Identification, characterization and in vitro activity of hypoglycemic peptides in whey hydrolysates from rubing cheese by-product.

The by-product of Chinese rubing cheese is rich in whey protein. Whey hydrolysates exhibit good hypoglycemic activity, but which specific peptide components are responsible for this effect have not yet been investigated. Herein, the α-glucosidase inhibitory activity of the ultrafiltered fraction (<3 kDa) of rubing cheese whey hydrolysates was evaluated with the inhibition rate of 37.89 %. In addition, peptide identification was conducted using LC-MS/MS, and three peptides YPVEPF, VPYPQ, and LPYPY were identified. Among these, YPVEPF had higher α-glucosidase inhibitory activity (IC50 = 3.52 mg/mL) and interacted with α-glucosidase via hydrogen bonding and hydrophobic forces. YPVEPF was characterized as an amphipathic peptide rich in antiparallel (50.50 %) and random coil (35.20 %) structures, as well as showed good tolerance to gastrointestinal digestion and incubation under the temperature range of 20-80 °C. Notably, YPVEPF activity increased in the presence of Al3+ and Fe3+, as well as within the pH range of 2.0-6.0. Furthermore, YPVEPF had negligible hemolytic activity at a concentration of 1.0 mg/mL, no toxicity at concentrations below 0.5 mg/mL, and significantly promoted glucose consumption in HepG2 cells (p < 0.0001). Collectively, these findings indicate the potential of YPVEPF to be used as a novel hypoglycemic peptide in functional foods.

Characterization of the structure, stability, and activity of hypoglycemic peptides from Moringa oleifera seed protein hydrolysates.

Moringa oleifera seed protein hydrolysates exhibit good hypoglycemic activity, but their specific peptide components have not yet been characterized. Here, we identified the ultrafiltration peptide components (<3 kDa) of M. oleifera seed protein hydrolysates. A highly active α-glucosidase inhibitory peptide with an IC50 value of 109.65 µM (MoHpP-2) with the amino acid sequence KETTTIVR was identified. We characterized its structural properties, stability, and hypoglycemic activity. MoHpP-2 was found to be an amphipathic peptide with a ß-turn structure, and the hemolysis of red blood cells was not observed when its concentration was lower than 2 mg mL-1. MoHpP-2 was stable under weakly acidic conditions, at temperatures lower than 60 °C, and at high ion concentrations. Western blotting revealed that MoHpP-2 affected the PI3K and AMPK pathways of HepG2 cells. Molecular docking revealed that MoHpP-2 interacted with α-glucosidase through hydrogen bonding and hydrophobic forces. Thus, MoHpP-2 from M. oleifera seeds could be used to make hypoglycemic functional foods.