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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
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
2015
发表期刊Angew. Chem.-Int. Edit.
卷号54期号:42页码:12410-12415
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.
文章类型论文
学科领域生命有机化学
DOI10.1002/anie.201501809
URL查看原文
收录类别SCI
语种英语
WOS记录号WOS:000363397300028
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被引频次:25[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.sioc.ac.cn/handle/331003/39534
专题生命有机化学国家重点实验室
通讯作者Zhou JH(周佳海); 瑞兹·曼弗雷
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
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GB/T 7714
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.
APA 孙周通,理查德·郎斯达勒,孔旭东,许建和,周佳海,&瑞兹·曼弗雷.(2015).Reshaping an Enzyme Binding Pocket for Enhanced and Inverted Stereoselectivity: Use of Smallest Amino Acid Alphabets in Directed Evolution.Angew. Chem.-Int. Edit.,54(42),12410-12415.
MLA 孙周通,et al."Reshaping an Enzyme Binding Pocket for Enhanced and Inverted Stereoselectivity: Use of Smallest Amino Acid Alphabets in Directed Evolution".Angew. Chem.-Int. Edit. 54.42(2015):12410-12415.
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