Ultrahigh-Throughput Directed Evolution of Polymer-Degrading Enzymes Using Yeast Display.

ABSTRACT: Enzymes that degrade synthetic polymers have attracted intense interest for eco-friendly plastic recycling. However, because enzymes did not evolve for the cleavage of abiotic polymers, directed evolution strategies are needed to enhance activity for plastic degradation. Previous directed evolution efforts relied on polymer degradation assays that were limited to screening ∼10⁴ mutants. Here, we report a high-throughput yeast surface display platform to rapidly evaluate >10⁷ enzyme mutants for increased activity in cleaving synthetic polymers. In this platform, individual yeast cells display distinct mutants, and enzyme activity is detected by a change in fluorescence upon the cleavage of a synthetic probe resembling a polymer of interest. Highly active mutants are is

SUBMITTER: Cribari MA 

PROVIDER: S-EPMC11058326 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature