Functional expression of mammalian NADPH-cytochrome P450 oxidoreductase on the cell surface of Escherichia coli

被引:26
作者
Yim, Sung-Kun
Jung, Heung-Chae
Pan, Jae-Gu [1 ]
Kang, Hyung-Sik
Ahn, Taeho
Yun, Chul-Ho
机构
[1] Chonnam Natl Univ, Hormone Res Ctr, Kwangju 500757, South Korea
[2] Chonnam Natl Univ, Sch Biol Sci & Technol, Kwangju 500757, South Korea
[3] Korea Res Inst Biosci & Biotechnol, GenoFocus Inc, Natl Res Lab Microbial Display, Taejon 305333, South Korea
[4] Korea Res Inst Biosci & Biotechnol, Syst Microbiol Res Ctr, Taejon 305333, South Korea
[5] Chonnam Natl Univ, Coll Vet Med, Dept Biochem, Kwangju 500757, South Korea
关键词
diflavin-containing mammalian enzyme; oxidoreductase; reduction; surface display; whole-cell biocatalyst; ICE-NUCLEATION PROTEIN; ANTLEY-BIXLER-SYNDROME; DISORDERED STEROIDOGENESIS; PSEUDOMONAS-SYRINGAE; ELECTRON-TRANSFER; P-450; REDUCTASE; DISPLAY; ENZYME; CYTOTOXICITY; PURIFICATION;
D O I
10.1016/j.pep.2006.05.013
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
To develop a whole-cell oxidoreductase system without the practical limitation of substrate/product transport, easy preparation, stability of enzymes, and low expression levels, we here report the development of a whole cell biocatalyst displaying rat NADPH-cytochrome P450 oxidoreductase (CPR, 77-kDa) on the surface of Escherichia coli by using ice-nucleation protein from Pseudomonas syringae. Surface localization and functionality of the CPR were verified by flow cytometry, electron microscopy, and measurements of enzyme activities. The results of this study comprise the first report of microbial cell-surface display of diflavin-containing mammalian enzymes. This system will allow us to select and develop oxidoreductases, containing bulky and complex prosthetic groups of FAD and FMN, into practically useful whole-cell blocatalysts for broad biological and biotechnological applications including the selective synthesis of new chemicals and pharmaceuticals, bioconversion, bioremediation, and bio-chip development. (c) 2006 Elsevier Inc. All rights reserved.
引用
收藏
页码:292 / 298
页数:7
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