Delta4-desaturase genes and uses thereof
First Claim
1. An isolated nucleotide sequence or fragment thereof comprising or complementary to a nucleotide sequence encoding a polypeptide having desaturase activity, wherein the amino acid sequence of said polypeptide has at least 50% identity to an amino acid sequence selected from the group consisting of SEQ ID NO:
- 18, SEQ ID NO;
19, SEQ ID NO;
20, SEQ ID NO;
21, SEQ ID NO;
37, SEQ ID NO;
46 and SEQ ID NO;
56.
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Accused Products
Abstract
The subject invention relates to the identification of genes involved in the desaturation of polyunsaturated fatty acids at carbon 4 (i.e., “Δ4-desaturase”). In particular, Δ4-desaturase may be utilized, for example, in the conversion of adrenic acid to ω6-docosapentaenoic acid and in the conversion of ω3-docosapentaenoic acid to docosahexaenoic acid. The polyunsaturated fatty acids produced by use of the enzyme may be added to pharmaceutical compositions, nutritional compositions, animal feeds, as well as other products such as cosmetics.
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Citations
31 Claims
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1. An isolated nucleotide sequence or fragment thereof comprising or complementary to a nucleotide sequence encoding a polypeptide having desaturase activity, wherein the amino acid sequence of said polypeptide has at least 50% identity to an amino acid sequence selected from the group consisting of SEQ ID NO:
- 18, SEQ ID NO;
19, SEQ ID NO;
20, SEQ ID NO;
21, SEQ ID NO;
37, SEQ ID NO;
46 and SEQ ID NO;
56. - View Dependent Claims (3, 4, 5, 6)
- 18, SEQ ID NO;
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2. An isolated nucleotide sequence or fragment thereof comprising or complementary to a nucleotide sequence having at least 50% identity to a nucleotide sequence selected from the group consisting of SEQ ID NO:
- 14, SEQ ID NO;
15 and SEQ ID NO;
16, SEQ ID NO;
17, SEQ ID NO;
36, SEQ ID NO;
45 and SEQ ID NO;
55.
- 14, SEQ ID NO;
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7. A purified polypeptide which desaturates polyunsaturated fatty acids at carbon 4 and has an amino acid sequence having at least 50% identity to an amino acid sequence selected from the group consisting of SEQ ID NO:
- 18, SEQ ID NO;
19, SEQ ID NO;
20, SEQ ID NO;
21, SEQ ID NO;
37, SEQ ID NO;
46 and SEQ ID NO;
56.
- 18, SEQ ID NO;
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8. A method of producing a desaturase comprising the steps of:
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a) isolating a nucleotide sequence comprising or complementary to a nucleotide sequence;
i) encoding a polypeptide having an amino acid sequence having at least 50% identity to an amino acid sequence selected from the group consisting of SEQ ID NO;
18, SEQ ID NO;
19, SEQ ID NO;
20, SEQ ID NO;
21, SEQ ID NO;
37, SEQ ID NO;
46 and SEQ ID NO;
56 orii) having at least 50% identity to a nucleotide sequence selected from the group consisting of SEQ ID NO;
14, SEQ ID NO;
15, SEQ ID NO;
16, SEQ ID NO;
17, SEQ ID NO;
36, SEQ ID NO;
45 and SEQ ID NO;
55;
b) constructing a vector comprising;
i) said isolated nucleotide sequence operably linked to ii) a promoter;
c) introducing said vector into a host cell for a time and under conditions sufficient for expression of said desaturase.
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9. A vector comprising:
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a) an isolated nucleotide sequence comprising or complementary to a nucleotide sequence;
i) encoding a polypeptide having an amino acid sequence having at least 50% identity to an amino acid sequence selected from the group consisting of SEQ ID NO;
18, SEQ ID NO;
19, SEQ ID NO;
20, SEQ ID NO;
21, SEQ ID NO;
37, SEQ ID NO;
46 and SEQ ID NO;
56 orii) having at least 50% identity to a nucleotide sequence selected from the group consisting of SEQ ID NO;
14, SEQ ID NO;
15, SEQ ID NO;
16, SEQ ID NO;
17, SEQ ID NO;
36, SEQ ID NO;
45 and SEQ ID NO;
55, operably linked tob) a promoter. - View Dependent Claims (10, 11, 12, 13, 14)
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15. A method for producing a polyunsaturated fatty acid comprising the steps of:
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a) isolating a nucleotide sequence comprising or complementary to a nucleotide sequence;
i) encoding a polypeptide having an amino acid sequence having at least 50% identity to an amino acid sequence selected from the group consisting of SEQ ID NO;
18, SEQ ID NO;
19, SEQ ID NO;
20, SEQ ID NO;
21, SEQ ID NO;
37, SEQ ID NO;
46 and SEQ ID NO;
56 orii) having at least 50% identity to a nucleotide sequence selected from the group consisting of SEQ ID NO;
14, SEQ ID NO;
15, SEQ ID NO;
16 and SEQ ID NO;
17, SEQ ID NO;
36, SEQ ID NO;
45, and SEQ ID NO;
55;
b) constructing a vector comprising said isolated nucleotide sequence;
c) introducing said vector into a host cell for a time and under conditions sufficient for expression of Δ
4-desaturase; and
d) exposing said expressed Δ
4-desaturase to a substrate polyunsaturated fatty acid in order to convert said substrate to a product polyunsaturated fatty acid. - View Dependent Claims (16, 17, 18, 22, 23, 24, 25, 26, 27, 28)
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19. A method of producing a polyunsaturated fatty acid comprising the steps of:
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a) exposing a substrate polyunsaturated fatty acid to one or more enzymes selected from the group consisting of a desaturase and an elongase in order to convert said substrate to a product polyunsaturated fatty acid; and
b) exposing said product polyunsaturated fatty acid of step (a) to a Δ
4-desaturase comprising an amino acid sequence selected from the group consisting of SEQ ID NO;
18, SEQ ID NO;
19, SEQ ID NO;
20, SEQ ID NO;
21, SEQ ID NO;
37, SEQ ID NO;
46 and SEQ ID NO;
56, in order to convert said product polyunsaturated fatty acid to a final product polyunsaturated fatty acid. - View Dependent Claims (20, 21)
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29. An isolated nucleotide sequence or fragment thereof comprising or complementary to a nucleotide sequence encoding a polypeptide having desaturase activity, wherein the amino acid sequence of said polypeptide has at least 30% identity to the amino acid sequence of SEQ ID NO:
- 56.
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30. An isolated nucleotide sequence or fragment thereof comprising or complementary to a nucleotide sequence having at least 40% identity to the nucleotide sequence of SEQ ID NO:
- 55.
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31. A purified polypeptide which desaturates polyunsaturated fatty acids at carbon 4 and has an amino acid sequence having at least 30% identity to the amino acid sequence of SEQ ID NO:
- 56.
Specification