离子液体体系中环氧化物水解酶的糖基修饰对其催化性能的调控机制

A epoxide hydrolase screened from Aspergillus niger ZJB-09173 by our team was selected as research object in this project. Based on the significant influence of glycosylation modification on the substrate specificity and stereoselectivity of this enzyme, the epoxide hydrolases with different glycosylation sructure (glycosylation site, glycans types and number) were prepared. The pure solvent, cosolvent, biphaseHeterogebeous systems containing ionic liquid were also prepared. epoxide hydrolases with different glycosylation sructure (glycosylation site, glycans types and number) were prepared.The enzymatic properties and kinetic characteristics of epoxide hydrolases with different glycosylation modification were investigatied in ionic liquid system. The conformation of glycosylated enzyme and its thermodynamic parameter in ionic liquid system were also analyzed. The relations between glycosylation and catalytic function of epoxide hydrolase and the related mechanism were elucidated. The aim of this research is to lay the new ideal and method for the directed restruction of enzymes based on glycosylation modification and ionic liquid system.

以课题组前期自行筛选的Aspergillus niger ZJB-09173环氧化物水解酶为对象，针对在离子液体中糖基修饰与无糖基修饰环氧化物水解酶在底物特异性、立体选择性存在显著差异的现象，通过构建环氧化物水解酶糖基修饰多样性（修饰位点、糖基种类、大小）文库，建立含离子液体纯溶剂、共溶剂、双相和多相体系，研究离子液体体系中糖基修饰环氧化物水解酶的催化性质及反应动力学，分析和计算糖基修饰环氧化物水解酶的构象变化及其折叠热力学参数，探讨离子液体体系中糖基修饰与酶蛋白的相互作用关系，阐明离子液体体系中环氧化物水解酶的糖基修饰对其催化性能调控的分子机制与基本规律，进而为离子液体高效催化体系构建、酶分子的定向改造提供新思路和新方法。

本项目以课题组前期自行筛选的环氧化物水解酶为对象，构建了环氧化物水解酶糖基修饰多样性库，获得了不同修饰位点、糖基种类、大小的糖基修饰环氧化物水解酶。构建了离子液体纯溶剂、共溶、双相等反应体系，研究了离子液体体系中糖基修饰环氧化物水解酶的催化性质及反应动力学，分析和计算糖基修饰环氧化物水解酶的结构与热力学参数变化，探讨离子液体体系中糖基修饰与酶蛋白的相互作用关系，初步阐明离子液体体系中环氧化物水解酶的糖基修饰对其催化性能调控的基本规律。本项目的研究为环氧化物水解酶离子液体高效催化体系构建、环氧化物水解酶分子的功能改造提供新思路和新方法。