Efficient molecular evolution to generate enantioselective enzymes using a dual-channel microfluidic droplet screening platform.

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 ~10⁷ 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)-

SUBMITTER: Ma F 

PROVIDER: S-EPMC5847605 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature