文献类型：期刊文献

英文题名：Regioselectivity Inversion of an O-Methyltransferase via Semi-rational Mutagenesis Combined with Metal Ion Substitution

作者：Su, Ying[1];Li, Hai-Peng[1];Zhang, Man[1];Ding, Xu-Wei[1];Xu, Jian-He[1];Chen, Qi[1];Zheng, Gao-Wei[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2022

卷号：14

期号：23

外文期刊名：CHEMCATCHEM

收录：;EI(收录号：20224513062667);WOS:【SCI-EXPANDED(收录号:WOS:000874117500001)】；

基金：This work was financially supported by the National Key Research and Development Program of China (2019YFA09005000 and 2021YFC2102800), the National Natural Science Foundation of China (21878085, 31971380, 21472045 and 21871085), and the Fundamental Research Funds for the Central Universities (22221818014).

语种：英文

外文关键词：biocatalysis; metal ion substitution; O-methyltransferase; protein engineering; regioselectivity

摘要：S-adenosyl-L-methionine (SAM)-dependent O-methyltransferases (O-MTs) are a class of valuable and attractive biocatalysts for alkylation of catechols. In this work, an O-methyltransferase (GeOMT) from the plant Galanthus elwesii was identified and characterised. GeOMT methylated a series of catechols with excellent regioselectivity. Moreover, a strategy was developed that combines structure-directed protein engineering and metal ion substitution for regioselective inversion of GeOMT. The resultant W50M/A53N/Y186K triple variant preferentially coordinated Ni2+ instead of Mg2+ and displayed almost complete inversion in regioselectivity from 1 : 99 to 94 : 6 (para/meta) for 4-nitrocatechol, the para-methylated product of which is a vital plant growth regulator. This represents a promising strategy for regioselectivity improvement of O-MTs.