Substrate channel evolution of an esterase for the synthesis of cilastatin
文献类型：期刊论文
| 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 |
| 刊名 | Catal. Sci. Technol.
 |
| 出版日期 | 2015 |
| 卷号 | 5期号:5页码:2622-2629 |
| 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 |
| 通讯作者 | 郁惠蕾 ; 许建和 |
| 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. |
| 学科主题 | 生命有机化学 |
| 收录类别 | SCI |
| 原文出处 | http://dx.doi.org/10.1039/c5cy00085h |
| 语种 | 英语 |
| WOS记录号 | WOS:000353641800010 |
| 源URL | [http://ir.sioc.ac.cn/handle/331003/39503]  |
| 专题 | 上海有机化学研究所_生命有机化学国家重点实验室 |
| 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 |
| 推荐引用方式 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. |
入库方式： OAI收割
来源：上海有机化学研究所
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