P450酶特性研究与新型杂合酶的构建  

Survey of P450s and construction of novel hybrid enzyme

作　　者：张玲玲[1] 巴丽娜[1] 林章凛[1] 朱玉山[1] 

机构地区：[1]清华大学化学工程系,北京100084

出　　处：《计算机与应用化学》2010年第1期103-107,共5页Computers and Applied Chemistry

基　　金：国家高技术研究发展计划(863)资助项目(2008AA02Z208)

摘　　要：细胞色素P450酶是一类在自然界中广泛存在的含有亚铁血红素的酶系,能够催化以小分子有机物单加氧反应为主的多种生物化学反应,在药物生产和人类健康等领域发挥着重要的作用。细胞色素P450酶催化过程的核心是辅基血红素HEME中铁离子与氧气分子的反应,涉及2个电子转移的步骤。经由氧化还原辅酶的电子传递成为P450酶催化循环的前提,该酶也因和氧化还原辅酶耦合方式的不同而分为3类。作为1个电子自给自足的多域细胞色素P450,P450BM3能够将NAD(P)H产生的电子100%用于产物生成,其高效的催化性能为研究P450酶结构功能关系和电子传递机制提供了优良模板。仿照P450BM3构建P450杂合酶是近年来细胞色素P450酶研究领域的热点之一。本文总结了细胞色素P450酶的催化过程和分类,以及P450BM3酶的界面、连接肽、电子传递和活性位点的相关研究成果,并简要介绍了细胞色素P450杂合酶的人工构建。我们依据蛋白质系统优化的方法,将理性设计与实验验证相结合,从事构建和改善细胞色素P450sca-2杂合酶,以期通过酶法转变调血脂药物普伐他汀生产过程中转化效率低且成本高的现状,具有重要的应用价值。Cytochrome P450 proteins belong to a universal heme contained enzyme superfamily which can catalyze an extremely diverse number of reactions,typically the monooxygenase reactions of small organic molecules.Cytochrome P450s bear great importance in the areas of pharmaceutical products and human health.The key process of P450 catalytic cycle is the reaction between the Fe ion of the heme and the oxygen molecule,which involves two electron transfer steps.Electron transfer from redox partners to the P450s is a precondition of P450 catalytic cycle.Depending on the nature of the redox partner,P450 enzymes generally fall into three broad classes.As a multi-domain self-sufficient protein,P450BM3 is capable of making use of 100%electrons from NAD（P）H in the product generation process.With the unprecedented Catalytic ability,P450BM3 provides researchers with an excellent model for studying structure-function relationships in P450s and the mechanism of electron transfer.The construction of hybrid enzymes imitating P450BM3 has become one of the hot topics in recent years.In this article,we summarized the catalytic process and classification of P450 enzymes,reviewed the relevant studies of the interface,the linker,the electron transfer and the active sites of P450BM3,and briefly introduced the design of cytochrome P450fusion enzymes.We focus on constructing and engineering of hybrid P450sca-2,with both rational design and experimental verification employing the methods of protein system optimization.This enzyme will contribute much to the manufacturing of Pravastatin,an important cholesterol-lowering drug.

关 键 词：细胞色素P450 P450BM3 P450杂合酶 电子传递 综述 

分 类 号：TQ033[化学工程]