Inhibitory mechanisms of galloylated forms of theaflavins on α-glucosidase

Theaflavins, oxidation product of tea polyphenols, have demonstrated significant inhibitory effects on α-glucosidase, which is beneficial in alleviating hyperglycemia. This study found that the inhibition of four monomers of theaflavins on α-glucosidase was related to the presence of the galloyl moiety (GM), with IC 50 values ranging from TFDG (0.26 mg/mL) < TF3′G (0.33 mg/mL) < TF3G (0.39 mg/mL) ≪ TF (3.26 mg/mL). The multi-spectroscopic analyses revealed that theaflavin monomers changed the microenvironment around aromatic amino acid residues and conformation of α-glucosidase, with the hierarchy being TFDG > TF3′G > TF3G > TF. The binding of theaflavins with α-glucosidase was confirmed by differential scanning calorimetry (DSC), isothermal titration calorimetry (ITC), molecular docking and molecular dynamics simulations analysis. It was confirmed that theaflavins can form stable complexes with α-glucosidase, and that hydrogen bonding and van der Waals forces play important roles in the binding of theaflavins to α-glucosidase. The strongest binding affinity was observed between TFDG and the enzyme's active site, which corresponded with its enzyme activity inhibition ability. The study suggests that GM substitution plays a crucial role in enhancing the binding of theaflavins to α-glucosidase, thereby inducing greater conformational changes and leading to a stronger inhibitory effect on α-glucosidase.