A novel omega3-fatty acid desaturase involved in the biosynthesis of eicosapentaenoic acid.
Long-chain n-3 PUFAs (polyunsaturated fatty acids) such as EPA (eicosapentaenoic acid; 20:5 n-3) have important therapeutic and nutritional benefits in humans. In plants, cyanobacteria and nematodes, omega3-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 omega3-desaturase. This gene was isolated by PCR amplification of an S. diclina cDNA library using oligonucleotide primers corresponding to conserved regions of known omega3-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; 20:4 n-6), converting it into EPA. This activity differs from that of the known omega3-desaturases from any organism. Plant and cyanobacterial omega3-desaturases exclusively desaturate 18-carbon n-6 PUFAs, and a Caenorhabditis elegans omega3-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.
Pereira
Suzette L
SL
Department of Strategic Research, Ross Products Division-Abbott Laboratories, 3300 Stelzer Road, Columbus, OH 43219, USA. suzette.pereira@abbott.com
Huang
Yung-Sheng
YS
Bobik
Emil G
EG
Kinney
Anthony J
AJ
Stecca
Kevin L
KL
Packer
Jeremy C L
JC
Mukerji
Pradip
P
D000595:Amino Acid Sequence; D001095:Arachidonic Acids / Q000378:metabolism; D015118:Eicosapentaenoic Acid / Q000096:biosynthesis*; D004622:Embryo, Mammalian / Q000378:metabolism; D004625:Embryo, Nonmammalian; D044943:Fatty Acid Desaturases / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D005227:Fatty Acids / Q000032:analysis; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D010802:Phylogeny; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D044762:Saprolegnia / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000196:embryology / Q000378:metabolism
D001095:Arachidonic Acids; D005227:Fatty Acids; D015118:Eicosapentaenoic Acid; D044943:Fatty Acid Desaturases; C078565:omega-3 fatty acid desaturase