Characterization of a Squalene Synthase from the Thraustochytrid Microalga Aurantiochytrium sp KRS101

被引:24
|
作者
Hong, Won-Kyung [1 ]
Heo, Sun-Yeon [1 ]
Park, Hye-Mi [1 ]
Kim, Chul Ho [1 ]
Sohn, Jung-Hoon [2 ]
Kondo, Akihiko [3 ]
Seo, Jeong-Woo [1 ]
机构
[1] KRIBB, Appl Microbiol Res Ctr, Biomat Res Inst, Jeonbuk 580185, South Korea
[2] KRIBB, Syst & Synthet Biol Res Ctr, Taejon 305333, South Korea
[3] Kobe Univ, Dept Chem Sci & Engn, Grad Sch Engn, Kobe, Hyogo 6578501, Japan
基金
新加坡国家研究基金会;
关键词
Aurantiochytrium; squalene synthase; gene analysis; functional characterization; CLONING; OPTIMIZATION; 18W-13A; GENE;
D O I
10.4014/jmb.1212.12023
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The gene encoding squalene synthase (SQS) of the lipid-producing heterotrophic microalga Aurantiochytrium sp. KRS101 was cloned and characterized. The krsSQS gene is 1,551 bp in length and has two exons and one intron. The open reading frame of the gene is 1,164 bp in length, yielding a polypeptide of 387 predicted amino acid residues with a molecular mass of 42.7 kDa. The deduced krsSQS sequence shares at least four conserved regions known to be required for SQS enzymatic activity in other species. The protein, tagged with His(6), was expressed into soluble form in Escherichia coli. The purified protein catalyzed the conversion of farnesyl diphosphate to squalene in the presence of NADPH and Mg2+. This is the first report on the characterization of an SQS from a Thraustochytrid microalga.
引用
收藏
页码:759 / 765
页数:7
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