Release of Bioactive Peptides from Whey Protein During In Vitro Digestion and Their Effect on CCK Secretion in Enteroendocrine Cells: An In Silico and In Vitro Approach Academic Article in Scopus uri icon

abstract

  • During gastrointestinal digestion, dietary proteins are hydrolyzed into peptides and free amino acids that modulate enteroendocrine function and satiety-related hormone secretion along the gut¿brain axis, thereby contributing to obesity prevention. We investigated whey protein concentrate (WPC) as a source of bioactive peptides and evaluated the effects of its digests on cholecystokinin (CCK) secretion in STC-1 enteroendocrine cells by integrating the standardized INFOGEST in vitro digestion protocol, peptidomics (LC¿MS/MS), and in silico bioactivity prediction. In STC-1 cells, the <3 kDa intestinal peptide fraction exhibited the strongest CCK stimulation, positioning these low-molecular-weight peptides as promising bioactive components for satiety modulation and metabolic health applications. Peptidomic analysis of this fraction identified short sequences derived primarily from ß-lactoglobulin (ß-La) and ¿-lactalbumin (¿-La), enriched in hydrophobic and aromatic residues, including neuropeptide-like sequences containing the Glu¿Asn¿Ser¿Ala¿Glu¿Pro¿Glu (ENSAEPE) motif of ß-La f(108¿114). In silico bioactivity profiling with MultiPep predicted antihypertensive, angiotensin-converting enzyme (ACE)¿inhibitory, antidiabetic, dipeptidyl peptidase-IV (DPP-IV)¿inhibitory, antioxidant, antibacterial, and neuropeptide-like activities. Overall, digestion of WPC released low-molecular-weight peptides and amino acids that enhanced CCK secretion in vitro; these findings support their potential use in nutritional strategies to enhance satiety, modulate appetite and energy intake, and improving cardiometabolic health. © 2026 by the authors.

publication date

  • January 1, 2026