Mechanism of flavin transfer and oxygen activation by the two-component flavoenzyme styrene monooxygenase

被引：72

作者：

Kantz, A ^{[1
]}

Chin, F ^{[1
]}

Nallamothu, N ^{[1
]}

Nguyen, T ^{[1
]}

Gassner, GT ^{[1
]}

机构：

[1] San Francisco State Univ, Dept Chem & Biochem, San Francisco, CA 94132 USA

来源：

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS | 2005年 / 442卷 / 01期

关键词：

electron transfer; flavin monooxygenase; epoxidation; enzyme intermediate; redox; pre-steady-state kinetics; transient kinetics; styrene; reductase; singlet oxygen;

D O I：

10.1016/j.abb.2005.07.020

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：

Styrene monooxygenase (SMO) from Pseudomonas putida S 12 is a two-component flavoenzyme composed of the NADH-specific flavin reductase, SMOB, and FAD-specific styrene epoxidase, SMOA. Here, we report the cloning, and expression of native and histidine-tagged versions of SMOA and SMOB and studies of the flavin transfer and styrene oxygenation reactions. In the reductive half-reaction, SMOB catalyzes the two-electron reduction of FAD with a turnover number of 3200 s(-1). Single turnover studies of the reaction of reduced SMOA with substrates indicate the formation of a stable oxygen intermediate with the absorbance characteristics of a flavin hydroperoxide. Based on the results of numerical simulations of the steady-state mechanism of SMO, we find that the observed coupling of NADH and styrene oxidation can be best explained by a model, which includes both the direct transfer and passive diffusion of reduced FAD from SMOB to SMOA. (c) 2005 Elsevier Inc. All rights reserved.

引用

页码：102 / 116

页数：15

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