Δ-4 desaturases from Euglena gracilis, expressing plants, and oils containing PUFA

• US 7,629,503 B2
• Filed: 04/06/2004
• Issued: 12/08/2009
• Est. Priority Date: 04/08/2003
• Status: Expired due to Fees

• Current Assignee: BASF Plant Science GmbH (BASF SE)
• Original Assignee: BASF Plant Science GmbH (BASF SE)
• Inventors: Meyer, Astrid, Cirpus, Petra, Heinz, Ernst, Zahringer, Ulrich, Bauer, Jorg
• Primary Examiner(s): McElwain; Elizabeth F
• Application Number:
  US10/552,127
• Publication Number:
  US 20060218668A1
• Time in Patent Office: 2,072 Days
• Field of Search: None
• US Class Current: 800/298
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)

Abstract

The present invention relates to an improved process for the specific preparation of unsaturated ω-3 fatty acids, and to a process for preparing triglycerides having an increased content of unsaturated fatty acids, particularly of ω-3 fatty acids having more than three double bonds. The invention relates to the preparation of a transgenic organism, preferably a transgenic plant or a transgenic microorganism, having increased content of fatty acids, oils or lipids having Δ-4 double bonds owing to the expression of a Δ-4-desaturase from Euglena gracilis.

The invention additionally relates to expression cassettes comprising a nucleic acid sequence, a vector and organisms comprising at least one nucleic acid sequence or one expression cassette. The invention additionally relates to unsaturated fatty acids and to triglycerides having an increased content of unsaturated fatty acids and to the use thereof.

Fatty acids and triglycerides have a large number of uses in the food industry, in animal nutrition, cosmetics and in the drugs sector. They are suitable for a wide variety of uses depending on whether they are free saturated or unsaturated fatty acids or triglycerides having an increased content of saturated or unsaturated fatty acids.

Claims

1. An isolated nucleic acid comprising a nucleotide sequence selected from the group consisting of:

a) the nucleotide sequence as depicted in SEQ ID NO;

1,

b) a nucleotide sequence which codes for a polypeptide having the amino acid sequence of SEQ ID NO;

2, and

c) a nucleotide sequence which codes for a polypeptide having at least 95% homology at the amino acid level with SEQ ID NO;

2, wherein the polypeptide has Δ

-4-desaturase activity.

2. The isolated nucleic acid of claim 1, wherein the sequence is derived from a plant.

3. The isolated nucleic acid of claim 1, wherein the sequence is derived from the class of Euglenophyceae.

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

5. The gene construct of claim 4, wherein the gene construct comprises additional biosynthesis genes of fatty acid or lipid metabolism selected from the group consisting of acyl-CoA dehydrogenase(s), acyl-ACP [=acyl carrier protein]desaturase(s), acyl-ACP thioesterase(s), fat acid acyltransferase(s), acyl-CoA:

lysophospholipid acyltransferase(s), fatty acid synthase(s), fatty acid hydroxylase(s), acetyl-coenzyme A carboxylase(s), acyl-coenzyme A oxidase(s), fatty acid desaturase(s), fatty acid acetylenases, lipoxygenases, triacylglycerol lipases, allene oxide synthases, hydroperoxide lyases, and fatty acid elongase(s).

6. The gene construct of claim 4, wherein the gene construct comprises additional biosynthesis genes of fatty acid or lipid metabolism selected from the group consisting of Δ

-4-desaturase, Δ

-5-desaturase, Δ

-6-desaturase, Δ

-8-desaturase, Δ

-9-desaturase, Δ

-12-desaturase, Δ

-5-elongase, Δ

-6-elongase, and Δ

-9-elongase.

7. A vector comprising the nucleic acid of claim 1.

8. A transgenic nonhuman organism comprising at least one nucleic acid of claim 1, wherein the nonhuman organism is a microorganism, a yeast, or a plant.

9. The transgenic nonhuman organism of claim 8, wherein the organism is a plant.

10. A process for producing polyunsaturated fatty acids, comprising growing a transgenic organism which comprises the nucleic acid of claim 1, producing polyunsaturated fatty acids in said organism, and recovering the polyunsaturated fatty acids, wherein the organism is a yeast or a plant.

11. The process of claim 10, wherein docosahexaenoic acid is produced in the process.

12. The process of claim 10, wherein the polyunsaturated fatty acids are isolated from the organism in the form of an oil, lipid or a free fatty acid.

13. The process of claim 10, wherein the organism is a transgenic plant.