Inhibition characteristics and mechanism of tyrosinase using five citrus flavonoids: A spectroscopic and molecular dynamics simulation study

This work presents a comparative analysis of the tyrosinase inhibitory effects of five citrus flavonoids, namely hesperetin, hesperidin, neohesperidin, naringenin and naringin. Visbile, fluorescence, and fourier transform infrared (FITR) spectroscopies, and molecular dynamic methods were employed to compare the anti-tyrosinase mechanisms of each flavonoid. Hesperetin, neohesperidin, naringenin and naringin exhibited potent inhibitory activities with IC50 values of 0.74 ± 0.05, 2.19 ± 0.03, 7.50 ± 9.82 and 24.94 ± 8.43 μmol/ml, respectively, all of which were higher than that of kojic acid (0.04 ± 0.02 μmol/ml). The enzymatic kinetics results suggested that hesperetin and naringenin were reversible inhibitors on tyrosinase in the mixed-type manner. H-bond and hydrophobic interactions were found to drive the binding of tyrosinase with hesperetin or naringenin, which subsequently changed the FTIR spectroscopy results by decreasing the α-helix ratio and increasing the β-turn, β-sheet and random coil ratio in tyrosinase. Molecular dynamics simulation not only verified some of the experimental results, but also suggested that the binding of hesperetin and naringenin to tyrosinase was spontaneous. The findings of this study indicate that citrus flavonoids are a promising dietary resource for tyrosinase inhibition. PRACTICAL APPLICATIONS: Hesperetin, hesperidin, neohesperidin, naringenin and naringin were typical citrus flavonoids that have anti-obesity, anti-oxidation, anti-inflammation and anti-diabetes activities. Current study suggested that hesperetin and naringenin were effective reversible inhibitors on tyrosinase in the mixed-type manner. Hesperetin and naringenin might serve as nutritional and chemical agents for regulating the tyrosinase activity to control melanin level in vivo.