Methods and compositions for synthesis of long chain polyunsaturated fatty acids
First Claim
1. A method for production of γ
- -linolenic acid in a yeast culture, said method comprising;
growing a yeast culture having a plurality of recombinant yeast cells, wherein said yeast cells or an ancestor of said yeast cells were transformed with a vector comprising Mortierella alpina DNA encoding a Δ
6 desaturase polypeptide which converts linoleic acid to γ
-linolenic acid, wherein said DNA is operably associated with an expression control sequence functional in said yeast cells, under conditions whereby said DNA is expressed, whereby γ
-linolenic acid is produced from linoleic acid in said yeast culture.
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Abstract
The present invention relates to fatty acid desaturases able to catalyze the conversion of oleic acid to linoleic acid, linoleic acid to gamma-linolenic acid, or of alpha-linolenic acid to stearidonic acid. Nucleic acid sequences encoding desaturases, nucleic acid sequences which hybridize thereto, DNA constructs comprising a desaturase gene, and recombinant host microorganism or animal expressing increased levels of a desaturase are described. Methods for desaturating a fatty acid and for producing a desaturated fatty acid by expressing increased levels of a desaturase are disclosed. Fatty acids, and oils containing them, which have been desaturated by a desaturase produced by recombinant host microorganisms or animals are provided. Pharmaceutical compositions, infant formulas or dietary supplements containing fatty acids which have been desaturated by a desaturase produced by a recombinant host microorganism or animal also are described.
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Citations
22 Claims
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1. A method for production of γ
- -linolenic acid in a yeast culture, said method comprising;
growing a yeast culture having a plurality of recombinant yeast cells, wherein said yeast cells or an ancestor of said yeast cells were transformed with a vector comprising Mortierella alpina DNA encoding a Δ
6 desaturase polypeptide which converts linoleic acid to γ
-linolenic acid, wherein said DNA is operably associated with an expression control sequence functional in said yeast cells, under conditions whereby said DNA is expressed, whereby γ
-linolenic acid is produced from linoleic acid in said yeast culture. - View Dependent Claims (2, 3, 4, 5)
- -linolenic acid in a yeast culture, said method comprising;
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6. A method for production of stearidonic acid in a yeast culture, said method comprising:
growing a yeast culture having a plurality of recombinant yeast cells, wherein said yeast cells or an ancestor of said yeast cells were transformed with a vector comprising Mortierella alpina DNA encoding a Δ
6 desaturase polypeptide which converts α
-linolenic acid to stearidonic acid, wherein said DNA is operably associated with an expression control sequence functional in said yeast cells, under conditions whereby said DNA is expressed, whereby stearidonic acid is produced from α
-linolenic acid in said yeast culture.- View Dependent Claims (7, 8, 9, 10)
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11. A method for production of linoleic acid in a yeast culture, said method comprising:
growing a yeast culture having a plurality of recombinant yeast cells, wherein said yeast cells or an ancestor of said yeast cells were transformed with a vector comprising Mortierella alpina DNA encoding a Δ
12 desaturase polypeptide which converts oleic acid to linoleic acid, wherein said DNA is operably associated with an expression control sequence functional in said yeast cells, under conditions whereby said DNA is expressed, whereby linoleic acid is produced oleic acid in said yeast culture.- View Dependent Claims (12, 13, 14)
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15. A method for production of stearidonic acid in a eukaryotic cell culture, said method comprising:
growing a eukaryotic cell culture having a plurality of recombinant eukaryotic cells, wherein said recombinant eukaryotic cells or ancestors of said recombinant eukaryotic cells were transformed with a vector comprising Mortierella alpina DNA encoding a polypeptide which converts α
-linolenic to stearidonic acid, wherein said DNA is operably associated with an expression control sequence functional in said recombinant eukaryotic cells, under conditions whereby said DNA is expressed, whereby stearidonic acid is produced from α
-linolenic in said eukaryotic cell culture.- View Dependent Claims (17, 19)
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16. A method for production of linoleic acid in a eukaryotic cell culture, said method comprising:
growing a eukaryotic cell culture having a plurality of recombinant eukaryotic cells, wherein said recombinant eukaryotic cells or ancestors of said recombinant eukaryotic cells were transformed with a vector comprising Mortierella alpina DNA encoding a polypeptide which converts oleic acid to linoleic acid, wherein said DNA is operably associated with an expression control sequence functional in said recombinant eukaryotic cells, under conditions whereby said DNA is expressed, whereby linoleic acid is produced from oleic acid in said eukaryotic cell culture. - View Dependent Claims (18, 20)
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21. A method of making α
- -linolenic acid, said method comprising;
growing a recombinant yeast cell comprising a nucleotide sequence which encodes a functionally active Δ
6 desaturase having the amino acid sequence depicted in SEQ ID NO;
2, and at least one nucleic acid construct comprising a nucleotide sequence which encodes a functionally active Δ
12 desaturase having an amino acid sequence depicted in SEQ ID NO;
4, wherein said nucleic acid constructs are operably associated with transcription control sequences functional in a yeast cell under conditions whereby said nucleotide sequences are expressed, whereby γ
-linolenic acid is produced in said yeast cell. - View Dependent Claims (22)
- -linolenic acid, said method comprising;
Specification