Pereira Suzette L, Huang Yung-Sheng, Bobik Emil G, Kinney Anthony J, Stecca Kevin L, Packer Jeremy C L, Mukerji Pradip
Department of Strategic Research, Ross Products Division-Abbott Laboratories, 3300 Stelzer Road, Columbus, OH 43219, USA.
Biochem J. 2004 Mar 1;378(Pt 2):665-71. doi: 10.1042/BJ20031319.
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.
长链n-3多不饱和脂肪酸(PUFAs),如二十碳五烯酸(EPA;20:5 n-3),对人类具有重要的治疗和营养益处。在植物、蓝细菌和线虫中,ω3-去饱和酶催化由n-6脂肪酸前体形成这些n-3脂肪酸。在此,我们描述了从富含EPA的真菌水霉(Saprolegnia diclina)中分离和鉴定的一个基因(sdd17),它编码一种新型的ω3-去饱和酶。该基因是通过使用与已知ω3-去饱和酶保守区域相对应的寡核苷酸引物对水霉cDNA文库进行PCR扩增而分离得到的。在存在各种脂肪酸底物的情况下,该基因在酿酒酵母中的表达表明,重组蛋白能够专门使20碳的n-6脂肪酸底物去饱和,对花生四烯酸(ARA;20:4 n-6)有明显偏好,并将其转化为EPA。这种活性不同于任何生物体中已知的ω3-去饱和酶。植物和蓝细菌的ω3-去饱和酶专门使18碳的n-6多不饱和脂肪酸去饱和,秀丽隐杆线虫的一种ω3-去饱和酶优先使18碳的多不饱和脂肪酸而非20碳的底物去饱和,并且在酵母中表达时不能将ARA转化为EPA。sdd17编码的去饱和酶在转基因体细胞大豆胚中也具有功能,能将外源提供的ARA转化为EPA,从而证明了其在转基因油料作物中用于生产EPA的潜力。