Thermoadaptation in an Ancestral Diterpene Cyclase by Altered Loop Stability.

ABSTRACT: Thermostability is the key to maintain the structural integrity and catalytic activity of enzymes in industrial biotechnological processes, such as terpene cyclase-mediated generation of medicines, chiral synthons, and fine chemicals. However, affording a large increase in the thermostability of enzymes through site-directed protein engineering techniques can constitute a challenge. In this paper, we used ancestral sequence reconstruction to create a hyperstable variant of the ent-copalyl diphosphate synthase PtmT2, a terpene cyclase involved in the assembly of antibiotics. Molecular dynamics simulations on the μs timescale were performed to shed light on possible molecular mechanisms contributing to activity at an elevated temperature and the large 40 °C increase in melting tempera

SUBMITTER: Hueting DA 

PROVIDER: S-EPMC9169049 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature