Δ 5 desaturase and its use in making polyunsaturated fatty acids
AssigneeE.I. du Pont de Nemours and Company
InventorsHoward Glenn Damude, Dana M. Walters Pollak, Zhixiong Xue, Quinn Qun Zhu
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Abstract
The present invention relates to a Δ5 desaturase, which has the ability to convert dihomo-γ-linolenic acid (DGLA; 20:3 ω-6) to arachidonic acid (ARA; 20:4 ω-6) and/or eicosatetraenoic acid (ETA; 20:4 ω-3) to eicosapentaenoic acid (EPA; 20:5 ω-3). Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding Δ5 desaturase along with a method of making long chain polyunsaturated fatty acids (PUFAs) using this Δ5 desaturase in oleaginous yeast are disclosed.
Claims17
An isolated nucleic acid molecule selected from the group consisting of:
- (a) an isolated nucleotide sequence encoding a Δ5 desaturase enzyme as set forth in SEQ ID NO:2;
- (b) an isolated nucleotide sequence that hybridizes with (a) under the following hybridization conditions:
- 0.1×SSC, 0.1% SDS, 65° C. and washed with 2×SSC, 0.1% SDS, wherein said nucleotide sequence encodes a delta-5 desaturase followed by 0.1×SSC, 0.1% SDS;
- and,
- (c) an isolated nucleotide sequence that is completely complementary to (a) or (b).
An isolated nucleic acid molecule comprising a first nucleotide sequence encoding a Δ5 desaturase enzyme of at least 449 amino acids that has at least 95% identity based on BLASTP algorithms 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.
A method for the production of arachidonic acid comprising:
- a) providing a host cell comprising:
- (i) an isolated nucleotide molecule encoding a Δ5 desaturase polypeptide having at least 95% identity when compared to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:2, based on BLASTP algorithms;
- and,
- (ii) a source of dihomo-γ-linolenic acid;
- b) growing the host cell of step (a) under conditions wherein the nucleic acid fragment encoding the Δ5 desaturase polypeptide is expressed and the dihomo-γ-linolenic acid is converted to arachidonic acid;
- and,
- c) optionally recovering the arachidonic acid of step (b).
A method for the production of eicosapentaenoic acid comprising:
- a) providing a host cell comprising:
- (i) an isolated nucleotide molecule encoding a Δ5 desaturase polypeptide having at least 95% identity when compared to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:2, based on BLASTP algorithms;
- and,
- (ii) a source of eicosatetraenoic acid;
- b) growing the host cell of step (a) under conditions wherein the nucleic acid fragment encoding the Δ5 desaturase polypeptide is expressed and the eicosatetraenoic acid is converted to eicosapentaenoic acid;
- and,
- c) optionally recovering the eicosapentaenoic acid of step (b).
Dependencies are inferred from the claim text.