High throughput screening with SAMDI mass spectrometry for directed evolution

Abstract

Advances in directed evolution have led to an exploration of new and important chemical transformations; however, many of these efforts still rely on the use of low-throughput chromatography-based screening methods. We present a high-throughput strategy for screening libraries of enzyme variants for improved activity. Unpurified reaction products are immobilized to a self-assembled monolayer and analyzed by mass spectrometry, allowing for direct evaluation of thousands of variants in under an hour. The method was demonstrated with libraries of randomly mutated cytochrome P411 variants to identify improved catalysts for C–H alkylation. The technique may be tailored to evolve enzymatic activity for a variety of transformations where higher throughput is needed.

ICB Affiliated Authors

Authors
Adam J. Pluchinsky, Daniel J. Wackelin, Xiongyi Huang, Frances H. Arnold, Milan Mrksich
Date
Type
Peer-Reviewed Article
Journal
Journal of the American Chemical Society
Volume
142
Number
47
Pages
19804−19808
Emblems