Efficient Molecular Evolution to Generate Enantioselective Enzymes Using a Dual-channel Microfluidic Droplet Screening Platform

Overview

Journal Nat Commun

Specialty Biology

Date 2018 Mar 14

PMID 29531246

Citations 34

Authors

Fuqiang Ma

Meng Ting Chung

Yuan Yao

Robert Nidetz

Lap Man Lee

Allen P Liu

Yan Feng

Katsuo Kurabayashi

Guang-Yu Yang

Affiliations

Soon will be listed here.

Abstract

Directed evolution has long been a key strategy to generate enzymes with desired properties like high selectivity, but experimental barriers and analytical costs of screening enormous mutant libraries have limited such efforts. Here, we describe an ultrahigh-throughput dual-channel microfluidic droplet screening system that can be used to screen up to ~10 enzyme variants per day. As an example case, we use the system to engineer the enantioselectivity of an esterase to preferentially produce desired enantiomers of profens, an important class of anti-inflammatory drugs. Using two types of screening working modes over the course of five rounds of directed evolution, we identify (from among 5 million mutants) a variant with 700-fold improved enantioselectivity for the desired (S)-profens. We thus demonstrate that this screening platform can be used to rapidly generate enzymes with desired enzymatic properties like enantiospecificity, chemospecificity, and regiospecificity.

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