Food-derived peptides with strong calcium-binding and osteogenic activity are promising functional ingredients for osteoporosis intervention. This study aims to explore whether a novel phosvitin phosphopeptide (Glu-Asp-Asp-pSer-pSer, EDDpSpS) with excellent calcium binding ability can promote the osteogenic differentiation of MC3T3-E1 cells and its potential mechanism. Network pharmacology analysis of EDDpSpS regulating osteoblast differentiation identified that the focal adhesion was a key signaling pathway, which is closely related to cell differentiation. Molecular simulations demonstrated that EDDpSpS was strongly bound to the target proteins within the focal adhesion signaling pathway and maintains a stable state. In addition, EDDpSpS had the potential to activate the key target protein Src tyrosine kinase (Src). Results of cell experiments showed that EDDpSpS (> 100 μg/mL) significantly promoted the differentiation and mineralization of MC3T3-E1 cells. Transcriptome analysis verified that the expression of paxillin (PXN), Src, actin beta/gamma 1 (ACTG1), and other osteoblast genes were regulated by EDDpSpS, leading to an up-regulation of the focal adhesion. In this study, the effect and the potential mechanism of EDDpSpS on osteoblast differentiation were clarified. EDDpSpS may be used as an effective dual-effect food ingredient for developing functional foods targeting osteoporosis.
Phosvitin has excellent calcium binding capacity, related to its phosphopeptides. The phosphopeptides may be used as functional ingredients for improving calcium bioavailability, but the calcium-binding mechanism is unclear. In this study, a novel phosvitin phosphorylated pentapeptide (Glu-Asp-Asp-pSer-pSer, EDDpSpS) was selected to prepare an EDDpSpS calcium complex (EDDpSpS-Ca), and the calcium-binding mechanism and bioavailability investigated. The calcium-binding capacity of EDDpSpS was up to 468 ± 152.80 mg/g. Calcium ions prompted the folding of the EDDpSpS structure to form spherical nanoparticles. The calcium binding sites of EDDpSpS involved peptide bonds, carboxyl, amino, and phosphate groups. Molecular forces involved in these interactions were electrostatic in nature. Moreover, EDDpSpS-Ca had excellent bioavailability when compared to CaCO 3 , calcium lactate, and d -calcium gluconate. This study revealed the calcium-binding mechanism of phosvitin phosphopeptide, and suggested that EDDpSpS-Ca has the potential to be a novel, efficient, and promising calcium supplement.
危险品化学品经营许可证（带存储）