A High-Throughput Screening Platform for Engineering Poly(ethylene Terephthalate) Hydrolases.

ABSTRACT: The ability of enzymes to hydrolyze the ubiquitous polyester, poly(ethylene terephthalate) (PET), has enabled the potential for bioindustrial recycling of this waste plastic. To date, many of these PET hydrolases have been engineered for improved catalytic activity and stability, but current screening methods have limitations in screening large libraries, including under high-temperature conditions. Here, we developed a platform that can simultaneously interrogate PET hydrolase libraries of 10⁴-10⁵ variants (per round) for protein solubility, thermostability, and activity via paired, plate-based split green fluorescent protein and model substrate screens. We then applied this platform to improve the performance of a benchmark PET hydrolase, leaf-branch compost cutinas

SUBMITTER: Groseclose TM 

PROVIDER: S-EPMC11459431 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature