Tyrosinase is the main rate-limiting enzyme of melanogenesis, and inhibition of tyrosinase activity is one of the effective ways to reduce melanogenesis and thus achieve a whitening effect.In this experiment, the inhibitory effects and mechanisms of cyanidin-3-O-glucoside (C3G) and cyanidin (Cy) on tyrosinase were comprehensively investigated using enzyme inhibition kinetic analysis, ultraviolet spectroscopy, infrared spectroscopy, fluorescence spectroscopy, circular dichroism spectroscopy, and molecular docking simulation analysis while using mushroom tyrosinase as the target.Results showed that the inhibitory activity of C3G (IC50 =124.8 μmol/L) and Cy (IC50 =278.1 μmol/L) on tyrosinase were better than that of the positive control kojic acid (IC50 =536.4 μmol/L).The inhibitory effect of C3G and Cy on tyrosinase was reversible, and the inhibitory effect of C3G on tyrosinase was competitive inhibition, while the inhibitory effect of Cy on tyrosinase was mixed inhibition.Results of ultraviolet, Fourier transform infrared spectroscopy, and circular dichroism spectra showed that the binding of C3G and Cy to tyrosinase resulted in the conformation change of tyrosinase, which led to the inhibition of the enzyme activity.Fluorescence quenching results showed that C3G and Cy combined with tyrosinase changed the microenvironment of Tyr and Trp residues, resulting in static quenching.Synchronous fluorescence results showed that the combination of C3G and Cy with tyrosinase enhanced the microenvironmental polarity of Tyr and Trp residues and reduced the hydrophobic ability.Three-dimensional fluorescence spectra showed that both C3G and Cy could reduce the intensity of spectral characteristic peaks of tyrosinase.Results of the multispectral analysis showed that the inhibitory effect of C3G on tyrosinase was stronger than that of Cy.Molecular docking simulation results showed that C3G formed six hydrogen bonds with amino acid residues HIS178, ASN174, GLN41, and GLN44 of tyrosinase, and π-π interaction was generated with amino acid residues GLU173 and LYS180.Cy bound to tyrosinase and formed four hydrogen bonds with amino acid residues ASN174, GLU173, GLN41, and GLN44, resulting in π-π interaction with amino acid residues GLU173.Molecular docking results further demonstrated that C3G bound tightly to residues in the active cavity of tyrosinase and had a higher affinity for tyrosinase compared to Cy.The conclusions of the inhibitory enzyme activity experiment and multispectral experiment were further supplemented.This study provides a reference for further research and development of polyphenols such as C3G and Cy in the field of tyrosinase inhibitors and acts as scientific support for the development of natural high-efficiency tyrosine inhibitors for cosmetic whitening additives.
ZHANG Chunzi
,
MA Xiaoyan
,
ABLIZ·Abdurahman
,
PANG Tuxia
,
MA Xiaoli
. Inhibition effects and mechanisms of cyanidin-3-O-glicoside and cyanidin on tyrosinase[J]. Food and Fermentation Industries, 2025
, 51(10)
: 224
-232
.
DOI: 10.13995/j.cnki.11-1802/ts.039838
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