Enabling high-throughput enzyme discovery and engineering with a low-cost, robot-assisted pipeline.

ABSTRACT: As genomic databases expand and artificial intelligence tools advance, there is a growing demand for efficient characterization of large numbers of proteins. To this end, here we describe a generalizable pipeline for high-throughput protein purification using small-scale expression in E. coli and an affordable liquid-handling robot. This low-cost platform enables the purification of 96 proteins in parallel with minimal waste and is scalable for processing hundreds of proteins weekly per user. We demonstrate the performance of this method with the expression and purification of the leading poly(ethylene terephthalate) hydrolases reported in the literature. Replicate experiments demonstrated reproducibility and enzyme purity and yields (up to 400 µg) sufficient for comprehensive analyses of

SUBMITTER: Norton-Baker B 

PROVIDER: S-EPMC11196671 | biostudies-literature | 2024 Jun

REPOSITORIES: biostudies-literature