| China's marine shellfish resources are abundant,but the traditional utilization mode has a low output value,and improper handling of its waste materials is likely to cause problems such as resource waste and environmental pollution.On the other hand,affected by traditional eating habits,calcium deficiency in urban and rural residents in China is more common,and the market demand for calcium supplement products is huge.Especially,the complex amino acid chelated calcium of the latest calcium supplement products is very attractive.In this paper,the shellfish processing waste,the scallop skirt,is used as the main raw material to explore the optimal process conditions for the preparation of complex amino acid chelated calcium by enzymatic hydrolysis of scallop skirt protein,and further the preparation of complex amino acid chelated calcium therapy.The effect of osteoporosis is studied by mechanism.The effects of neutral protease,flavor protease,papain and trypsin on the scallop skirt were optimized by single factor and orthogonal test.The free amino acid conversion rate was used as the evaluation index to determine the optimal hydrolysis temperature of each enzyme,time,pH and amount of enzyme added.After optimization,the final free amino acid conversion was 75.25%.The obtained amino acid hydrolysate was combined with calcium chloride,and optimized by single factor and orthogonal test.The chelation rate was evaluated.The optimal chelation reaction temperature was 40 °C,the time was 40 min,and the medium pH was 9.The chelation rate was 92% under the optimum process conditions.Fourier infrared spectroscopy scanning and mass analysis were performed on the obtained complex amino acid chelated calcium to ensure successful calcium ion coupling and product safety.Functional evaluation of the obtained complex amino acid chelated calcium was carried out by animal experiments.In the first-stage animal experiment,SD male rats of four weeks old were used to perform strong bone test.Serum alkaline phosphatase,blood phosphorus,blood calcium,visceral index,calcium metabolism,bone mineral density and other indicators were measured to determine the absorption and utilization efficiency,chronic toxicity and bone-strengthening effect of complex amino acid chelated calcium.In the second phase animal experiment,three-month-old SD castrated female rats were used to simulate postmenopausal osteoporosis for osteoporosis treatment,and serum alkaline phosphatase,serum phosphorus,blood calcium,visceral index,bone mineral density were measured.Estradiol and 25-hydroxyvitamin D and Rank/Rankl/OPG pathway proteins were also detected using the ELISA kit,and WNT/?-Catenin pathway protein was detected by Western Blot.In addition,the analysis was further confirmed using Micro-CT and immunohistochemistry.The results showed that the complex amino acid chelated calcium had better absorption effect,which could significantly increase thebone mineral density of ovariectomized osteoporosis rats,increase the expression of OPG,reduce the expression levels of Rank and Rank1.These effects could inhibit the activity of osteoclasts to achieve the purpose of treating osteoporosis.Meanwhile the complex amino acid chelated calcium is non-toxic and can be safely used.In this study,the preparation process of compound amino acid chelated calcium by scallop skirt was studied,and its efficacy was evaluated in detail.Finally,the conclusion was drawn that the process is simple,green,and environmentally friendly,and the composite amino acid chelated calcium is prepared.The absorption effect is good,and it can treat postmenopausal osteoporosis.It provides basic information for the comprehensive utilization of marine shellfish resources for high value,and has a new idea for the treatment of osteoporosis. |
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