The study was the first to evaluate the synergistic interaction of luteolin + quercetin, luteolin + 3-O-methylquercetin, and quercetin + 3-O-methylquercetin mixtures on α-glucosidase and the binding mechanisms were explored using both experimental and theoretical approaches. The results showed that three flavonoid mixtures exhibited a mixed type of inhibition and demonstrated the most potent synergistic effects on α-glucosidase inhibition at 6:4 ratio, with interaction index (γ) of 0.85, 0.78 and 0.73, respectively. The three mixtures had a great influence on α-glucosidase secondary structures. Molecular simulation further demonstrated that three flavonoid mixtures formed hydrophobic interactions and hydrogen bonds with amino acid residues at different sites of α-glucosidase. Collectively, luteolin + quercetin, luteolin + 3-O-methylquercetin and quercetin + 3-O-methylquercetin were found to inhibit α-glucosidase in a synergistic manner and can be potentially used for the development of hypoglycemic food products.
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