Enzymatic hydrolysis of cassava leaf proteins unlocks antioxidant and therapeutic peptides: Alcalase-derived bioactives target oxidative stress pathways

Cassava leaves, often discarded as agricultural byproduct, are a nutrient-dense yet underutilized protein source, containing 38 % proteins by dried weight. This study investigates the potential of cassava leaf proteins as a sustainable alternative to conventional protein sources through enzymatic hydrolysis. Bioactive peptides extracted during hydrolysis significantly improve the protein's nutritional profile and exhibited diverse biological activities. Analysis revealed a notable increase in essential amino acids, comprising ∼40 % of total amino acids. Using LC-MS/MS, 36 distinct peptide sequences were identified. Bioinformatics and network pharmacology analyses revealed that cassava leaf peptides possess antioxidant, anti-inflammatory, immunomodulatory, anticancer, and neuroprotective properties. Notably, the peptide sequences HPDGF and SEGGF exhibited strong binding affinity to key targets (AKT1, CASP3, CCND1, and IL1B), suggesting therapeutic potential for oxidative stress-related pathologies. These findings underscore the potential of cassava leaf proteins as a natural, sustainable source of bioactive peptides for health applications, notably as antioxidants.