Effect of enzymatic hydrolysis of pulse protein macromolecules to tailor structure for enhanced nutraceutical properties

The rising emphasis on health and well-being has driven interest in nutraceuticals, particularly plant-based proteins, due to their combined nutritional and therapeutic benefits. Pulses, with their substantial protein content, hold promise for nutraceutical applications. However, their protein digestibility is limited by anti-nutritional factors. For this purpose, enzymatic modification presents a possible solution to enhance their functionality. Therefore, the objective of the study was to isolate the proteins from fava, lentil, and mung beans, and their hydrolysates were produced by subjecting them to enzymatic treatment using bromelain and thermolysin. The produced protein hydrolysates were evaluated for structural, physico-chemical and nutraceutical properties. Experimental data revealed that thermolysin exhibited higher hydrolysis rates than bromelain. SDS-PAGE analysis displayed extensive breakdown of proteins, forming peptides of varied sizes. SEM analysis revealed altered surface morphology. Circular dichroism (CD) spectroscopy highlighted changes in secondary protein structure post enzymatic treatment. Fourier Transform Infrared (FTIR) spectroscopy showed modifications in functional groups, confirming enzymatic hydrolysis-induced changes in secondary structure. Furthermore, hydrolysis significantly enhanced DPP-IV, α-glucosidase and α-amylase inhibition activities, promising for glycemic control. Pancreatic lipase and cholesterol esterase inhibition activities were also found to increase after hydrolysis, indicating their potential as functional food ingredients.