RESUMO
The insight into the binding mechanism of hesperetin, an aglycone flavanone, with human salivary α-amylase (HSAA), simulated under physiological salivary condition, was explored using various spectroscopic approaches and in silico method. Hesperetin effectively quenched the intrinsic fluorescence of HSAA and the quenching was mixed quenching mechanism. The interaction perturbed the HSAA intrinsic fluorophore microenvironment and the enzyme global surface hydrophobicity. The negative values of ΔG for thermodynamic parameters and in silico study revealed the spontaneity of HSAA-hesperetin complex while the positive values of enthalpy change (ΔH) and entropy change (ΔS) showed noticeable involvement of hydrophobic bonding in the stabilization of the complex. Hesperetin was a mixed inhibitor for HSAA with a KI of 44.60 ± 1.63 µM and having apparent inhibition coefficient (α) of 0.26. Macromolecular crowding, given rise to microviscosity and anomalous diffusion, regulated the interaction. Sodium ion (Na+) created high ionic strength, also, modulated the interaction. The in silico study proposed the preferential binding of hesperetin at the active cleft domain of HSAA with the least energy of -8.0 kcal/mol. This work gives a novel insight on the potentials of hesperetin as a future prospective medicinal candidate in the management of postprandial hyperglycemic condition.Communicated by Ramaswamy H. Sarma.