Tyrosinase inhibitors have important applications in the cosmetics, medical and food industries due to they can effectively inhibit the synthesis of melanin. In this study, tilapia scale polypeptides were used as raw materials, and high-purity polypeptides with metal copper ions chelating ability were obtained by enzymatic hydrolysis, column chromatography, and EDTA elution. In vitro cell model analysis showed that the fish squamous peptides could strongly inhibit the activity of tyrosinase. When the sample concentration was 5 mg·mL-1, its inhibition rate of tyrosinase reached to 59.73%, which had a better inhibition of enzyme activity compared with the positive control of the same concentration. The comprehensive results showed that the fish scale polypeptide with metal copper ions chelating ability could be a strong tyrosinase inhibitor, and might be used to prevent food browning in food-related fields, and could also be used for skin whitening in the fields of medicine and cosmetics.