Changes in Active Site Loop Conformation Relate to the Transition toward a Novel Enzymatic Activity.

ABSTRACT: Enzymatic promiscuity, the ability of enzymes to catalyze multiple, distinct chemical reactions, has been well documented and is hypothesized to be a major driver of the emergence of new enzymatic functions. Yet, the molecular mechanisms involved in the transition from one activity to another remain debated and elusive. Here, we evaluated the redesign of the active site binding cleft of lactonase SsoPox using structure-based design and combinatorial libraries. We created variants with largely improved catalytic abilities against phosphotriesters, the best ones being >1000-fold better compared to the wild-type enzyme. The observed shifts in activity specificity are large, and some variants completely lost their initial activity. The selected combinations of mutations have considerabl

SUBMITTER: Jacquet P 

PROVIDER: S-EPMC11134384 | biostudies-literature | 2024 May

REPOSITORIES: biostudies-literature