Identification of antidiabetic peptides from broad bean protein: Sequencing using LC-MS-QTOF and in-vitro confirmative studies

The global rise in diabetes, driven by sedentary lifestyles, poor dietary habits, obesity, and aging populations, necessitates exploring safer and more affordable treatment options beyond synthetic drugs, which often have adverse side effects and are financially inaccessible in developing nations. This study investigates the potential of bioactive peptides derived from broad bean (faba bean) proteins as natural antidiabetic agents. Broad beans were selected because of their high protein content (22%-36%) and favorable nutritional profile. Broad bean protein hydrolysates were produced using alcalase enzyme under varying hydrolysis conditions to optimize the generation of peptides with alpha-amylase dipeptidyl peptidase-IV (DPP-IV) and alpha-glucosidase inhibitory activities. The hydrolysates exhibited significant degrees of hydrolysis, with the 4-h treatment showing the highest degree, indicating efficient peptide bond cleavage. Furthermore, fractionation of the hydrolysates into different sizes using Amicon Ultra-4 centrifugal filter membrane yielded peptide fractions (BBP10, BBP30, BBP100), which demonstrated notable inhibitory effects on alpha-amylase, alpha-glucosidase, and DPP-IV enzyme, with BBP30 showing the most potent activity. LC-MS-QTOF identified 8 peptides in BBP30, which were further analyzed for bioactivity using in silico analysis. The identified peptides such as ADFF, FPYL, LIPPGM, HGAPILC, GPCSTAGF, FPMFFEG, PEAHFQFF, and PDPPPQVGINC exhibited multifunctional inhibitory activities and favorable physicochemical properties, signifying their effectiveness in lifestyle disease management. These findings highlight the potential of broad bean-derived peptides as natural, bioactive compounds for developing functional foods and nutraceuticals to manage diabetes, providing a promising alternative to conventional synthetic drugs.