Purification and characterization of fluorinated ketone reductase from Geotrichum candidum NBRC 5767

被引:10
|
作者
Cao, Chen [1 ]
Fukae, Takuro [1 ]
Yamamoto, Takuro [1 ]
Kanamaru, Shuji [1 ]
Matsuda, Tomoko [1 ]
机构
[1] Tokyo Inst Technol, Grad Sch Biosci & Biotechnol, Dept Bioengn, Midori Ku, Yokohama, Kanagawa 227, Japan
关键词
Enzymes; Biocatalysis; Purification; Enantioseparation; Trifluoroacetophenone; Asymmetric reduction; ASYMMETRIC REDUCTION; CARBONYL REDUCTASE; ALCOHOL-DEHYDROGENASE; TRIFLUOROMETHYL; ACETOPHENONES; BIOREDUCTION; ENZYMES;
D O I
10.1016/j.bej.2013.04.005
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
Biocatalysts that can specifically catalyze the transformations of fluorinated compounds are rare. A novel NADPH-dependent fluorinated ketone reductase that reduces trifluoroacetophenone to (S)-trifluorophenylethanol with excellent enantioselectivity (ee >99%) from Geotrichum candidum NBRC 5767 was isolated, purified, and characterized. Through ammonium sulfate fractionation and several chromatographies, this enzyme was purified to 95-fold with 3.5% yield. Gel filtration chromatography with SDS-PAGE revealed this protein to be a dimer of 60 kDa subunits. An investigation of the substrate specificity of the purified enzyme revealed that it was highly selective for ketones containing fluorines at the alpha position. (C) 2013 Elsevier B.V. All rights reserved.
引用
收藏
页码:13 / 16
页数:4
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