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从梭鱼(Muraenesox cinereus)中分离并功能表征 delta 6-去饱和酶基因。

Isolation and functional characterization of a delta 6-desaturase gene from the pike eel (Muraenesox cinereus).

机构信息

National Academy of Agricultural Science, Rural Development Administration, Suwon, 441-707, Republic of Korea.

出版信息

J Microbiol. 2013 Dec;51(6):807-13. doi: 10.1007/s12275-013-3144-3. Epub 2013 Oct 5.

Abstract

Stearidonic acid (STA; 18:4n-3) and γ-linolenic acid (GLA; 18:3n-6) are significant intermediates in the biosynthetic pathway for the very-long-chain polyunsaturated fatty acids of eicosapentaenoic acid (EPA; 20:5n-3) and arachidonic acid (ARA; 20:4n-6), respectively. To develop a sustainable system for the production of dietary polyunsaturated fatty acids, we focused on the action of the enzyme delta 6-desaturase (D6DES) on the essential acids, linoleic acid (LA; 18:2n-6) and α-linolenic acid (ALA; 18:3n-3). A 1,335-bp full-length cDNA encoding D6DES (McD6DES) was cloned from Muraenesox cinereus using degenerate PCR and RACE-PCR methods. To investigate the enzymatic activity of McD6DES in the production of n-6 and n-3 fatty acids, a recombinant plasmid expressing McD6DES (pYES-McD6DES) was transformed into and expressed in Saccharomyces cerevisiae. The exogenously expressed McD6DES produced GLA and STA at conversion rates of 14.2% and 45.9%, respectively, from the exogenous LA and ALA substrates. These results indicate that McD6DES is essentially a delta 6-desaturase involved in very-long-chain polyunsaturated fatty acid synthesis.

摘要

硬脂烯酸(STA;18:4n-3)和γ-亚麻酸(GLA;18:3n-6)分别是二十碳五烯酸(EPA;20:5n-3)和花生四烯酸(ARA;20:4n-6)的非常长链多不饱和脂肪酸生物合成途径中的重要中间体。为了开发一种可持续的生产膳食多不饱和脂肪酸的系统,我们专注于 delta 6-去饱和酶(D6DES)对必需脂肪酸亚油酸(LA;18:2n-6)和α-亚麻酸(ALA;18:3n-3)的作用。我们使用简并 PCR 和 RACE-PCR 方法从灰海鳗(Muraenesox cinereus)中克隆了编码 D6DES(McD6DES)的全长 1335bp cDNA。为了研究 McD6DES 在生产 n-6 和 n-3 脂肪酸中的酶活性,我们将表达 McD6DES 的重组质粒(pYES-McD6DES)转化并表达在酿酒酵母中。外源表达的 McD6DES 分别从外源性 LA 和 ALA 底物中将 GLA 和 STA 的转化率提高到 14.2%和 45.9%。这些结果表明 McD6DES 本质上是一种参与非常长链多不饱和脂肪酸合成的 delta 6-去饱和酶。

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