Shuffling the Neutral Drift of Unspecific Peroxygenase in Saccharomyces cerevisiae.

ABSTRACT: Unspecific peroxygenase (UPO) is a highly promiscuous biocatalyst, and its selective mono(per)oxygenase activity makes it useful for many synthetic chemistry applications. Among the broad repertory of library creation methods for directed enzyme evolution, genetic drift allows neutral mutations to be accumulated gradually within a polymorphic network of variants. In this study, we conducted a campaign of genetic drift with UPO in Saccharomyces cerevisiae, so that neutral mutations were simply added and recombined in vivo With low mutational loading and an activity threshold of 45% of the parent's native function, mutant libraries enriched in folded active UPO variants were generated. After only eight rounds of genetic drift and DNA shuffling, we identified an ensemble of 25 n

SUBMITTER: Martin-Diaz J 

PROVIDER: S-EPMC6052263 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature