通过分子对接、MD模拟和实验验证从香梅粉水解物中解锁抗酪氨酸酶和抗黑素肽

Torreya grandis is an economically important forest tree species. As a processing by-product, Torreya grandis meal (TGM) is rich in proteins and bioactive substances. Pepsin was selected as the optimal protease for hydrolyzing TGM. The T3 fraction (< 3 kDa), which showed the strongest tyrosinase-inhibitory activity with an IC₅₀ value of 0.17 mg/mL, was obtained via ultrafiltration. Three novel tyrosinase-inhibitory peptides (LPFASW, WVPVSVE and YEVSL) were obtained through LC-MS/MS, in silico docking and cellular assays. Static molecular docking showed that all three peptides could spontaneously bind to the binuclear copper active cavity of tyrosinase 2Y9X with binding energies below -7 kcal·mol-1, mainly through hydrophobic interactions. The 100 ns molecular dynamics simulations demonstrated that LPFASW and WVPVSVE maintained long-term binding via stable hydrogen-bond networks, whereas the hydrogen bonds of YEVSL tended to dissociate in the early simulation phase, leading to poor binding stability. In vitro tyrosinase inhibition assays gave IC50 values of 232.57, 303.14, and 1009.50 μM for WVPVSVE, LPFASW, and YEVSL, respectively, consistent with MD-predicted binding stability. B16F10 cell experiments confirmed that the cell viability remained above 80% at 0-1200 μg/mL for all peptides, indicating low cytotoxicity. At 300 μg/mL, melanin content was reduced by 13.86%, 19.13% and 37.08% for YEVSL, LPFASW and WVPVSVE, respectively, while intracellular tyrosinase activity was inhibited by 33.20%, 40.64% and 18.50%, respectively. In conclusion, TGM-derived LPFASW, WVPVSVE and YEVSL possess tyrosinase-inhibitory and melanin-suppressing activities and show potential as natural functional food additives and as raw materials for cosmetics targeting pigmentation conditions.