Arginine-Containing Peptides Derived from Walnut Protein Against Cognitive and Memory Impairment in Scopolamine-Induced Zebrafish: Design, Release, and Neuroprotection

Abstract

The purpose of this study was to investigate the neuroprotective effect of Arg-containing peptides from walnut storage protein sequences in scopolamine-induced zebrafish and further to validate the potential neuroprotection of Arg-containing peptide enriched walnut hydrolysates prepared by <i>in silico</i> hydrolysis and controlled enzymatic release. Results showed that walnut derived Arg-containing peptides with high abundance and great bioactivity predicted by bioinformatics displayed potent neuroprotection in scopolamine-induced zebrafish, and regulation of neurotransmitter level and antioxidant enzyme activity might be the main target for Arg-containing peptides to exert neuroprotection. Notably, Arg-containing peptides (not free arginine) contributed greater neuroprotection, and the positive charge and cell-penetrating properties also affected their neuroprotection. Subsequently, Arg-containing peptides could be released efficiently from walnut protein following hydrolysis by trypsin, pepsin, papain, and thermolysin (bound arginine content: ranging from 110.43 ± 1.58 to 121.82 ± 1.02 mg/g). Among them, trypsin had excellent potential for releasing Arg-containing peptides <i>in silico</i> hydrolysis, and its hydrolysate was confirmed to have neuroprotective capacity, indicating that the combination of <i>in silico</i> hydrolysis and controlled enzymatic release might be an effective approach to obtain Arg-containing neuroprotective peptides.