Abstract: In order to further research the inhibiting mechanisms of myricetin on the activity of α-glucosidase, different modern spectral analysis methods were used in this paper, combined with atomic force microscope ( AFM) and molecular docking, to investigate the interactions between myricetin and α-glucosidase.The results indicated that myricetin showed strong inhibition function on the activity of α-glucosidase and the value of IC50 was 0.99 × 10-5 mol/L. The kinetic study revealed that the inhibiting effect between myricetin and α-glucosidase belong to competitive inhibition, and there was a single inhibition site onα-glucosidase, moreover the fluorescence of α-glucosidase was quenched by myricetin through a static quenching mechanism.The molecular docking showed that myricetin could form hydrogen bonds with the amino acids of TYR158, GLN279, GLU277, ASP215 and ASP352 to change the micro-environments of α-glucosidase, which leaded a clustering phenomenon and then had the inhibition activity.