Comprehensive characterization of bioactive peptides from Buffalo (Bubalus bubalis) colostrum and milk fat globule membrane proteins

•Fat globule membrane proteins from buffalo milk and colostrum•Characterization of peptides (< 3 Da) after in vitro gastrointestinal simulation digestion using nano-LC MS/MS analysis•Functional cataloguing of fat globule membrane proteins using peptide tags•Subcellular localization and establishment of protein interacting networks among fat globule membrane proteins•Cytoprotective role of peptides against oxidative stress

Milk fat is dispersed in milk as small spherical globules stabilized in the form of emulsion by its surrounding membrane, often referred to as fat globule membrane (FGM). Buffalo, a major milking mammal of Asia and second most milking mammal across the globe presents physicochemical features different from that of other ruminant species containing higher content of lipids and proteins. The present study describes characterization of FGM proteins isolated from both buffalo milk and colostrum. A detailed proteomic analysis of peptides generated by in vitro gastrointestinal simulation digestion of buffalo milk and colostrum FGM fractions was performed by nLC-ESI MS/MS. The peptide based clustering of FGM proteins unravelled association of membrane proteins in fat transport, enzymatic activity, general transport, defence, cell signalling, membrane/protein trafficking protein synthesis/binding/folding including unknown functions. Gene annotation, STRING and YLoc analyses provided putative insights into major secretory pathways in milk and colostrum FGM peptides, interactive protein networks including their sub cellular localization. The peptides of milk and colostrum FGM offered cellular protection as powerful antioxidants indicated their promising perspectives in commercial formulations and nutraceuticals.

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