganism or a transgenic plant.

5. The process according to claim 1, wherein the transgenic organism is an oil-producing plant, a vegetable producing plant, or an ornamental plant.

6. The process according to claim 1, wherein the transgenic organism is a transgenic plant selected from the group of the plant families consisting of Adelotheciaceae, Anacardiaceae, Asteraceae, Apiaceae, Betulaceae, Boraginaceae, Brassicaceae, Bromeliaceae, Caricaceae, Cannabaceae, Convolvulaceae, Chenopodiaceae, Crypthecodiniaceae, Cucurbitaceae, Ditrichaceae, Elaeagnaceae, Ericaceae, Euphorbiaceae, Fabaceae, Geraniaceae, Gramineae, Juglandaceae, Lauraceae, Leguminosae, Linaceae and Prasinophyceae.

7. The process according to claim 1, wherein the one or more C18-, C20-, and/or C22-polyunsaturated fatty acids are isolated from the organism in the form of their oils, lipids or free fatty acids.

8. The process according to claim 1, wherein the one or more C18-, C20-, and/or C22-polyunsaturated fatty acids are isolated in a concentration of at least 5% by weight based on the total lipid content of the transgenic organism.

9. The process according to claim 1, wherein the ω

-3-desaturase is capable of desaturating C20-fatty acids.

10. The process according to claim 1, wherein the ω

-3-desaturase is capable of desaturating C18-, C20- and C22-fatty acids.

11. A process for increasing the content of C18-, C20- and/or C22-polyunsaturated fatty acids in oils, lipids, or fatty acids in an organism comprising:

a) introducing, into an organism, at least one nucleic acid sequence which encodes an ω

-3-desaturase that is capable of desaturating C20- or C22-fatty acids, and

b) culturing the organism under conditions which permits the production of C18-, C20- and/or C22-polyunsaturated fatty acids

wherein the ω

-3-desaturase is encoded by a nucleic acid sequence comprising;

i) the nucleic acid sequence of SEQ ID NO;

1,

ii) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO;

2,

iii) a nucleic acid sequence having at least 95% identity with the nucleic acid sequence of SEQ ID NO;

1, or

iv) a nucleic acid sequence encoding an amino acid sequence having at least 95% identity with the amino acid sequence of SEQ ID NO;

2.

12. The process according to claim 11, further comprising introducing into the organism at least one nucleic acid sequence coding for a polypeptide with Δ

9-elongase, Δ

6-desaturase, Δ

8-desaturase, Δ

6-elongase, Δ

5-desaturase, Δ

5-elongase or Δ

4-desaturase activity.

13. The process according to claim 11, wherein the C18-, C20- and/or C22-polyunsaturated fatty acids have at least two double bonds.

14. The process according to claim 11, wherein the transgenic organism is a transgenic microorganism or a transgenic plant.

15. The process according to claim 11, wherein the transgenic organism is an oil-producing plant, a vegetable producing plant, or an ornamental plant.

16. The process according to claim 11, wherein the transgenic organism is a transgenic plant selected from the group of the plant families consisting of Adelotheciaceae, Anacardiaceae, Asteraceae, Apiaceae, Betulaceae, Boraginaceae, Brassicaceae, Bromeliaceae, Caricaceae, Cannabaceae, Convolvulaceae, Chenopodiaceae, Crypthecodiniaceae, Cucurbitaceae, Ditrichaceae, Elaeagnaceae, Ericaceae, Euphorbiaceae, Fabaceae, Geraniaceae, Gramineae, Juglandaceae, Lauraceae, Leguminosae, Linaceae and Prasinophyceae.

17. The process according to claim 11, further comprising isolating the oils, lipids, or fatty acids from the organism.

18. The process according to claim 11, wherein the ω

-3-desaturase is capable of desaturating C20-fatty acids.

19. The process according to claim 11, wherein the ω

-3-desaturase is capable of desaturating C18-, C20- and C22-fatty acids.

20. A process for production of compounds comprising one or more C18-, C20-, and/or C22-polyunsaturated fatty acids in a transgenic organism comprising:

a) introducing into an organism, at least one nucleic acid sequence which encodes an ω

-3-desaturase that is capable of desaturating C22;

4 ω

-6-fatty acid to C22;

5 ω

-3-fatty acid, and

b) culturing the organism under conditions which permits the production of one or more C18-, C20-, and/or C22-polyunsaturated fatty acids.

21. The process according to claim 20, wherein the one or more C18-, C20-, and/or C22-polyunsaturated fatty acids have at least two double bonds.

22. The process according to claim 20, wherein the transgenic organism is a transgenic microorganism or a transgenic plant.