Phosvitin phosphopeptide preparation using immobilised trypsin and enhancing calcium absorption in growing rats

Abstract

We demonstrate a new, efficient method for the preparation of phosvitin phosphopeptides using immobilised-trypsin enzymolysis technology. Immobilised trypsin was prepared using a covalent binding method, and then was added to degrade egg yolk phosvitin for the production of phosphopeptides. In our results, the prepared immobilised trypsin demonstrated a higher hydrolysing activity toward phosvitin than free trypsin, and the hydrolysing activity was retained well even after trypsin was repeatedly used up to five times. Interestingly, the phosvitin phosphopeptides prepared with immobilised trypsin demonstrated a lower N/P ratio and a higher calcium-binding efficiency than those prepared with free trypsin. Furthermore, phosphopeptides significantly increased the rate of calcium absorption and serum calcium content in vivo. Based on these results, we conclude that trypsin immobilised onto chitosan has a greater phosvitin hydrolysing activity than free trypsin, and the prepared phosphopeptides can be used as a new calcium supplement to significantly increase calcium absorption in growing rats.

Potential of Food Hydrolyzed Proteins and Peptides to Chelate Iron or Calcium and Enhance their Absorption

“…CPPs are commercially available for use in milk-based formula as dietary carriers of calcium and other divalent minerals due to the presence of a unique cluster of three phosphorylated serine residues and two glutamate residues [ 60 ]. Egg yolk also contains phosphorylated protein, like phosvitin, which was used to produce phosphopeptides with a binding capacity towards calcium and improve bioavailability [ 61 ].…”

Phosvitin Derived Phospho-Peptides Show Better Osteogenic Potential than Intact Phosvitin in MC3T3-E1 Osteoblastic Cells

“…Since enzymatic hydrolysis to generate phospho-peptides would likely avoid the perceived pitfalls of native phosvitin, PPPs are a possible approach to enhancing osteogenic and/or osteoprotective functions. Previous studies have also suggested that PPPs show enhanced bioactivities (anti-inflammation, antioxidant and calcium absorption promoting) compared to native phosvitin [ 43 , 44 ], further supporting the role(s) of phosphorylation in bioactive properties. In contrast, bioactive peptides lacking of phosphorylation, such as LRW derived from pea protein [ 45 ], VLPVPQK, EDVPSER, NAVPITPTL, HPHPHLSF derived from buffalo casein [ 46 ], YVEEL and YLLF derived from whey proteins [ 47 ], and IRW from egg proteins [ 48 ], were also reported to promote the proliferation of MC3T3-E1 cells, but they may not be as resistant to digestion as phospho-peptides, which therefore limits their potential applications [ 42 ].…”

The Formation, Structural Characteristics, Absorption Pathways and Bioavailability of Calcium–Peptide Chelates

“…Serum calcium levels (SCL) can serve as a valid indicator of whether an active substance has a pro-calcium absorption effect. For example, SCL in mice were found to be significantly increased after they were administered calcium phosphopeptide chelates [ 97 ]. Similar results were found in a study about the bioactivities of calcium herring egg phosphopeptides chelates.…”