Xanthine oxidase is an oxidase that has a molybdopterin structure with substrate inhibition. Here, we show that a single point mutation (Q201) in the Acinetobacter baumannii xanthine oxidase (AbXOD) obtained mutant Q201E (k cat =799.44 s-1, no inhibition) with high enzyme activity and decrease of substrate inhibition in 5 mmol/L high substrate model, and which cause two loops structure change at active center, characterized by complete loss of substrate inhibition without reduction of enzymatic activity. Molecular docking results showed that the change of flexible loop increased the affinity between substrate and enzyme, and the formation of a π-π bond and two hydrogen bonds made the substrate more stable in the active center. Ultimately, Q201E can still maintain better enzyme activity under high purine content (an approximately 7-fold improvement over the wild-type), indicating a broader application prospect in the manufacture of low-purine food.
Keywords: Acinetobacter baumannii; Enzyme engineering; Fish sauce; Point mutation; Substrate inhibition; Xanthine oxidase.
© 2023 The Author(s).