Method of producing polyunsaturated fatty acids, novel biosynthesis genes, and novel plant expression constructs
First Claim
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, SEQ ID NO;
3, SEQ ID NO;
5 or SEQ ID NO;
11, b) nucleic acid sequences which, owing to the degeneracy of the genetic code, are obtained by backtranslating the amino acid sequences shown in SEQ ID NO;
2, SEQ ID NO;
4, SEQ ID NO;
6 or SEQ ID NO;
12, c) derivatives of the nucleic acid sequence shown in SEQ ID NO;
1, SEQ ID NO;
3, SEQ ID NO;
5 or SEQ ID NO;
11, which encode polypeptides with the amino acid sequences shown in SEQ ID NO;
2, SEQ ID NO;
4, SEQ ID NO;
6 or SEQ ID NO;
12 and have at least 50% homology at the amino acid level, without essentially reducing the enzymatic action of the polypeptides, growing the organism, and isolating the fatty acid esters present in the organism.
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Abstract
The present invention relates to a method for the production of unsaturated fatty acids with at least two double bonds and/or a method for the production of triglycerides with an increased content of polyunsaturated fatty acids with at least two double bonds. The invention furthermore relates to the advantageous use of the nucleic acid sequences SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 9, SEQ ID NO: 11 or SEQ ID NO: 18 in the method and for generating a transgenic organism, preferably a transgenic plant or a transgenic microorganism, with an increased content of fatty acids, oils or lipids with unsaturated C18-, C20-, or C22-fatty acids.
The invention furthermore relates to novel desaturases with the [lacuna] in the sequences SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5 and SEQ ID NO: 11 or its homologs, derivatives and analogs, and to gene constructs encompassing these genes or their homologs, derivatives or analogs, and to their use alone or in combination with biosynthesis genes of polyunsaturated fatty acids as shown advantageously in SEQ ID NO: 7 and SEQ ID NO: 9.
In addition, the invention relates to isolated nucleic acid sequences; expression cassettes comprising the nucleic acid sequences, vectors and transgenic organisms comprising at least one nucleic acid sequence or one expression cassette. In addition, the invention relates to unsaturated fatty acids with at least two double bonds and to triglycerides with an increased content of unsaturated fatty acids with at least two double bonds, and to their use.
Moreover, the invention relates to multiexpression cassettes for seed-specific expression, and to vectors or organisms which encompass a desaturase gene alone or in combination with further desaturases with the sequence SEQ ID NO:7 and/or elongase genes with the sequence SEQ ID NO: 9 or its homologs, derivatives or analogs using said expression cassettes.
67 Citations
27 Claims
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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, SEQ ID NO;
3, SEQ ID NO;
5 or SEQ ID NO;
11,b) nucleic acid sequences which, owing to the degeneracy of the genetic code, are obtained by backtranslating the amino acid sequences shown in SEQ ID NO;
2, SEQ ID NO;
4, SEQ ID NO;
6 or SEQ ID NO;
12,c) derivatives of the nucleic acid sequence shown in SEQ ID NO;
1, SEQ ID NO;
3, SEQ ID NO;
5 or SEQ ID NO;
11, which encode polypeptides with the amino acid sequences shown in SEQ ID NO;
2, SEQ ID NO;
4, SEQ ID NO;
6 or SEQ ID NO;
12 and have at least 50% homology at the amino acid level, without essentially reducing the enzymatic action of the polypeptides,growing the organism, and isolating the fatty acid esters present in the organism. - View Dependent Claims (2, 3, 4, 5, 6, 7, 21, 22, 23, 26)
- 1, SEQ ID NO;
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8. An isolated nucleic acid sequence encoding a polypeptide with desaturase activity, selected from the group consisting of:
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a) a nucleic acid sequence with the sequence shown in SEQ ID NO;
1, SEQ ID NO;
3, SEQ ID NO;
5 or SEQ ID NO;
11,b) nucleic acid sequences which, owing to the degeneracy of the genetic code, are obtained by backtranslating the amino acid sequences shown in SEQ ID NO;
2, SEQ ID NO;
4, SEQ ID NO;
6 or SEQ ID NO;
12,c) derivatives of the nucleic acid sequences shown in SEQ ID NO;
1, SEQ ID NO;
3, SEQ ID NO;
5 or SEQ ID NO;
11, which encode polypeptides with the amino acid sequences shown in SEQ ID NO;
2, SEQ ID NO;
4, SEQ ID NO;
6 or SEQ ID NO;
12 and have at least 50% homology at the amino acid level, without essentially reducing the enzymatic action of the polypeptides. - View Dependent Claims (9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 27)
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24. A method of identifying an antagonist or agonist of desaturases, comprising
a) contacting the cells which express the polypeptide of the present invention with a candidate substance; -
b) testing the desaturate activity;
c) comparing the desaturase activity with a standard activity in the absence of the candidate material, where an increase in the desaturase activity beyond the standard indicates that the candidate material is an agonist and a reduction in the desaturase activity indicates that the candidate material is an antagonist.
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Specification