An Inferred Ancestral CotA Laccase with Improved Expression and Kinetic Efficiency.

ABSTRACT: Laccases are widely used in industrial production due to their broad substrate availability and environmentally friendly nature. However, the pursuit of laccases with superior stability and increased heterogeneous expression to meet industry demands appears to be an ongoing challenge. To address this challenge, we resurrected five ancestral sequences of laccase BsCotA and their homologues. All five variants were successfully expressed in soluble and functional forms with improved expression levels in Escherichia coli. Among the five variants, three exhibited higher catalytic rates, thermal stabilities, and acidic stabilities. Notably, AncCotA2, the best-performing variant, displayed a k_cat/K_M of 7.5 × 10⁵ M^-1·s^-1,

SUBMITTER: Lei L 

PROVIDER: S-EPMC10341886 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature