Saito T, Ochiai H
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo, Japan.
Eur J Biochem. 1999 Oct;265(2):809-14. doi: 10.1046/j.1432-1327.1999.00789.x.
cDNA fragments putatively encoding amino acid sequences characteristic of the fatty acid desaturase were obtained using expressed sequence tag (EST) information of the Dictyostelium cDNA project. Using this sequence, we have determined the cDNA sequence and genomic sequence of a desaturase. The cloned cDNA is 1489 nucleotides long and the deduced amino acid sequence comprised 464 amino acid residues containing an N-terminal cytochrome b5 domain. The whole sequence was 38.6% identical to the initially identified Delta5-desaturase of Mortierella alpina. We have confirmed its function as Delta5-desaturase by over expression mutation in D. discoideum and also the gain of function mutation in the yeast Saccharomyces cerevisiae. Analysis of the lipids from transformed D. discoideum and yeast demonstrated the accumulation of Delta5-desaturated products. This is the first report concering fatty acid desaturase in cellular slime molds.
利用盘基网柄菌cDNA项目的表达序列标签(EST)信息,获得了推测编码脂肪酸去饱和酶特征性氨基酸序列的cDNA片段。利用该序列,我们确定了一种去饱和酶的cDNA序列和基因组序列。克隆的cDNA长1489个核苷酸,推导的氨基酸序列由464个氨基酸残基组成,包含一个N端细胞色素b5结构域。整个序列与最初鉴定的高山被孢霉Δ5-去饱和酶有38.6%的同一性。我们通过在盘基网柄菌中的过表达突变以及在酿酒酵母中的功能获得性突变,证实了其作为Δ5-去饱和酶的功能。对转化的盘基网柄菌和酵母中的脂质分析表明,Δ5-去饱和产物有所积累。这是关于细胞黏菌中脂肪酸去饱和酶的首次报道。