The Potential for Bioactive Peptide Production in a Fermented Dairy Beverage Based on Chickpea Water Extract Using Proteolytic Lactic Acid Bacteria

A chickpea-based milk beverage containing both plant and animal proteins represents an excellent substrate for the production of biologically active peptides through fermentation. Fermentation by lactic acid bacteria (LAB) increases its nutritional value compared to the unfermented beverage while improving the digestibility and bioavailability of essential nutrients via proteolytic enzyme activity. This study investigated the production of bioactive peptides in fermented chickpea water extract using ten bacterial strains isolated from plant and animal sources. The proteolytic activity of each strain was quantified using the trinitrobenzene sulfonic acid (TNBS) method, and the presence of proteolytic genes was confirmed via agarose gel electrophoresis. Peptides released during fermentation were identified through two-dimensional electrophoresis, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and tandem mass spectrometry. To predict the potential biological activities of the studied peptide sequences, a series of in silico analyses were performed using specialized bioinformatics tools. The identified peptides were predicted to exhibit antioxidant, antihypertensive, anticancer, antibacterial, antifungal, antituberculosis, and angiotensin-converting enzyme (ACE) inhibitory activities. Based on the results, L. fermentum SB-2 and L. sakei SD-8, were selected as promising candidates for bioactive peptide production in a chickpea water extract-based milk beverage and were subsequently applied in the beverage prototype.