Novel elongase gene and method for producing polyunsaturated fatty acids

• US 20050214761A1
• Filed: 01/13/2003
• Published: 09/29/2005
• Est. Priority Date: 01/30/2002
• Status: Active Grant

• Current Assignee: BASF Plant Science GmbH (BASF SE)
• Original Assignee: BASF Plant Science GmbH (BASF SE)
• Inventors: Lerchl, Jens, Zank, Thorsten, Heinz, Ernst
• Granted Patent: US 7,179,647 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• 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 invention relates to a novel elongase gene with the sequence SEQ ID NO:1 or its homologs, derivatives or analogs, to a gene construct comprising this gene or its homologs, derivatives and analogs, and to its use. The invention also relates to vectors or organisms comprising an elongase gene with the sequence SEQ ID NO:1 or its homologs, derivatives or analogs. Furthermore, the invention relates to a process for the preparation of polyunsaturated fatty acids and to a process for introducing DNA into organisms which produce large amounts of oils and, in particular, oils with a high content of unsaturated fatty acids. Moreover, the invention relates to an oil and/or a fatty acid preparation with a higher content of polyunsaturated fatty acids with at least two double bonds and/or a triacylglycerol preparation with a higher content of polyunsaturated fatty acids with at least two double bonds.

Claims

1. An isolated nucleic acid encoding a polypeptide that elongates C₁₆- or C₁₈fatty acids with at least two double bonds in the fatty acid by at least two carbon atoms, wherein one or more of C_18.3^Δ

5t,9,12, C<sub>20;

3</sub><sup>Δ

8,11,14</sup>, C<sub>20;

4</sub><sup>Δ

5,8,11,14</sup> and C<sub>20;

50</sub><sup>Δ

5,8,11,14,17</sup> are not elongated.

5. An amino acid sequence encoded within the nucleic acid of claim 1.

6. A gene construct comprising the nucleic acid of claim 1, wherein the nucleic acid is functionally linked to one or more regulatory signals.

7. The gene construct of claim 6, wherein the one or more regulatory signals enhance gene expression.

8. A vector comprising the gene construct of claim 6.

22. The nucleic acid of claim 1, which encodes a polypeptide that elongates C₁₆- or C₁₈-fatty acids with at least three double bonds in the fatty acid.

23. The nucleic acid of claim 1, which encodes a polypeptide that elongates C₁₆- or C₁₈-fatty acids with at least four double bonds in the fatty acid.

24. The nucleic acid of claim 1, wherein the polypeptide shows a preference for elongating C_18:

3<sup>Δ

6,9,12</sup>, C<sub>18;

4</sub><sup>Δ

6,9,12,15</sup>, or C<sub>16;

3</sub><sup>Δ

7,10,13</sup>-fatty acids as compared to one or more of C<sub>18;

2</sub><sup>Δ

9,12</sup>, C<sub>18;

3</sub><sup>Δ

4,7,10</sup>, C<sub>18;

3</sub><sup>Δ

5,8,11</sup>, C<sub>18;

3</sub>^7,10,13, C<sub>18;

3</sub><sup>Δ

8,11,14</sup>, C<sub>18;

3</sub><sup>Δ

9,12,15</sup> or C<sub>18;

3</sub><sup>Δ

5,c9,12</sup>-fatty acids.

25. The nucleic acid of claim 24, wherein the preference is at least a factor of 1.5.

2. An isolated nucleic acid comprising a nucleotide sequence encoding a polypeptide that elongates C₁₆- or C₁₈-fatty acids with at least two double bonds in the fatty acid molecule, wherein said nucleotide sequence is selected from the group consisting of:

a) the sequence of SEQ ID NO;

1;

b) a nucleic acid sequence which, in accordance with degeneracy of the genetic code, is derived from the amino acid sequence of SEQ ID NO;

2; and

c) derivatives of the sequence of SEQ ID NO;

1 which encode polypeptides with at least 50% homology with the amino acid sequences of SEQ ID NO;

2, wherein the sequence acts as a C₁₆- or C₁₈-elongase.

3. The nucleic acid sequence of claim 2, wherein the sequence is derived from an Oomycete.

4. The nucleic acid sequence of claim 2, wherein the sequence is derived from Phytophthora.

9. An organism comprising a recombinant nucleic acid that encodes a polypeptide, which elongates C₁₆- or C₁₈-fatty acids with at least two double bonds in the fatty acid by at least two carbon atoms, wherein one or more of C_18:

3<sup>Δ

5t,9,12</sup>, C<sub>20;

3</sub><sup>Δ

8,11,14</sup>, C<sub>20;

4</sub><sup>Δ

5,8,11,14</sup> and C<sub>20;

5</sub><sup>Δ

5,8,14,17</sup> are not elongated.

10. The organism of claim 9, wherein the organism is a microorganism, a nonhuman animal or a plant.

11. The organism of claim 9, wherein the organism is a transgenic plant.

12. A process for the production of PUFAs, which comprises culturing the organism of claim 9, under conditions in which said PUFAs are formed in the organism.

13. The process of claim 12, wherein the PUFAs prepared by the process are C₂₀- or C₂₂-fatty acid molecules with at least two double bonds in the fatty acid molecule.

14. The process of claim 13, wherein the C₂₀- or C₂₂-fatty acid molecules are isolated from the organism in the form of an oil, a lipid or a free fatty acid.

15. The process of claim 12, wherein the organism is a microorganism, a nonhuman animal or a plant.

16. The process of claim 12, wherein the organism is a transgenic plant.

17. The process of claim 12, wherein the C₁₆- or C₁₈-fatty acid is a fatty acid with three double bonds in the molecule.

18. An oil, lipid or fatty acid or a fraction thereof, prepared by the process of claim 12.

19. An oil, lipid or fatty acid composition which comprises PUFAs and is derived from a transgenic plant.

20. The oil, lipid or fatty acid composition of claim 19, wherein the transgenic plant contains a nucleotide sequence encoding a polypeptide that elongates C₁₆- or C₁₈-fatty acids with at least two double bonds in the fatty acid molecule, wherein said nucleotide sequence is selected from the group consisting of:

the sequence of SEQ ID NO;

1;

a nucleic acid sequence which, in accordance with degeneracy of the genetic code, is derived from the amino acid sequence of SEQ ID NO;

2; and

derivatives of the sequence of SEQ ID NO;

1 which encode polypeptides with at least 50% homology with the sequence encoding the amino acid sequences of SEQ ID NO;

2, the sequence acting as C₁₆- or C₁₈-elongase.

21. Feeding stuffs, foodstuffs, cosmetics or pharmaceuticals comprising the oil, lipid or fatty acid composition of claim 19.