Defatted walnut meal (DWM), a main byproduct of walnut oil production, is rich in proteins with a high essential amino acid content. Our research was focused on the functional activities of defatted walnut meal hydrolysate (DWMH). DWMH exhibited relatively strong hydroxyl scavenging and oxygen radical absorbance capacities, protective effect on H2O2-injured PC12 cells compared with GSH and cerebrolysin. Besides, DWMH could combat d-galactose induced learning and memory impairments in the Morris water maze test and in the Dark/light avoidance test of mice. Seventy-seven peptides were identified by UPLC-ESI-MS/MS in the most potent antioxidative fraction of DWMH. WSREEQEREE and ADIYTEEAGR were the peptides that most likely accounted for the suppression of H2O2-induced apoptosis in PC12 cells. The mixture of the commercially available amino acids did not offer any protection on H2O2-injured PC12 cells, suggesting that amino acid sequence rather than amino acid composition was the dominant influencing factor. DWMH could be recommended as natural antioxidants for the development of functional foods targeting memory impairments.