Directed Evolution of RebH for Catalyst-Controlled Halogenation of Indole C-H Bonds.

ABSTRACT: RebH variants capable of chlorinating substituted indoles ortho-, meta-, and para- to the indole nitrogen were evolved by directly screening for altered selectivity on deuterium-substituted probe substrates using mass spectrometry. This systematic approach allowed for rapid accumulation of beneficial mutations using simple adaptive walks and should prove generally useful for altering and optimizing the selectivity of C-H functionalization catalysts. Analysis of the beneficial mutations showed that structure-guided selection of active site residues for targeted mutagenesis can be complicated either by activity/selectivity tradeoffs that reduce the possibility of detecting such mutations or by epistatic effects that actually eliminate the benefits of a mutation in certain contexts. As a coro

SUBMITTER: Andorfer MC 

PROVIDER: S-EPMC4917012 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature