“Fishes-in-net” 酵母孢子微胶囊式近平滑假丝酵母SCRII酶有机相高效手性合成机制研究

项目名称： “Fishes-in-net” 酵母孢子微胶囊式近平滑假丝酵母SCRII酶有机相高效手性合成机制研究

项目编号： No.31670070

项目类型： 面上项目

立项/批准年度： 2017

项目学科： 生物科学

项目作者： 张荣珍

作者单位： 江南大学

项目金额： 25万元

中文摘要： 近平滑假丝酵母CCTCC M203011是一株高效转化手性苯乙二醇，具有自主知识产权的优良菌株。其立体选择性氧化还原酶SCRII催化不对称反应底物浓度低且专一性强（Bioresour Technol2011,102:483-89；PLoS ONE,2013, 8:e83586；Microb Cell Fact,2012,11:167-75）。本研究拟构建孢子微胶囊酶，该酶采用“Fishes-in-net”催化模式，底物和辅酶自由进出孢子，酶固定于孢子内。研究微胶囊酶催化立体异构反应的生理生化与动力学特性、立体选择性、及酶与辅酶协同和稳定催化的作用模式，为获得稳定的高选择性新立体选择性还原酶有机相高效手性合成提供理论基础。

中文关键词： 孢子微胶囊羰基还原酶；分子手性催化机制；有机相；构效关系；辅因子工程

英文摘要： Candida parapsilosis CCTCC M203011 is an excellent strain for chiral alcohol production with independent intellectual property rights. The stereoselective oxidoreductase SCRII catalyzes asymmetric reaction towards low-concentrated substrate and exhibits strong substrate specificity (Bioresour Technol, 2011, 102:483-89; PLoS ONE, 2013, 8:e83586; Microb Cell Fact, 2012, 11:167-75). The project focuses on construction of spore-encapsulation in Saccharomyces cerevisiae with osw1 gene delection and enhanced permeability. The spore-encapsulation follows the “Fishes-in-net” model: the substrates and cofactors can smoothly get into and out of the spores, while enzyme is immobilized in spores. The physiological, biochemical and dynamic characteristics, substrate and product stereroselectivity, and interaction model between encapsulated enzyme and cofactor will be studied. The cofactor-regeneration metabolic pathways will be strengthened in the yeast-spore encapsulation system. The project will provide theoretical basis of novel and stable carbonyl reductases with high stereoselectivity for efficient chiral synthesis in organic phase.

英文关键词： Spore-encapulated carbonyl reductase;molecular chiral catalysis mechanism;Organic phase;Structure-function relationship;Cofactor engineering