Antioxidant peptides from lupin hydrolysates identified by integrated peptidomic analysis, molecular docking and in vitro assays

Oxidative stress, associated with cardiovascular, neurodegenerative, and cancer-related disorders, may be mitigated by dietary intake of plant-derived antioxidants. In this study, lupin protein hydrolysates were generated using Proteinase K, Papain, and Biopeptidase. HLPI-3 showed the highest antioxidant activity in vitro. A total of 980 peptides with significant differences among enzymatic treatments were identified. Additionally, correlation analysis with antioxidant activity pointed out 36 peptides mainly from Hsp90, conglutin beta, and legumin-like proteins as the most likely contributors to antioxidant activity. Within these, bioinformatic predictions of peptide antioxidant capacity indicated 29 ranked highly (Pre-Score > 0.99) and seven top candidates were prioritized. Docking analysis against myeloperoxidase highlighted four peptides (GQNPLFPR, FLVPPPQS, FLVPPPQSQL, EKLPGGCGGQDH) with strong binding affinities (−8.5 to −9.9 kcal·mol −1 ). These results demonstrate that integrating experimental screening with bioinformatic analysis provided a robust strategy for identifying promising antioxidant peptides from lupin proteins. • Lupin protein hydrolysates are a rich source of antioxidant peptides. • Peptide identification and quantification of lupin hydrolysates using SWATH-MS. • Correlation analysis identified 36 candidate antioxidant peptides. • Bioinformatics prioritized seven peptides with high predicted antioxidant activity. • Molecular docking showed four peptides strongly bind Myeloperoxidase.