Research On High Phosphorylation Of Unionid Peptide-calcium

Our country is a big production country with low utilization rate and a large number of freshwater unionid. As a byproduct of aquaculture, unionid meat has been discarded directly. It is harmful to the growth of fish and environment. The unionid protein with the complete type and reasonable proportion amino acids is a good source of protein. Unionid meat as the raw material, are preparated the active peptide by papain enzyme, phosphorylating and modificating by sodium phosphate polymer. The phosphorylated peptide and calcium ion chelate phosphorylated peptide calcium and then using infrared spectrum to characterization of the structure, finally, the active peptide, the peptide calcium then then poison reason and prevention of osteoporosis. The results showed that:1. By single factor and orthogonal experiment to determine optimum process of unionid peptide: protein concentration was 100mg/ml, the reaction temperature is 60 ?, the pH value of 8, the reaction time is 4h. Under the conditions, the peptide content was 13.35mg/ml.2. Determining the unionid peptide phosphorylation optimum process by single factor and response surface experiment: the reaction temperature is 50.13°C, pH value is 8.41, the reaction time is 1.53 h. Effect of various factors order: pH > reaction temperature > reaction time. At reaction temperature of 50°C, pH value of 8.4, the reaction time of 1.5h the degree of phosphorict is 0.3927g/100 g by validation experiments.3. By studying the peptide calcium ratio, reaction temperature, pH value, reaction time, four factors of phosphorylated peptide then calcium complex reaction, the effect of rock under the best conditions: peptide calcium ratio 1:1, pH 8, the temperature of 50?, 60 min, then are phosphorylated peptide calcium calcium quality score as high as 7.3%.Through infrared spectrum analysis of polypeptide, phosphorylated peptide between calcium ion and then rock reaction.4. The mouse test with different doses of the active peptide and the peptide calcium by HE staining, observed the surface structure morphology and cell morphology of the liver, spleen, kidney. The active peptide and the peptide calcium are did not produce visible pathological and poison rational change in the major organs of mice, having no obvious toxic reason on mice. In rats with osteoporosis prevention experiment; the model group, slow, mental fatigue, hair loss and less feed intake, the weight is generally lower than blank group. After lavaging calcium, the basic health of rat are eased, weight stable rise. Analyzing changes from blood calcium, phosphorusand serum alkaline phosphatase, the groups of calcium preparations have the effect of calcium supplementation, has good effect on prevention of osteoporosis, and the effect of the order then peptide calcium > calcium gluconate > calcium carbonate.