Elongase gene and method for producing polyunsaturated fatty acids

• US 7,179,647 B2
• Filed: 01/13/2003
• Issued: 02/20/2007
• Est. Priority Date: 01/30/2002
• Status: Expired due to Term

• Current Assignee: BASF Plant Science GmbH (BASF SE)
• Original Assignee: BASF Plant Science GmbH (BASF SE)
• Inventors: Lerchl, Jens, Zank, Thorsten, Heinz, Ernst
• Primary Examiner(s): Prouty; Rebecca E.
• Assistant Examiner(s): MEAH, MOHAMMAD Y
• Application Number:
  US10/502,083
• Publication Number:
  US 20050214761A1
• Time in Patent Office: 1,499 Days
• Field of Search: 435/183, 435/69.1, 800/295
• US Class Current: 435/325
• 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 7/6427 Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
• 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 triacyiglycerol 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 C₁₈-fatty acids with at least two double bonds in the fatty acid by at least two carbon atoms, wherein 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

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

2; and

c) derivatives of the sequence of SEQ ID NO;

1 which encode polypeptides with at least 95% homology with the amino acid sequence of SEQ ID NO;

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

2. The nucleic acid sequence of claim 1, wherein the sequence is derived from an Oomycete.

3. The nucleic acid sequence of claim 1, wherein the sequence is derived from Phytophthora.

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

5. The gene construct of claim 4, wherein the one or more regulatory signals enhance gene expression.

6. A vector comprising the gene construct of claim 4.

10. 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.

11. 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.

12. 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.

13. The nucleic acid of claim 12, wherein the preference is at least a factor of 1.5.

7. An isolated host cell 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 and 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;

2; and

c) derivatives of the sequence of SEQ ID NO;

1 which encode polypeptides with at least 95% homology with the amino acid sequence of SEQ ID NO;

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

8. The isolated host cell of claim 7, wherein the host cell is a microorganism, a nonhuman animal or a plant cell.

9. The isolated host cell of claim 7, wherein the host cell is a transgenic plant cell.