Effect of Moringa oleifera Seeds Rennet on the Quality of Mozzarella Cheese and Proteomic Identification and Molecular Docking Analysis of Fermentation-Derived Peptides

Abstract

This study aimed to characterize bioactive water-soluble peptides (WSPs) in Moringa oleifera seeds rennet-derived cheese (MSRC) and compare them with those in calf (CRC), papain (PRC), and microbial rennet cheese (MRC). Proteolysis of MSRC was enhanced, as indicated by its higher hydrolysis indices compared to those of CRC (P < 0.05). The <3 kDa WSP fraction from MSRC had stronger antioxidant activity than those from PRC and MRC (P < 0.05). The fraction also had higher α-glucosidase inhibitory activity (66.19%) and xanthine oxidase inhibitory activity (56.07%) than CRC (P < 0.05). LC–MS/MS identified 2521 peptides in MSRC. 1196 peptides were shared among all cheeses, and 157 were unique to MSRC. Among these, nine characteristic active peptides (e.g., AWPQYLK and FPGPIP) are predicted to exert effects mainly by binding to receptor proteins via hydrogen bonding and hydrophobic interactions. These results highlight MSRC’s potential as a viable alternative to rennet.