Nucleic Acids Encoding Desaturases and Modified Plant Oil

• US 20110162105A1
• Filed: 08/25/2009
• Published: 06/30/2011
• Est. Priority Date: 08/26/2008
• Status: Active Grant

• Current Assignee: BASF Plant Science GmbH (BASF SE), Bioriginal Food & Science Corp. (Omega Protein Corporation)
• Original Assignee: BASF Plant Science GmbH (BASF SE), Bioriginal Food & Science Corp. (Omega Protein Corporation)
• Inventors: Senger, Toralf, Qiu, Xiao, Wu, Guohai, Zank, Thorsten, Bauer, Jörg
• Granted Patent: US 9,090,902 B2
• Time in Patent Office: Days
• US Class Current: 800/281
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 5/14 Plant cells
• CPC Class Codes: C12N 9/00 Enzymes; Proenzymes; Compositions thereof preparations containing enzymes for cleaning teeth A61K8/66, A61Q11/00; medicinal preparations containing enzymes or proenzymes A61K38/43; enzyme containing detergent compositions C11D; enzymes with nucleic acid structure, e.g. ribozymes, C12N15/113; Processes for preparing, activating, inhibiting, separating or purifying enzymes preparation of malt C12C1/00
• CPC Class Codes: C12N 9/0004 Oxidoreductases (1.)

Abstract

The present invention relates to nucleic acids derived from Drechslera tritici-repentis, Cylindorcarpon herteronema, Diploida natalensis, Stagonospora nodorum, Microdochium nivalae and Periplaneta americana. The invention also relates to the individual coding sequences and to proteins encoded by these sequences in combination with other sequences as well as to a process for converting oleic acid to linoleic acid to Iinoleic acid and the production of arachidonic acid, eicosapentaenoic acid and/or docosahexaenoic acid in a plant.

Claims

1. A polynucleotide comprising a nucleic acid selected from the group consisting of:

a) a nucleic acid having a nucleic acid sequence as shown in any one of SEQ ID NO;

1, 3, 5, 7, 31, 33, 35 or 37;

b) a nucleic acid encoding a polypeptide having an amino acid sequence as shown in any one of SEQ ID NO;

2, 4, 6, 8, 32, 34, 36, or 38;

c) a nucleic acid which has a nucleic acid sequence being at least 60% identical to the nucleic acid sequence as shown in any one of SEQ ID NO;

1, 3, 5, 7, 31, 33, 35 or 37, wherein said nucleic acid encodes a polypeptide having desaturase activity;

d) a nucleic acid encoding a polypeptide having an amino acid sequence being at least 74.1% identical to the amino acid sequence shown in any one of SEQ ID NO;

2, 4, 6, 8, 32, 34, 36, or 38, wherein said nucleic acid encodes a polypeptide having desaturase activity;

e) a nucleic acid which hybridizes under stringent hybridization conditions to the nucleic acid of any one of a) to d), wherein said nucleic acid encodes a polypeptide having desaturase activity;

f) a nucleic acid encoding a fragment of a polypeptide encoded by the nucleic acid sequence of any one of a) to e) having desaturase activity; and

g) a nucleic acid comprising at least 15 contiguous nucleotides of the nucleic acid of any one of a) to f).

2. A vector comprising the polynucleotide of claim 1.

3. The vector of claim 2, which is an expression vector.

4. The vector of claim 3, wherein the said polynucleotide is under the control of a seed-specific promoter.

5. The vector of claim 4, wherein the seed-specific promoter is selected from the group consisting of Conlinin 1, Conlinin 2, napin, USP, LeB4, Arc, Fae, ACP, LuPXR, SBP and LuFad3.

6. A host cell comprising the polynucleotide of claim 1 or transformed with an expression vector comprising said polynucleotide.

7. The host cell of claim 6, wherein said host cell is a plant cell.

8. The host cell of claim 7, wherein said plant cell is a cell obtained from an oilseed crop.

9. The host cell of claim 8, wherein the oilseed crop is selected from the group consisting of flax (Linum sp.), rapeseed (Brassica sp.), soybean (Glycine and Soja sp.), sunflower (Helianthus sp.), cotton (Gossypium sp.), corn (Zea mays), olive (Olea sp.), safflower (Carthamus sp.), cocoa (Theobroma cacoa), peanut (Arachis sp.), hemp, camelina, crambe, oil palm, coconuts, groundnuts, sesame seed, castor bean, lesquerella, tallow tree, sheanuts, tungnuts, kapok fruit, poppy seed, jojoba seeds and perilla.

