Rigidifying a De Novo Enzyme Increases Activity and Induces a Negative Activation Heat Capacity.

ABSTRACT: Conformational sampling profoundly impacts the overall activity and temperature dependence of enzymes. Peroxidases have emerged as versatile platforms for high-value biocatalysis owing to their broad palette of potential biotransformations. Here, we explore the role of conformational sampling in mediating activity in the de novo peroxidase C45. We demonstrate that 2,2,2-triflouoroethanol (TFE) affects the equilibrium of enzyme conformational states, tending toward a more globally rigid structure. This is correlated with increases in both stability and activity. Notably, these effects are concomitant with the emergence of curvature in the temperature-activity profile, trading off activity gains at ambient temperature with losses at high temperatures. We apply macromolecular rate theo

SUBMITTER: Hindson SA 

PROVIDER: S-EPMC8453482 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature