Copper chelating peptides derived from tilapia (Oreochromis niloticus) skin as tyrosinase inhibitor: Biological evaluation, in silico investigation and in vivo effects.

Binuclear copper ions at the active site determine the catalysis of tyrosinase (TYR) whose activity can be inhibited by copper's chelation with other compounds. In this study, tilapia (Oreochromis niloticus) skin was used to generate TYR-inhibitory peptides after being treated by different enzymes and 4 h-Alcaline protease hydrolysate exhibited the highest TYR inhibition and copper chelation. Immobilized metal affinity chromatography was used for purifying copper chelating peptides, among which PFRMY (IC: 0.43 ± 0.08 mg/mL) and RGFTGM (IC: 1.61 ± 0.04 mg/mL) exhibited the highest TYR-inhibitory capacity and the lowest docking energy. Both two peptides inhibited TYR in a mixed manner and interacted with key residues binding to copper ions within TYR mainly by hydrogen bonds and hydrophobic forces, while PFRMY had a more compact and stable conjugation with TYR. Zebrafish assay revealed that PFRMY reduced not only melanin synthesis but in vivo TYR activity.