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学科主题: 生命有机化学
题名: Substrate channel evolution of an esterase for the synthesis of cilastatin
其他题名: 通过底物通道进化方法使酯酶适合西司他汀的合成
作者: Luan ZJ(栾正娇)1; Li FL(李福龙)1; Dou S(窦帅)1; Chen Q(陈琦)1; Zhou JH(周佳海)1; Yu HL(郁惠蕾)1; Xu JH(许建和)1
通讯作者: 郁惠蕾 ; 许建和
刊名: Catal. Sci. Technol.
发表日期: 2015
DOI: 10.1039/c5cy00085h
卷: 5, 期:5, 页:2622-2629
收录类别: SCI
文章类型: 论文
英文摘要: The esterase RhEst1 from Rhodococcus sp. ECU1013 has been reported for the enantioselective hydrolysis of ethyl (S)-(+)-2,2-dimethylcyclopropane carboxylate, producing the building block of cilastatin. In this work, error-prone PCR and site-directed saturation mutagenesis were applied to RhEst1 for activity improvement, with the pH-indicator assay as a high-throughput screening method. As a result, RhEst1(A147I/V148F/G254A), with mutations surrounding the substrate access channel, showed a 5-fold increase in its specific activity compared with the native enzyme, as well as a 4-fold increase in protein solubility. Combined with the determination of protein structures and computational analysis, this work shows that the amino acids around the substrate channel play a more important role in the activity evolution of RhEst1 than those in the active site.
语种: 英语
相关网址: 查看原文
WOS记录号: WOS:000353641800010
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内容类型: 期刊论文
URI标识: http://ir.sioc.ac.cn/handle/331003/39503
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.华东理工大学
2.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:
Luan ZJ,Li FL,Dou S,et al. Substrate channel evolution of an esterase for the synthesis of cilastatin[J]. Catal. Sci. Technol.,2015,5(5):2622-2629.
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