氧化葡萄糖酸杆菌短链脱氢酶结构-功能解析及其生物催化应用基础研究

项目名称： 氧化葡萄糖酸杆菌短链脱氢酶结构-功能解析及其生物催化应用基础研究

项目编号： No.21276084

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 化学工业

项目作者： 魏东芝

作者单位： 华东理工大学

项目金额： 80万元

中文摘要： 氧化葡萄糖酸杆菌是氧化发酵的创立菌和氧化催化应用最广泛的微生物之一，申请人已揭示该菌若干新酶及其催化的90余种反应（部分成功应用）。前期研究基础表明，对其胞内所含最多成员的短链脱氢酶（SDR）的深入研究在学术和应用方面都具有重要意义。本项目拟对13条SDR基因进行分析及功能预测，结合对其生物催化特征的认知，进一步挖掘其区域选择性和立体选择性，获得可应用于重要生物催化反应的新酶，发掘新反应，揭示其胞内氧化还原的生物学功能。SDR家族庞大，参与催化的反应类型众多，其构效关系研究立于当前学术前沿。申请人拟采用蛋白结晶技术结合计算机动力学模拟，验证SDR功能位点或motif，揭示酶催化共性机制，深入解析SDR的结构与功能。借助于理性设计拓宽底物选择性、提高立体选择性、改进辅酶偏好性，为工业生物催化应用提供3-5种理想的新酶，构建1-2株重组菌作为手性醇等重要化合物拆分或合成的生物催化剂。

中文关键词： 氧化葡萄糖酸杆菌；羰基还原酶；催化性质；立体选择性；分子改造

英文摘要： Gluconobacter oxydans, which developed the oxidative fenmentation, has been widely used in oxidative catalyzation. We have determined a series of new enzymes in G.oxydans as well as about ninety reactions, and several were successfully used in industry field. Based on these researches, both theory and application value of the short-chain dehydrogenases/reductases (SDR) are of great importance. Herein, this proposal would interpret its biological functions by designing to analyze and forecast the 13 SDR genes and deeply explore their regio- and stereo-selectivity which also help to discover the new types of catalyzation and new applications.The SDR superfamily involve in many types of catalytic reaction, the relationship research on protein structure and enzymatic function is the most forward field. Protein crystallization, structural analysis, rational design and direct mutagenesis would provide clues to explore their inner relations and give comprehensive understanding on SDR in G. oxydans. Artificial enzymes which have extended substrate acceptance and coenzyme preference can be further obtained. Finally , 3-5 ideal enzymes and 1 or 2 strains can be provided for synthesis or seperation of chiral alcohol in industrial application area.

英文关键词： Gluconobacter oxydans；carbonyl reductase；Catalytic properties；Stereoselectivity；Molecular engineering