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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
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
2015
发表期刊Catal. Sci. Technol.
卷号5期号:5页码:2622-2629
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.
文章类型论文
学科领域生命有机化学
DOI10.1039/c5cy00085h
URL查看原文
收录类别SCI
语种英语
WOS记录号WOS:000353641800010
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被引频次:6[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/331003/39503
专题生命有机化学国家重点实验室
通讯作者Yu HL(郁惠蕾); Xu JH(许建和)
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
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GB/T 7714
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.
APA 栾正娇.,李福龙.,窦帅.,陈琦.,周佳海.,...&许建和.(2015).Substrate channel evolution of an esterase for the synthesis of cilastatin.Catal. Sci. Technol.,5(5),2622-2629.
MLA 栾正娇,et al."Substrate channel evolution of an esterase for the synthesis of cilastatin".Catal. Sci. Technol. 5.5(2015):2622-2629.
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