Mechanistic Study of Protein Interaction with Natto Inhibitory Peptides Targeting Xanthine Oxidase: Insights from Machine Learning and Molecular Dynamics Simulations

Bioactive peptides from food sources offer a safe and biocompatible approach to enzyme inhibition, with potential applications in managing metabolic disorders such as hyperuricemia and gout, conditions linked to excessive xanthine oxidase activity. Using a machine learning-based screening approach inspired by the bioactivity of natto, two peptides, ECFK and FECK, were identified from the <i>Bacillus subtilis</i> proteome and validated as xanthine oxidase inhibitors with IC₅₀ values of 37.36 and 71.57 mM, respectively. Further experiments confirmed their safety through cytotoxicity assays, and electronic tongue analysis demonstrated their mild sensory properties, supporting their edibility. Molecular dynamics simulations revealed that these peptides stabilize critical enzyme regions, with ECFK showing a higher dissociation energy barrier (52.08 kcal/mol) than FECK (46.39 kcal/mol), indicating strong, stable interactions. This study highlights food-derived peptides as safe and natural inhibitors of xanthine oxidase, offering promising therapeutic potential for metabolic disorder management.