基于大系统优化的CPC酰化酶的计算设计及实验验证

项目名称： 基于大系统优化的CPC酰化酶的计算设计及实验验证

项目编号： No.21276136

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： 朱玉山

作者单位： 清华大学

项目金额： 80万元

中文摘要： 本申请以催化CPC水解制备7-ACA的反应过程为例研究建立计算酶设计的大系统优化模型，并开发求解该模型对应的大规模混合整数线性规划问题的全局优化算法，最后通过实验测定CPC酰化酶突变株的活性检验计算酶设计模型的正确性及对模型中的参数进行修正。本申请在具体的建模过程中将重点研究考虑蛋白质结构模板柔性对酶活性位点处氨基酸序列优化的影响，并将反应路径上酶与底物的一系列不同结合状态考虑进酶设计的模型中，通过模型分析及实验验证建立酶活性位点处的氨基酸序列、复合物结构及活性的定量关系，以此来分析决定酶活性位点处氨基酸序列的基本物理化学规律，为得到高效催化CPC水解反应的CPC酰化酶突变株打下理论基础。基于高效的CPC酰化酶突变株的一步酶法绿色工艺有望在制备医药中间体7－ACA的工业过程中取代现有的化学法及两步酶法，从而达到节能减排的目的，因此本申请在工业应用上也具有重要的推广价值。

中文关键词： 计算酶设计；全局优化；酶催化；分子模拟；头孢菌素酰化酶

英文摘要： In this project, a systematic optimization based computational enzyme design model will be developed in terms of the preparation reaction of 7-ACA which is catalyzed by CPC acylase, and then an exclusive and efficient global optimization algorithm will be identified in order to solve the large scale mixed-integer linear programming problem which corresponds to the computational enzyme design model. Finally, the activities of the designed acylase mutants will be measured to validate the correctness of the proposed model for enzyme design, and the parameters in the model could be further improved by using the quantitative structure-activity relationship. Moreover, the flexibility of the enzyme template and a series of transition states during the reaction coordinate will be incorporated into the computational enzyme model and their effects on the sequence selection in active site will be studied and help to establish the fine relationship between sequence, structure, and function so as to discover the physio-chemical regulation which determined the unique amino-acid sequence in the enzyme active site. The potentially designed CPC acylase is promising to develop a one-step enzymatic green process for 7-ACA preparation, which is an crucial pharmaceutical intermediate for antibiotics production, and this process repl

英文关键词： computational enzyme design；global optimization；enzyme catalysis；molecular simulation；cephalosporin acylase