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学科主题: 生命有机化学
题名: Reshaping an Enzyme Binding Pocket for Enhanced and Inverted Stereoselectivity: Use of Smallest Amino Acid Alphabets in Directed Evolution
其他题名: 通过最小氨基酸列表使用来定向进化改造酶活性口袋残基以实现立体选择性的提高和反转
作者: Sun ZT(孙周通)1; 理查德·郎斯达勒1; Kong XD(孔旭东)1; Xu JH(许建和)1; Zhou JH(周佳海)1; 瑞兹·曼弗雷1
通讯作者: 周佳海 ; 瑞兹·曼弗雷
刊名: Angew. Chem.-Int. Edit.
发表日期: 2015
DOI: 10.1002/anie.201501809
卷: 54, 期:42, 页:12410-12415
收录类别: SCI
文章类型: 论文
英文摘要: Directed evolution based on saturation mutagenesis at sites lining the binding pocket is a commonly practiced strategy for enhancing or inverting the stereoselectivity of enzymes for use in organic chemistry or biotechnology. However, as the number of residues in a randomization site increases to five or more, the screening effort for 95% library coverage increases astronomically until it is no longer feasible. We propose the use of a single amino acid for saturation mutagenesis at superlarge randomization sites comprising 10 or more residues. When used to reshape the binding pocket of limonene epoxide hydrolase, this strategy, which drastically reduces the search space and thus the screening effort, resulted in R,R- and S,S-selective mutants for the hydrolytic desymmetrization of cyclohexene oxide and other epoxides. X-ray crystal structures and docking studies of the mutants unveiled the source of stereoselectivity and shed light on the mechanistic intricacies of this enzyme.
语种: 英语
相关网址: 查看原文
WOS记录号: WOS:000363397300028
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.sioc.ac.cn/handle/331003/39534
Appears in Collections:生命有机化学国家重点实验室_期刊论文

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作者单位: 1.德国马普研究所
2.德国马尔堡大学
3.华东理工大学
4.中科院上海有机化学研究所, 生命有机化学国家重点实验室

Recommended Citation:
Sun ZT,理查德·郎斯达勒,Kong XD,et al. Reshaping an Enzyme Binding Pocket for Enhanced and Inverted Stereoselectivity: Use of Smallest Amino Acid Alphabets in Directed Evolution[J]. Angew. Chem.-Int. Edit.,2015,54(42):12410-12415.
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