Complete computational design of high-efficiency Kemp elimination enzymes.

ABSTRACT: Until now, computationally designed enzymes exhibited low catalytic rates^1-5 and required intensive experimental optimization to reach activity levels observed in comparable natural enzymes^5-9. These results exposed limitations in design methodology and suggested critical gaps in our understanding of the fundamentals of biocatalysis^10,11. We present a fully computational workflow for designing efficient enzymes in TIM-barrel folds using backbone fragments from natural proteins and without requiring optimization by mutant-library screening. Three Kemp eliminase designs exhibit efficiencies greater than 2,000 M^-1 s^-1. The most efficient shows more than 140 mutations from any natural protein, including a novel active site. It exhibits hig

SUBMITTER: Listov D 

PROVIDER: S-EPMC12310539 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature