The peptide components of defatted walnut ( Juglans regia L.) meal hydrolysate (DWMH) remain unclear, hindering the investigation of biological mechanisms and exploitation of bioactive peptides. The present study aims to identify the peptide composition of DWMH, followed by to evaluate in vitro antioxidant effects of selected peptides and investigate mechanisms of antioxidative effect. First, more than 1 000 peptides were identified by de novo sequencing in DWMH. Subsequently, a scoring method was established to select promising bioactive peptides by structure based screening. Eight brand new peptides were selected due to their highest scores in two different batches of DWMH. All of them showed potent in vitro antioxidant effects on H2O2-injured nerve cells. Four of them even possessed significantly stronger effects than DWMH, making the selected bioactive peptides useful for further research as new bioactive entities. Two mechanisms of hydroxyl radical scavenging and ROS reduction were involved in their antioxidative effects at different degrees. The results showed peptides possessing similar capacity of hydroxyl radical scavenging or ROS reduction may have significantly different in vitro antioxidative effects. Therefore, comprehensive consideration of different antioxidative mechanisms were suggested in selecting antioxidative peptides from DWMH.
Keywords: ROS reduction; antioxidant peptides; de novo sequencing; defatted walnut meal hydrolysate; hydroxyl radical scavenging.