通过定向进化提高环氧水解酶的对映选择性。

Enhancing the enantioselectivity of an epoxide hydrolase by directed evolution.

作者信息

Reetz Manfred T, Torre Claudia, Eipper Andreas, Lohmer Renate, Hermes Marcus, Brunner Birgit, Maichele Andrea, Bocola Marco, Arand Michael, Cronin Annette, Genzel Yvonne, Archelas Alain, Furstoss Roland

机构信息

Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470 Mülheim/Ruhr, Germany, Institut für Pharmakologie und Toxikologie, Universität Würzburg, Versbacher Strasse 9, 97078 Würzburg, Germany.

出版信息

Org Lett. 2004 Jan 22;6(2):177-80. doi: 10.1021/ol035898m.

DOI:10.1021/ol035898m

PMID:14723522

Abstract

[reaction: see text] The epoxide hydrolase (EH) from Aspergillus niger, which shows a selectivity factor of only E = 4.6 in the hydrolytic kinetic resolution of glycidyl phenyl ether, has been subjected to directed evolution for the purpose of enhancing enantioselectivity. After only one round of error-prone polymerase chain reaction (epPCR), enantioselectivity was more than doubled (E = 10.8). The improved mutant enzyme contains three amino acid exchanges, two of which are spatially far from the catalytically active center.

摘要

[反应：见正文] 黑曲霉的环氧水解酶（EH）在缩水甘油基苯基醚的水解动力学拆分中显示出仅E = 4.6的选择性因子，为提高对映选择性而进行了定向进化。仅经过一轮易错聚合酶链反应（epPCR）后，对映选择性就增加了一倍多（E = 10.8）。改进的突变酶包含三个氨基酸交换，其中两个在空间上远离催化活性中心。