Release of Bioactive Peptides from Whey Protein During In Vitro Digestion and Their Effect on CCK Secretion in
Anaís Ignot-Gutiérrez1, Orlando Arellano-Castillo2, Gloricel Serena-Romero2
1Instituto de Neuroetología, Universidad Veracruzana, Av. Dr. Luis Castelazo Ayala s/n, Industrial Ánimas, Xalapa-Enríquez 91193, Veracruz, Mexico.
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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 β-lactoglobulin (β-La) and α-lactalbumin (α-La), enriched in hydrophobic and aromatic residues, including neuropeptide-like sequences containing the Glu-Asn-Ser-Ala-Glu-Pro-Glu (ENSAEPE) motif of β-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.
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