Δ 15 desaturases suitable for altering levels of polyunsaturated fatty acids in oleaginous plants and yeast
First Claim
Patent Images
1. An isolated nucleic acid fragment encoding a fungal Δ
- 15 desaturase enzyme, selected from the group consisting of;
(a) an isolated nucleic acid fragment encoding the amino acid sequence as set forth in SEQ ID NO;
2;
(b) an isolated nucleic acid fragment that hybridizes over the full length of the isolated nucleic acid fragment of (a) encoding the amino acid sequence of SEQ ID NO;
2 under the following hybridization conditions;
0.1×
SSC, 0.1% SDS, 65°
C. and washed with 2×
SSC, 0.1% SDS followed by 0.1×
SSC, 0.1% SDS; and
an isolated nucleic acid fragment that is complementary over the full length of (a) or (b).
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Abstract
The present invention relates to fungal Δ-15 fatty acid desaturases that are able to catalyze the conversion of linoleic acid (18:2, LA) to alpha-linolenic acid (18:3, ALA). Nucleic acid sequences encoding the desaturases, nucleic acid sequences which hybridize thereto, DNA constructs comprising the desaturase genes, and recombinant host plants and microorganisms expressing increased levels of the desaturases are described. Methods of increasing production of specific omega-3 and omega-6 fatty acids by over-expression of the Δ-15 fatty acid desaturases are also described herein.
61 Citations
34 Claims
-
1. An isolated nucleic acid fragment encoding a fungal Δ
- 15 desaturase enzyme, selected from the group consisting of;
(a) an isolated nucleic acid fragment encoding the amino acid sequence as set forth in SEQ ID NO;
2;(b) an isolated nucleic acid fragment that hybridizes over the full length of the isolated nucleic acid fragment of (a) encoding the amino acid sequence of SEQ ID NO;
2 under the following hybridization conditions;
0.1×
SSC, 0.1% SDS, 65°
C. and washed with 2×
SSC, 0.1% SDS followed by 0.1×
SSC, 0.1% SDS; andan isolated nucleic acid fragment that is complementary over the full length of (a) or (b). - View Dependent Claims (2, 3, 5, 6, 7, 8, 9, 10, 11, 12, 33, 34)
wherein the sequenced genomic fragment encodes a Δ
15 desaturase enzyme.
- 15 desaturase enzyme, selected from the group consisting of;
-
34. The isolated nucleic acid fragment of either of claims 1 or 4 wherein the isolated nucleic acid fragment encodes a polypeptide having both a Δ
- 15 desaturase activity and a Δ
12 desaturase activity.
- 15 desaturase activity and a Δ
-
4. An isolated nucleic acid fragment comprising a first nucleotide sequence encoding a Δ
- 15 desaturase enzyme of at least 402 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide having the sequence as set forth in SEQ ID NO;
2;or a second nucleotide sequence comprising the complement of the first nucleotide sequence.
- 15 desaturase enzyme of at least 402 amino acids that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide having the sequence as set forth in SEQ ID NO;
-
13. A method for the production of α
- -linolenic acid comprising;
a) providing a host cell comprising; (i) an isolated nucleic acid fragment encoding a protein having Δ
15 desaturase activity that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide having the sequence as set forth in SEQ ID NO;
2; and(ii) a source of linoleic acid; b) growing the host cell of step (a) under conditions wherein the nucleic acid fragment encoding a protein having Δ
15 desaturase activity is expressed and the linoleic acid is converted to α
-linolenic acid; andc) optionally recovering the α
-linolenic acid of step (b). - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 28, 29, 30, 31, 32)
- -linolenic acid comprising;
-
14. A method for the production of α
- -linolenic acid comprising;
a) providing a host cell comprising; (i) an isolated nucleic acid fragment encoding a protein having Δ
15 desaturase activity that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide having the sequence as set forth in SEQ ID NO;
2; and(ii) a source of oleic acid; b) growing the host cell of step (a) under conditions wherein the nucleic acid fragment encoding a protein having Δ
15 desaturase activity is expressed and the oleic acid is converted to α
-linolenic acid; andc) optionally recovering the α
-linolenic acid of step (b).
- -linolenic acid comprising;
-
22. A method for the production of ω
- -3 fatty acids in a host cell comprising;
a) providing a host cell comprising; (i) an isolated nucleic acid fragment encoding a protein having Δ
15 desaturase activity that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide having the sequence as set forth in SEQ ID NO;
2; and(ii) genes encoding a functional ω
-3/ω
-6 fatty acid biosynthetic pathway;b) providing a source of desaturase substrate consisting of oleic acid; c) growing the host cell of step (a) with the desaturase substrate of step (b) under conditions wherein ω
-3 fatty acids are produced; andd) optionally recovering the ω
-3 fatty acids of step (c). - View Dependent Claims (25, 26, 27)
- -3 fatty acids in a host cell comprising;
-
23. A method of increasing the ratio of ω
- -3 fatty acids to ω
-6 fatty acids in a host cell producing ω
-3 fatty acids to ω
-6 fatty acids comprising;a) providing a host cell producing ω
-3 fatty acids and ω
-6 fatty acids;b) introducing into the host cell of (a) an isolated nucleic acid fragment encoding a protein having at least 90% identity based on the Clustal method of alignment when compared to a polypeptide having the sequence as set forth in SEQ ID NO;
2, wherein the polypeptide binds both oleic acid and linoleic acid as an enzyme substrate, wherein the ratio of ω
-3 fatty acids to ω
-6 fatty acids are increased. - View Dependent Claims (24)
- -3 fatty acids to ω
Specification