Understanding activity-stability tradeoffs in biocatalysts by enzyme proximity sequencing.

ABSTRACT: Understanding the complex relationships between enzyme sequence, folding stability and catalytic activity is crucial for applications in industry and biomedicine. However, current enzyme assay technologies are limited by an inability to simultaneously resolve both stability and activity phenotypes and to couple these to gene sequences at large scale. Here we present the development of enzyme proximity sequencing, a deep mutational scanning method that leverages peroxidase-mediated radical labeling with single cell fidelity to dissect the effects of thousands of mutations on stability and catalytic activity of oxidoreductase enzymes in a single experiment. We use enzyme proximity sequencing to analyze how 6399 missense mutations influence folding stability and catalytic activity in a D-amin

SUBMITTER: Vanella R 

PROVIDER: S-EPMC10902396 | biostudies-literature | 2024 Feb

REPOSITORIES: biostudies-literature