Conformational changes of tyrosinase caused by pentagalloylglucose binding: Implications for inhibitory effect and underlying mechanism.

Tyrosinase is a critical enzyme related to various pigmentation disorders and browning of fruits and vegetables. In this study, a novel inhibitor pentagalloylglucose (PGG) against tyrosinase was prepared from tannic acid with the chemical structure elucidated using HPLC, ESI-MS, H- and C NMR. Its inhibitory effect and the underlying mechanism on tyrosinase were explored by enzyme kinetics, UV-scanning, copper-ion chelation, fluorescence, circular dichroism, fourier transform infrared spectroscopy and molecular docking simulation. Results revealed that the yield of PGG reached 18.0% and the purity was up to 99.09%. PGG was a high-potential inhibitor of tyrosinase with IC values of (15.54 ± 0.56) × 10 and (50.89 ± 3.34) × 10 mol/L for monophenolase and diphenolase, respectively. PGG could disturb the formation of dopachrome and had strong capacity to chelate copper ions. The fluorescence of tyrosinase was efficiently quenched by PGG through a static mechanism. The binding of PGG to tyrosinase was a spontaneous exothermic process that induced unfolding of the tyrosinase structure to expose more buried hydrophobic residues. Docking results implied that PGG interacted with tyrosinase by forming hydrogen bonds with amino acid residues Glu-173, Glu-208, Lys-158, Lys-180, Gln-44 and Gln-159. This study would enhance our understanding of the inhibitory mechanism of PGG on tyrosinase at the molecular level and provide scientific guidance for the application of PGG in food and pharmaceutical industries.