Purification, Identification and Molecular Docking of Novel Antioxidant Peptides from Walnut (Juglans regia L.) Protein Hydrolysates

Abstract

Walnut protein isolate (WPI) was hydrolyzed using Alcalase for 0, 30, 60, 90, 120 and 150 min to investigate the effect of different hydrolysis times on the structure and antioxidant properties of walnut proteins. The identified peptides HADMVFY, NHCQYYL, NLFHKRP and PSYQPTP were used to investigate the structure-activity relationship by using LC-MS/MS and molecular docking. The kinetic equations DH = 3.72ln [1 + (6.68 E0/S0 + 0.08) t] were developed and validated to explore the mechanism of WIP hydrolysis by Alcalase. Structural characteristics showed that the UV fluorescence intensity and endogenous fluorescence intensity of the hydrolysates were significantly higher than those of the control. FTIR results suggested that the secondary structure gradually shifted from an ordered to a disordered structure. Enzymatic hydrolysis containing much smaller molecule peptides than WPI was observed by molecular weight distribution. In vitro, an antioxidant test indicated that Alcalase protease hydrolysis at 120 min showed more potent antioxidant activity than hydrolysates at other hydrolysis times. In addition, four new antioxidant peptides were identified by LC-MS/MS. Molecular docking indicated that these peptides could interact with ABTS through interactions such as hydrogen bonding and hydrophobic interactions. Thus, WPI hydrolysates could be used as potential antioxidants in the food and pharmaceutical industries.

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