Computational insights for the hydride transfer and distinctive roles of key residues in cholesterol oxidase

摘要: Cholesterol oxidase (ChOx), a member of the glucose-methanol-choline (GMC) family, catalyzes oxidation substrate via hydride transfer mechanism and concomitant reduction FAD cofactor. Unlike other GMC enzymes, conserved His447 is not catalytic base that deprotonates in ChOx. Our QM/MM MD simulations indicate Glu361 residue acts as facilitating from to We find two rationally chosen point mutations (His447Gln His447Asn) cause notable decreases activity. The binding free energy calculations show residues are important binding. also performed high-level double-hybrid density functional theory using small model systems, which support results. work provides basis for unraveling enzymes histidine does act base.