Directed Evolution as a Means to Optimize and Understand Molecular Biocatalysts

定向进化作为优化和理解分子生物催化剂的一种手段

• 批准号: 5427415
• 负责人: Professor Dr. Manfred T. Reetz
• 金额: --
• 依托单位: Arbeitsgruppe Reetz
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2004
• 资助国家: 德国
• 起止时间: 2003-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5427415?language=en
• 关键词: Directed; Evolution; Means; Optimize; Understand

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.

该项目的目的是应用定向进化方法来创建对映选择性环氧化物水解酶作为有机合成中的催化剂，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（德国维尔茨堡大学）和我们研究所的理论家 Walter Thiel 教授的密切合作是该项目的一个组成部分，他开发了两种所用环氧化物水解酶（黑曲霉和柠檬烯环氧化物水解酶）的表达系统。两者均已在此 DFG-Schwerpunkt 中提交了申请。我们计划通过进行适当的诱变/电子筛选循环，从结构上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，从而从定向进化中学习。