Mutant β-Glucosidases: Structural Insights for Biocatalytic Applications

Abstract: β-glucosidase enzymes play a pivotal role in the hydrolysis of glycosidic bonds, with bio-catalytic applications in glycoside synthesis, lignocellulosic biomass degradation, and the processing of glycosides. This review explores the engineering of β-glucosidase mutants to enhance their cata-lytic efficiency, substrate specificity, and stability for biocatalytic applications. It also considers hy-drolytic applications of glucosidases, such as lignocellulosic biomass valorisation, as well as syn-thetic applications via reverse hydrolysis, including the production of alkyl glycosides. Additionally, it examines rational design-led mutagenesis and directed evolution strategies to tailor enzyme activity, along with structural insights gained from molecular dynamics simulations and mo-lecular docking studies. These structural insights provide an understanding of the active-site archi-tecture, catalytic mechanism, and enzyme-substrate interactions, which are useful for the rational design of improved glucosidase variants. This review underscores the synergistic role of mutagenesis studies and structural analysis in optimizing glucosidases for efficient and sustainable biocatalytic applications, paving the way for innovative applications in green chemistry.