Directed Evolution as a Means to Optimize and Understand Molecular Biocatalysts

本项目的目的是应用定向进化的方法，在有机合成中创造作为催化剂的对映选择性环氧化物水解酶，并在分子基础上揭示立体选择性的来源。与生物学家/毒理学家Michael Arand教授博士（德国维尔茨堡大学）和我们自己研究所的理论家Walter Thiel教授博士的密切合作是该项目的一个组成部分，Michael Arand教授开发了两种可用的环氧化物水解酶（尼日尔和柠檬烯环氧化物水解酶）的表达系统。两人都在DFG-Schwerpunkt提交了申请。我们计划从定向进化中学习，通过进行适当的诱变/筛选循环，结构表征最具对映选择性的突变体，并使用分子动力学和量子力学计算进行理论研究，以揭示对映选择性的来源。

The purpose of this project is to apply the methods of directed evolution in the quest to create enantioselective epoxide hydrolases as catalysts in organic synthesis and to uncover the source of stereoselectivity on a molecular basis. Close collaboration with the biologist/toxicologist Prof. Dr. Michael Arand (Universität Würzburg, Germany) who has developed expression systems of the two epoxide hydrolases to be used (Aspergillus niger and limonene epoxide hydrolase) and the theoretician Prof. Dr. Walter Thiel in our own institute is an integral part of the project. Both have submitted applications in this DFG-Schwerpunkt. We plan to learn from directed evolution by performing appropriate cycles of mutagenesis/eescreening, structurally characterizing the most enantioselective mutants and carrying out theoretical studies using molecular dynamics and quantum mechanical calculations in order to uncover the source of enantioselectivity.