To produce an easily absorbed calcium supplement and to increase the added value of catfish bone, the optimum enzymatic hydrolysis parameters for preparing catfish bone collagen polypeptide were optimized by single-factor tests with calcium-binding activity as evaluation indexes. The Sephadex G-25 column was performed to enrich the collagen polypeptides with high calcium binding activity. The structure of collagen polypeptide-calcium chelates was investigated by infrared spectroscopy, scanning electron microscopy and energy spectrum analysis. Furthermore, the thermal stability, acid-base stability, and digestion stability in vitro were also evaluated. Results showed that the optimum enzymatic hydrolysis parameters were a material-liquid ratio of 1:13 (g:mL), alkaline protease addition of 4% (mass fraction), hydrolysis pH of 8.0, hydrolysis temperature of 40 ℃, and hydrolysis temperature of 2 h. The achieved collagen polypeptides possessed a calcium-binding capacity of 29.37 mmol/L and the collagen peptides with a molecular mass ranging from 195 to 1 967 Da account for 77.73% of the whole catfish bone collagen peptides. Three fractions with different molecular weights were obtained after column fractionation, among which F3 exhibited the highest calcium binding activity. The amino, carboxyl, and amide bonds of collagen peptides were the main binding sites for Ca2+. Upon chelating with calcium, the smooth plane of collagen polypeptide became rough, loose, and porous, the calcium content was increased. Stability analysis revealed that catfish bone collagen polypeptide-calcium chelates showed certain resistance to heat, weak alkali, and digestion. Over 70% of calcium was retained after gastrointestinal digestion, suggesting high calcium bioavailability. But it was unstable in a strong acid environment. This study could enrich the kinds of calcium supplements and provide technical references for the high-value utilization of fish bone resources.
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