Purification of Algal Calcium-Chelating Peptide and Its Physical Chemical Properties

The Schizochytrium protein, extracted from byproduct of Schizochytrium sp. after lipid extraction, was isolated and hydrolyzed. A specific calcium-binding peptide was purified from the hydrolysate through Sephadex G-25 gel filtration chromatography and C18 reversed-phase high-performance liquid chromatography (RP-HPLC). The calcium-binding capacity of the specific peptide reached up to 136.68 +/- 4.6 g/mg. The amino acid sequence was confirmed as Ser-Ser-Val (SSV) with a molecular weight of 291.15Da by liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). Fourier transformation infrared spectroscopy showed that the major binding sites included oxygen atom from carbonyl group and nitrogen of amino group or imino group. SSV-Ca chelate was confirmed to be a neutral molecular by Zeta potential analysis, and the calcium ion was surrounded by the functional chelating sites of SSV. In addition, thermogravimetry-differential scanning calorimetry (TG-DSC) and calcium releasing assay revealed that the SSV-Ca chelate exhibited excellent thermal stability and solubility in both acidic and basic conditions, which was in favor for calcium absorption in the gastrointestinal tracts of humans. The findings indicate the by-product of Schizochytrium sp. is a promising source for making peptide-calcium chelate as a dietary functional supplement.