Structure identification of walnut peptides and evaluation of cellular antioxidant activity

Walnut peptides are widely accepted as nutraceuticals with multiple health benefits. However, information about the structure and biological activity of walnut peptides is limited. In this work, walnut protein hydrolysate (WPH) was prepared by enzymatic hydrolysis. The total and free amino acids compositions in WPH were determined. The peptides sequence of WPH was analyzed by UPLC-MS/MS, and forty eight bioactive peptides were identified. Antioxidant activities were detected for WPH in both extracellular and intercellular assays. QGRPWG, PSRADIY and AYNIPVNIAR were the peptides that accounted for the antioxidant activity of WPH. Among them, QGRPWG showed the highest ORAC value (2801 µmol TE/g), and AYNIPVNIAR possessed the strongest protective effect against H₂O₂-induced oxidative stress in PC-12 cells in a dose-dependent manner. The results can aid in the recognition of peptides from walnut as nutraceuticals.