Watanabe Kyoko, Oura Takahiro, Sakai Hiromichi, Kajiwara Susumu
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Kanagawa, Japan.
Biosci Biotechnol Biochem. 2004 Mar;68(3):721-7. doi: 10.1271/bbb.68.721.
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.
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。