Method of producing polyunsaturated fatty acids, novel biosynthesis genes, and novel plant expression constructs

• US 7,615,679 B2
• Filed: 01/18/2002
• Issued: 11/10/2009
• Est. Priority Date: 01/19/2001
• Status: Expired due to Fees

• Current Assignee: BASF Plant Science GmbH (BASF SE)
• Original Assignee: BASF Plant Science GmbH (BASF SE)
• Inventors: Domergue, Frederic, Lerchl, Jens, Renz, Andreas, Heinz, Ernst, Zahringer, Ulrich
• Primary Examiner(s): McElwain; Elizabeth F
• Application Number:
  US10/250,553
• Publication Number:
  US 20040053379A1
• Time in Patent Office: 2,853 Days
• Field of Search: None
• US Class Current: 800/281
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• 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: C12P 7/6427 Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
• CPC Class Codes: C12P 7/6432 Eicosapentaenoic acids [EPA]
• CPC Class Codes: C12P 7/6434 Docosahexenoic acids [DHA]
• 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 present invention relates to a method for the production of unsaturated fatty acids with at least two double bonds. The invention further relates to the use of the nucleic acid sequence of SEQ ID NO: 1 encoding a polypeptide having delta-5 desaturase activity in the method and for generating a transgenic organism, preferably a transgenic plant or microorganism with an increased content of fatty acids, oil or lipids with unsaturated long chain fatty acids, to their homologs or derivatives, and to gene constructs encompassing these genes. The invention also relates to the use of the nucleic acid sequences alone or in combination with further desaturases and/or elongase genes in the method and for generating transgenic organisms.

Claims

1. A method for the production of fatty acid esters with an increased content of polyunsaturated fatty acids with at least two double bonds, which comprises introducing, into a fatty-acid-ester-producing organism, at least one nucleic acid sequence selected from the group consisting of

a) a nucleic acid sequence with the sequence shown in SEQ ID NO:

1,

b) a nucleic acid sequence encoding the amino acid sequence shown in SEQ ID NO;

2,

c) a derivative of the nucleic acid sequence shown in SEQ ID NO;

1, which encodes a polypeptide having at least 90% homology with the amino acid sequence shown in SEQ ID NO;

2, and having Δ

5-desaturase activity,

growing the organism, and isolating the fatty acid esters present in the organism, wherein the organism is a microorganism, a yeast, or a plant or plant cell.

2. The method as claimed in claim 1, wherein the fatty acid esters produced by the method comprise polyunsaturated C₁₈-, C₂₀- or C₂₂-fatty acid molecules with at least two double bonds in the fatty acid ester.

3. The method as claimed in claim 1, wherein the C₁₈-, C₂₀- or C₂₂-fatty acid molecules are isolated from the organism in the form of an oil or lipid.

4. The method as claimed in claim 1, wherein a combination of the nucleic acid sequences of SEQ ID NO:

1, SEQ ID NO;

3 and SEQ ID NO;

9 is introduced into the fatty-acid-ester-producing organism.

5. The method as claimed in claim 1, wherein the organism is an oil crop plant or plant cell.

6. The method as claimed in claim 1, wherein the organism is a transgenic plant.

7. The method as claimed in claim 1, wherein the fatty acid esters contain C₁₈-, C₂₀- or C₂₂-fatty acids with three, four or five double bonds in the fatty acid ester.

8. The method as claimed in claim 1, wherein the polyunsaturated fatty acids contained in the fatty acid esters are liberated.

9. An isolated nucleic acid sequence encoding a polypeptide with desaturase activity, selected from the group consisting of:

a) a nucleic acid sequence with the sequence shown in SEQ ID NO;

1,

b) a nucleic acid sequence encoding the amino acid sequence shown in SEQ ID NO;

2,

c) a derivative of the nucleic acid sequence shown in SEQ ID NO;

1, which encodes a polypeptide having at least 90% homology with the amino acid sequence shown in SEQ ID NO;

2, and having Δ

5-desaturase activity.

10. A nucleic acid construct comprising the nucleic acid sequence as claimed in claim 9, wherein the nucleic acid sequence is linked to one or more regulatory signals.

11. A vector comprising the nucleic acid sequence as claimed in claim 9.

13. The isolated nucleic acid sequence of claim 9, wherein the nucleic acid sequence encodes a polypeptide having at least 95% homology with the amino acid sequence shown in SEQ ID NO:

2.

12. An isolated nucleic acid sequence encoding a polypeptide comprising a sequence having at least 90% homology with the amino acid sequence shown in SEQ ID NO:

2.