探究黄素在水杨酸羟化酶催化的氧化脱羧反应中的作用机制。
Probing the mechanism of flavin action in the oxidative decarboxylation catalyzed by salicylate hydroxylase.
机构信息
Departamento de Química, Instituto de Ciências Exatas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
Departamento de Química, Instituto de Ciências Exatas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
出版信息
Methods Enzymol. 2023;685:241-277. doi: 10.1016/bs.mie.2023.03.017. Epub 2023 May 16.
Salicylate hydroxylase (NahG) is a FAD-dependent monooxygenase in which the reduced flavin activates O coupled to the oxidative decarboxylation of salicylate to catechol or uncoupled from substrate oxidation to afford HO. This chapter presents different methodologies in equilibrium studies, steady-state kinetics, and identification of reaction products, which were important to understand the SAr mechanism of catalysis in NahG, the role of the different FAD parts for ligand binding, the extent of uncoupled reaction, and the catalysis of salicylate's oxidative decarboxylation. These features are likely familiar to many other FAD-dependent monooxygenases and offer a potential asset for developing new tools and strategies in catalysis.
水杨酸羟化酶（NahG）是一种依赖黄素腺嘌呤二核苷酸（FAD）的单加氧酶，其中还原型黄素通过与水杨酸的氧化脱羧反应偶联，将 O 激活为儿茶酚，或者与底物氧化脱耦，生成 HO。本章介绍了平衡研究、稳态动力学和反应产物鉴定的不同方法，这些方法对于理解 NahG 中水杨酸羟化酶的 SAr 催化机制、不同 FAD 部分在配体结合中的作用、脱耦反应的程度以及水杨酸的氧化脱羧催化作用非常重要。这些特征可能为许多其他依赖 FAD 的单加氧酶所熟悉，并为开发催化作用的新工具和策略提供了潜在的优势。
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