Marchantiales-Derived Unsaturated Fatty Acid Synthetase Genes And Use Of The Same

• US 20080057495A1
• Filed: 12/22/2004
• Published: 03/06/2008
• Est. Priority Date: 12/22/2003
• Status: Active Grant

• Current Assignee: Suntory Holdings Limited (Kotobuki Realty Co., Ltd.)
• Original Assignee: Suntory Ltd (Kotobuki Realty Co., Ltd.)
• Inventors: Ohyama, Kanji
• Granted Patent: US 7,915,487 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• 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: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12N 9/93 Ligases (6)

Abstract

A Δ5 fatty acid desaturase gene, a Δ6 fatty acid desaturase gene, and a Δ6 fatty-acid-chain elongase gene are isolated from a single species of Marchantiales. By introducing these genes into higher plants, transformed plants which can produce arachidonic acid and eicosapentaenoic acid (EPA) are obtained.

Claims

1. An isolated Marchantiales-derived gene that hybridizes under stringent conditions with all of or part of SEQ ID NO:

1 or a complementary sequence thereof, and wherein said gene encodes a protein having Δ

6 fatty acid desaturating activity.

2. The isolated gene of claim 1, wherein said gene (a) consists of a nucleotide sequence of SEQ ID NO:

1, or (b) hybridizes under stringent conditions to SEQ ID NO;

1 or a complementary sequence thereto.

3. The isolated gene of claim 1, wherein said gene (a) comprises a nucleotide sequence of nucleotides 253 to 1698 of SEQ ID NO:

1, or (b) hybridizes under stringent conditions to a nucleotide sequence comprising nucleotides 253 to 1698 of SEQ ID NO;

1 or its or a complementary nucleotide sequence thereto.

13. A protein encoded by a gene of claim 1.

17. An isolated antibody which recognizes a protein of claim 13.

18. A recombinant expression vector which comprises a gene of claim 1.

19. A transformant comprising a gene of claim 1.

20. A plant into which a gene of claim 1 is expressed, a progeny of the plant, a vegetatively propagated plant having the same characteristics, or a tissue of the plant.

21. A plant of claim 20, wherein said plant has a modified fatty acid composition a progeny of the plant, a vegetatively propagated plant having the same characteristics, or a tissue of the plant.

22. A reproductive material of a plant of claim 20.

23. A method of producing fatty acids comprising growing a plant of claim 20 and obtaining said fatty acids from said plant or plant tissue.

24. A composition comprising at least one compound selected from the group consisting of:

γ

-linolenic acid;

dihomo-γ

-linolenic acid;

arachidonic acid;

stearidonic acid;

eicosatetraenoic acid; and

eicosapentaenoic acids, wherein said compound is obtained by a method of claim 23.

25. A method of modifying a fatty acid composition comprising expressing a gene of claim 1.

26. A gene detecting instrument comprising as a probe at least a portion of a nucleotide sequence, or its complementary sequence, of a gene of claim 1.

27. A screening method of a gene or substance that regulates a protein of SEQ ID NO:

2, using a protein of claim 13.

28. A gene or substance obtained by a screening method of claim 27.

4. An isolated gene encoding a Marchantiales-derived protein having Δ

6 fatty acid desaturating activity, wherein said protein has an amino acid sequence of SEQ ID NO;

2, or wherein said gene encodes a protein with an amino acid sequence that has been modified by substitution, deletion, insertion, and/or addition of one or more amino acids of SEQ ID NO;

2.

5. An isolated Marchantiales-derived gene that hybridizes under stringent conditions to all of or part of SEQ ID NO:

3 or a complementary sequence thereof, and wherein said gene encodes a protein having Δ

6 chain elongating activity.

6. The isolated gene of claim 5, wherein said gene (a) consists of a nucleotide sequence of SEQ ID NO:

3, or (b) hybridizes under stringent conditions to SEQ ID NO;

3 or a complementary sequence thereto.

7. The isolated gene of claim 5, wherein said gene (a) comprises a nucleotide sequence of nucleotides 194 to 1066 of SEQ ID NO:

3, or (b) hybridizes under stringent conditions to a nucleotide sequence of nucleotides 194 to 1066 of SEQ ID NO;

1 or a complementary nucleotide sequence thereto.

8. An isolated gene encoding a Marchantiales-derived protein having Δ

6 chain elongating activity, wherein said protein has an amino acid sequence of SEQ ID NO;

4, or wherein said gene encodes a protein with an amino acid sequence that has been modified by substitution, deletion, insertion, and/or addition of one or more amino acids of SEQ ID NO;

4.

9. An isolated Marchantiales-derived gene that hybridizes under stringent conditions to all of or part of SEQ ID NO:

5 or a complementary sequence thereto, wherein said gene, encodes a protein having Δ

5 fatty acid desaturating activity.

10. The isolated gene of claim 9, wherein said gene (a) consists of a nucleotide sequence of SEQ ID NO:

5, or (b) hybridizes under stringent conditions to SEQ ID NO;

5 or a complementary sequence thereto.

11. The isolated gene of claim 9, wherein said gene (a) comprises a nucleotide sequence of nucleotides 375 to 1829 of SEQ ID NO:

5, or (b) hybridizes under stringent conditions to a nucleotide sequence of nucleotides 375 to 1829, of SEQ ID NO;

5 or a complementary nucleotide sequence thereto.

12. An isolated gene encoding a Marchantiales-derived protein having Δ

5 fatty acid desaturating activity, wherein said protein has an amino acid sequence of SEQ ID NO;

6, or wherein said gene encodes a protein with an amino acid sequence that has been modified by substitution, deletion, insertion, and/or addition of one or more amino acids of SEQ ID NO;

6.

14. An isolated protein (a) comprising SEQ ID NO:

2, or (b) consisting of an amino acid sequence that has been modified by substitution, deletion, insertion, and/or addition of one or more amino acids of SEQ ID NO;

2, and wherein said protein has Δ

6 fatty acid desaturating activity.

15. An isolated protein (a) comprising SEQ ID NO:

4, or (b) consisting of an amino acid sequence that has been modified by substitution, deletion, insertion, and/or addition of one or more amino acids of SEQ ID NO;

4, and wherein said protein has Δ

6 chain elongating activity.

16. An isolated protein (a) comprising SEQ ID NO:

6, or (b) consisting of an amino acid sequence that has been modified by substitution, deletion, insertion, and/or addition of one or more amino acids of SEQ ID NO;

6, and wherein said protein has Δ

5 fatty acid desaturating activity.