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.
- arachidonic acid
- eicosapentaenoic acid
- fatty acid
- transgenic oil
The nucleotide sequence reported in this paper has been submitted to the DDBJ, EMBL, GenBank® and GSDB Nucleotide Sequence Databases under the accession number AY373823.
Abbreviations used: PUFA, polyunsaturated fatty acid; ARA, arachidonic acid; EPA, eicosapentaenoic acid; LA, linoleic acid; ALA, α-linolenic acid; DGLA, dihomo-γ-linolenic acid; ER, endoplasmic reticulum.
- The Biochemical Society, London ©2004