Valorization of Whey into a Bioactive Hydrolysate with ACE-Inhibitory Activity and Food Application Potential.

Bioactive peptides derived from whey proteins have attracted increasing interest as natural dietary modulators of angiotensin-converting enzyme (ACE) activity and potential contributors to cardiovascular health. In this study, bioactive whey hydrolysate (BWH) was obtained by controlled enzymatic hydrolysis with plant-derived proteases and characterized in terms of physicochemical properties, peptide profile, ACE-inhibitory activity, stability, and performance in food matrices. RP-HPLC and LC-MS/MS analysis revealed a complex, reproducible peptidome across production batches, including a conserved set of antihypertensive peptide sequences. BWH exhibited high ACE-inhibitory activity (>90%) with low intra-batch variability, and dose-response analysis yielded an estimated IC50 of 3.20% BWH under the assay conditions. Its bioactivity and peptide profile remained stable during storage at room temperature and under refrigeration, as well as after mild thermal treatment (pasteurization), although partial activity loss was observed after baking at high temperatures. Incorporation trials showed that BWH retained strong ACE-inhibitory functionality when incorporated at 7% into yogurt and orange juice, with minimal effects of pasteurization in the beverage matrix. These findings support the potential use of BWH as a functional ingredient for developing antihypertensive foods and beverages.