藻毒素降解酶MlrA活性中心及其催化降解藻毒素分子机理的研究

项目名称： 藻毒素降解酶MlrA活性中心及其催化降解藻毒素分子机理的研究

项目编号： No.21472061

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 环境化学

项目作者： 冯玲玲

作者单位： 华中师范大学

项目金额： 85万元

中文摘要： 藻毒素降解酶（Microcystinase）是近年发现的能够彻底降解藻毒素，降低藻毒素毒性的多酶复合物。其中藻毒素降解酶A(MlrA)是该多酶复合物在藻毒素解毒、去毒过程中的第一个酶，也是最为关键的一个酶，它可将环状藻毒素降解为线状藻毒素，使藻毒素毒性降低近160倍。本项目拟综合运用基因克隆表达、生物信息学分析、肽段切除及修饰、分子定向进化、计算机辅助分子设计及光谱分析等技术，以藻毒素为小分子探针，以MlrA为靶标酶，系统深入地研究MlrA的结构及其与藻毒素之间相互作用的机理，探讨MlrA活性中心及其催化降解藻毒素的分子机理，以获得准确可靠的酶-配体相互作用模型，确定酶催化的基本反应途径。在此基础上，对MlrA进行设计及定向改造的研究，可望获得高效、稳定、广谱的具有应用前景的藻毒素降解酶，为我国开发具有自主知识产权的新型藻毒素降解酶的创制工作提供合理的设计指导和前期研发基础。

中文关键词： 分子机理；藻毒素降解酶；小分子探针；合理设计；构效关系

英文摘要： Microcystinase multienzyme complex can completely degrade microcystins（MCs）, which has been found in the recent years. Amongst the microcystinase multienzyme complex, MlrA is the first enzyme, which degrades the cycle-microcystins into line-microcystins and toxic of the line- microcystins is decreased 160-fold than that of cycle-microcystins. In this project, we plan to go on a systemic and deep research on the interaction between MlrA and MCs using the MCs as small molecule probes by gene clone and expression,bioinformation engineering,computer model technique combined with spectrum analysis and molecule orthogenesis and so on, and explore its active sites and molecule detoxicated mechanism, in order to gain the accurate and reliable interaction model of enzyme-ligand and confirm fundamental catalyzed pathway and to find out rate-determining step. Based on this research, we will do some research on the design and screen and rebuilt of MlrA and hope to gain new MlrA with high activity and broad spectrum and promising prospect.This work will provide reasonable design guidance and necessary research base for the creation of new MlrA which possess proprietary intellectual property rights.

英文关键词： catalyzed molecular mechanism;microcystinase;small molecular probe;rational design;structure-activity relationship