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酵母Δ12脂肪酸去饱和酶:基因克隆、表达及功能

Yeast Delta 12 fatty acid desaturase: gene cloning, expression, and function.

作者信息

Watanabe Kyoko, Oura Takahiro, Sakai Hiromichi, Kajiwara Susumu

机构信息

Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Kanagawa, Japan.

出版信息

Biosci Biotechnol Biochem. 2004 Mar;68(3):721-7. doi: 10.1271/bbb.68.721.

Abstract

In an effort to elucidate the molecular mechanisms of fatty acid desaturation in yeast, a complete gene encoding Delta 12 fatty acid desaturase of Saccharomyces kluyveri was cloned. The open reading frame of this gene (named Sk-FAD2) consists of 1,251 bp, encoding 416 amino acids. The deduced Sk-FAD2 protein had 37-55% identity with those from other filamentous fungi. Unlike the genes of these other fungi, S. cerevisiae expressing Sk-FAD2 was found to be capable of synthesizing the dienoic fatty acid hexadecadienoic acid as well as linoleic acid. Moreover, the Sk-FAD2-disrupted strain of S. kluyveri was unable to produce polyunsaturated fatty acids. These results suggested that Sk-FAD2 protein is a unique Delta 12 fatty acid desaturase in S. kluyveri. Analysis of transcriptional expression revealed that Sk-FAD2 was not repressed by exogenous unsaturated fatty acids but responded to low-temperature stress.

摘要

为了阐明酵母中脂肪酸去饱和作用的分子机制,克隆了克鲁维酵母中一个编码Δ12脂肪酸去饱和酶的完整基因。该基因(命名为Sk-FAD2)的开放阅读框由1251个碱基对组成,编码416个氨基酸。推导的Sk-FAD2蛋白与其他丝状真菌的蛋白具有37%-55%的同一性。与这些其他真菌的基因不同,发现表达Sk-FAD2的酿酒酵母能够合成二烯脂肪酸十六碳二烯酸以及亚油酸。此外,克鲁维酵母的Sk-FAD2基因破坏菌株无法产生多不饱和脂肪酸。这些结果表明Sk-FAD2蛋白是克鲁维酵母中一种独特的Δ12脂肪酸去饱和酶。转录表达分析表明,Sk-FAD2不受外源不饱和脂肪酸的抑制,但对低温胁迫有反应。

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