Chromogenic nitrophenolate-based substrates for light-driven hybrid P450 BM3 enzyme assay

被引：17

作者：

Quan Lam ^{[1
]}

Cortez, Alejandro ^{[1
]}

Thanh Truc Nguyen ^{[1
]}

Kato, Mallory ^{[1
]}

Cheruzel, Lionel ^{[1
]}

机构：

[1] San Jose State Univ, Dept Chem, One Washington Sq, San Jose, CA 95192 USA

来源：

JOURNAL OF INORGANIC BIOCHEMISTRY | 2016年 / 158卷

基金：

美国国家科学基金会;

关键词：

Light-driven biocatalysis; Hybrid P450 BM3 enzymes; Colorimetric assay; p-Nitrophenoxy derivatives; Chromogenic substrates; MEDIATED ELECTRON-TRANSFER; LABORATORY EVOLUTION; DIRECTED EVOLUTION; SELF-SUFFICIENT; CYTOCHROME-P450; BIOCATALYST; ACID; HYDROXYLATION; OXIDATION; DISCOVERY;

D O I：

10.1016/j.jinorgbio.2015.12.005

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The incorporation of a p-nitrophenoxy moiety in substrates has enabled the development of colorimetric assays to rapidly screen for O-demethylation activity of P450 enzymes. For the light-driven hybrid P450 BM3 enzymes, where a Ru(II) photosensitizer powers the enzyme upon visible light irradiation, we have investigated a family of p-nitrophenoxy derivatives as useful chromogenic substrates compatible with the light-driven approach. The validation of this assay and its adaptability to a 96-well plate format will enable the screening of the next generation of hybrid P450 BM3 enzymes towards C-I-I bond functionalization of non-natural substrates. (c) 2015 Elsevier Inc. All rights reserved.

引用

页码：86 / 91

页数：6

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