Li Hongxia, Zhu Shaozhou, Zheng Guojun
State Key Laboratory of Chemical Resources Engineering, Beijing University of Chemical Technology, Beijing 100029, People's Republic of China.
State Key Laboratory of Chemical Resources Engineering, Beijing University of Chemical Technology, Beijing 100029, People's Republic of China.
Bioorg Med Chem Lett. 2018 Apr 1;28(6):1071-1076. doi: 10.1016/j.bmcl.2018.02.019. Epub 2018 Feb 10.
Based on bioinformatics analysis, the promiscuous (+)-γ-lactamase activity of an amidase was identified in Rhodococcus erythropolis PR4 and found to be involved in the nitrile hydratase pathway. The amidase is highly enantioselective and can be used in the kinetic resolution of the Vince lactam. The known structure provides a rare insight into the catalytic mechanism of (+)-γ-lactamase with absolute chiral selectivity. This lactamase was cloned, purified, biochemically characterized, and demonstrated to be an ideal catalyst for the preparation of carbocyclic nucleosides of pharmaceutical interest. The chiral selectivity of this enzyme was investigated by molecular docking and site-specific mutagenesis, which provides a foundation for further engineering of these versatile biocatalysts.
基于生物信息学分析，在红平红球菌PR4中鉴定出一种酰胺酶具有混杂的（+）-γ-内酰胺酶活性，并发现其参与腈水合酶途径。该酰胺酶具有高度对映选择性，可用于Vince内酰胺的动力学拆分。已知结构为具有绝对手性选择性的（+）-γ-内酰胺酶的催化机制提供了难得的见解。该内酰胺酶被克隆、纯化、进行了生化表征，并被证明是制备具有药学意义的碳环核苷的理想催化剂。通过分子对接和位点特异性诱变研究了该酶的手性选择性，为进一步改造这些多功能生物催化剂奠定了基础。
