Directed evolution of cytochrome P450 for sterol epoxidation

被引：0
作者：
Dan Jiang
Ran Tu
Peng Bai
Qinhong Wang
机构：
[1] Tianjin University,School of Chemical Engineering and Technology
[2] Tianjin Institute of Industrial Biotechnology,Key Laboratory of Systems Microbial Biotechnology
[3] Chinese Academy of Sciences,undefined
来源：
Biotechnology Letters | 2013年 / 35卷
关键词：
Cytochrome P450; Directed evolution; Epoxidation; Sterol;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
16,17-Epoxysterol plays an important role in pharmaceutical steroid synthesis. To investigate the potential application of cytochrome P450 for epoxysterol synthesis, an approach to the epoxidation of 16,17-epoxysterol, based on directed evolution of cytochrome P450 BM-3, was developed. This comprised random gene mutagenesis for optimizing the activity of P450 BM-3 for epoxidation of hydrophobic sterol, followed by the 7-ethoxycoumarin de-ethylation assay for general enzyme activity detection and the modified picric acid assay for epoxidation activity screening. By the two-step screening, one mutant from 792 clones showed specific substrate activity of converting progesterone to 16,17-epoxysterol, which validated the possibility to evolve the cytochrome P450 for the synthesis of steroidal epoxides.
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页码：1663 / 1668
页数：5
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