调节中链脱酶/还原酶的立体选择性,通过创建一个额外的活性口袋来容纳亲切的异环基
1Laboratory of Brewing Microbiology and Applied Enzymology, School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, China. ynie@jiangnan.edu.cn.
研究人员设计了一种中链脱酶/还原酶 (MDR) 酶,以控制异环基的立体选择性还原. 这一突破增强了性酒精的合成,这对药物开发至关重要.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 有机合成 有机合成
- 药用化学 医学化学
背景情况:
- 中链脱酶/还原酶 (MDRs) 对于合成药品中使用的性异环醇醇至关重要.
- 通过碳缩酶 (CRs) 对异环基的立体选择性降解缺乏明确的调节机制.
- 修改酶的立体选择性是有效的不对称合成的关键.
研究的目的:
- 阐明和控制使用工程MDRs的异环基的立体选择性降解.
- 开发一种以结构为导向的策略,用于修改MDR的立体选择性.
- 为了提高MDR的催化效率和立体选择性,用于制药构建块.
主要方法:
- 来自 *Spathaspora passalidarum* 的碳基还原酶 (SpCR) 的结构引导酶工程.
- 引入额外的基底结合活性口袋以逆转立体选择性.
- 测试工程突变 (m48) 对12个异环基的催化活性和立体选择性.
主要成果:
- 与野生类型的SpCR.相比,工程突变的m48表现出反向立体选择性.
- 对于12个测试的异环基,m48表现出高的催化活性 (转化率>99%) 和优异的反选择性 (ee值>99%).
- 通过积极的口袋工程来实现MDR立体选择性的成功调节.
结论:
- 设计的活性口袋可以有效调节中链脱酶/还原酶的立体选择性.
- 这种以结构为导向的策略为修改MDR立体选择性提供了一种超越传统方法的新方法.
- 增强的m48突变提供了一个强大的生物催化剂,用于 valuable chiral heterocyclic alcohols 的不对称合成.
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