Long-chain n−3 PUFAs (polyunsaturated fatty acids) such as EPA (eicosapentaenoic acid; 20:5n−3) have important therapeutic and nutritional benefits in humans. In plants, cyanobacteria and nematodes, ω3-desaturases catalyse the formation of these n−3 fatty acids from n−6 fatty acid precursors. Here we describe the isolation and characterization of a gene (sdd17) derived from an EPA-rich fungus, Saprolegnia diclina, that encodes a novel ω3-desaturase. This gene was isolated by PCR amplification of an S. diclina cDNA library using oligonucleotide primers corresponding to conserved regions of known ω3-desaturases. Expression of this gene in Saccharomyces cerevisiae, in the presence of various fatty acid substrates, revealed that the recombinant protein could exclusively desaturate 20-carbon n−6 fatty acid substrates with a distinct preference for ARA (arachidonic acid; 20:4n−6), converting it into EPA. This activity differs from that of the known ω3-desaturases from any organism. Plant and cyanobacterial ω3-desaturases exclusively desaturate 18-carbon n−6 PUFAs, and a Caenorhabditis elegans ω3-desaturase preferentially desaturated 18-carbon PUFAs over 20-carbon substrates, and could not convert ARA into EPA when expressed in yeast. The sdd17-encoded desaturase was also functional in transgenic somatic soya bean embryos, resulting in the production of EPA from exogenously supplied ARA, thus demonstrating its potential for use in the production of EPA in transgenic oilseed crops.
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Research Article|
March 01 2004
A novel omega3-fatty acid desaturase involved in the biosynthesis of eicosapentaenoic acid
Suzette L. PEREIRA;
Suzette L. PEREIRA
1
*Department of Strategic Research, Ross Products Division-Abbott Laboratories, 3300 Stelzer Road, Columbus, OH 43219, U.S.A.
1To whom correspondence should be addressed (e-mail [email protected]).
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Yung-Sheng HUANG;
Yung-Sheng HUANG
*Department of Strategic Research, Ross Products Division-Abbott Laboratories, 3300 Stelzer Road, Columbus, OH 43219, U.S.A.
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Emil G. BOBIK;
Emil G. BOBIK
*Department of Strategic Research, Ross Products Division-Abbott Laboratories, 3300 Stelzer Road, Columbus, OH 43219, U.S.A.
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Anthony J. KINNEY;
Anthony J. KINNEY
†Dupont Crop Genetics, Experimental Station, Wilmington, DE 19880, U.S.A.
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Kevin L. STECCA;
Kevin L. STECCA
†Dupont Crop Genetics, Experimental Station, Wilmington, DE 19880, U.S.A.
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Jeremy C. L. PACKER;
Jeremy C. L. PACKER
‡Department of Genomics and Molecular Biology, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL 60064, U.S.A.
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Pradip MUKERJI
Pradip MUKERJI
*Department of Strategic Research, Ross Products Division-Abbott Laboratories, 3300 Stelzer Road, Columbus, OH 43219, U.S.A.
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Publisher: Portland Press Ltd
Received:
August 28 2003
Revision Received:
November 21 2003
Accepted:
November 26 2003
Accepted Manuscript online:
December 03 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2004
2004
Biochem J (2004) 378 (2): 665–671.
Article history
Received:
August 28 2003
Revision Received:
November 21 2003
Accepted:
November 26 2003
Accepted Manuscript online:
December 03 2003
Citation
Suzette L. PEREIRA, Yung-Sheng HUANG, Emil G. BOBIK, Anthony J. KINNEY, Kevin L. STECCA, Jeremy C. L. PACKER, Pradip MUKERJI; A novel omega3-fatty acid desaturase involved in the biosynthesis of eicosapentaenoic acid. Biochem J 1 March 2004; 378 (2): 665–671. doi: https://doi.org/10.1042/bj20031319
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