10. The host cell of claim 6, wherein said cell is a microbial cell.

11. The host cell of claim 10, wherein the microbial cell is selected from the group consisting of Candida, Cryptococcus, Lipomyces, Rhodosporidium, Yarrowia, Thraustochytrium, Pythium, Schizochytriwn and cythecodinium.

12. A plant or plant seed comprising the polynucleotide of claim 1, a vector comprising said polynucleotide, or a host cell comprising said polynucleotide or transformed with said vector.

13. A method of producing a polypeptide comprising culturing the host cell of claim 6 in an appropriate culture medium to, thereby, produce the polypeptide encoded by said polynucleotide.

14. A polypeptide encoded by the polynucleotide of claim 1.

15. A method for manufacturing an unsaturated fatty acid, comprising culturing the host cell of claim 6 or a plant or plant seed comprising said host cell such that the unsaturated fatty acid is produced.

17. The method of claim 15, wherein said method further comprises a step of recovering the unsaturated fatty acid from said culture.

18. A method of manufacturing an unsaturated fatty acid comprising contacting a composition comprising at least one desaturase target molecule with at least one polypeptide of claim 14 under conditions such that the unsaturated fatty acid is produced.

19. The method of claim 18, wherein the desaturase target molecule is oleic acid, linoleic acid, γ

-linolenic acid, α

-linolenic acid, dihomo-γ

-linolenic acid, stearidonic acid, eicosatetraenoic acid (n-3), arachidonic acid, eicosapentaenoic acid and docosapentaenoic acid.

20. The method of claim 15, wherein the unsaturated fatty acid is selected from the group consisting of ARA 20:

4 (5,8,11,14), EPA 20;

5 (5,8,11,14,17), and DHA 22;

6 (4,7,10,13,16,19).

21. A method for the manufacture of oil comprising the steps of the method of claim 15 and a further step of formulating or isolating an oil comprising said unsaturated fatty acid.

22. A method of producing a host cell, plant or plant seed capable of generating an unsaturated fatty acid comprising introducing into said host cell, plant or plant seed the polynucleotide of claim 1 or a vector comprising said polynucleotide.

23. An oil produced by the plant or plant seed of claim 12.

26. A method of modulating the production of an unsaturated fatty acid, comprising culturing the host cell of claim 6 or a plant or plant seed comprising said host cell such that modulation of the production of the unsaturated fatty acid occurs.

27. The method of claim 26, wherein said method further comprises a step of recovering the unsaturated fatty acid from said culture.

28. The method of claim 26, wherein the unsaturated fatty acid is selected from the group consisting of ARA 20:

4 (5,8,11,14), EPA 20;

5 (5,8,11,14,17), and DHA 22;

6 (4,7,10,13,16,19).

29. The method of claim 18, wherein the unsaturated fatty acid is selected from the group consisting of ARA 20:

4 (5,8,11,14), EPA 20;

5 (5,8,11,14,17), and DHA 22;

6 (4,7,10,13,16,19).

30. An oil obtainable by the method of claim 29.

16. (canceled)

24. (canceled)

25. A cell comprising a nucleic acid molecule selected from the group consisting of:

a) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO;

1, 3, 5, 7, 31, 33, 35 or 37, wherein the nucleic acid molecule is disrupted by at least one technique selected from the group consisting of a point mutation, a truncation, an inversion, a deletion, an addition, a substitution and homologous recombination;

b) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO;

1, 3, 5, 7, 31, 33, 35 or 37, wherein the nucleic acid molecule comprises one or more nucleic acid modifications as compared to the sequence set forth in SEQ ID NO;

1, 3, 5, 7 or 9, wherein the modification is selected from the group consisting of a point mutation, a truncation, an inversion, a deletion, an addition and a substitution; and

c) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO;

1, 3, 5, 7, 31, 33, 35 or 37, wherein the regulatory region of the nucleic acid molecule is modified relative to the wild-type regulatory region of the molecule by at least one technique selected from the group consisting of a point mutation, a truncation, an inversion, a deletion, an addition, a substitution and homologous recombination.