Separation And Purification Of Oyster Immune Active Ingredients And Product Development

The immune function of the body is closely related to health,the immune regulation of the balance of the immune response to the body caused by the immune response,a large number of experimental and clinical studies have confirmed that increase the body's immune ability is the most important reason to overcome the disease and restore health.According to the World Health Organization survey results show that the only 5 percent of the world's truly healthy people are in the world,only20 percent of those who are diagnosed with the disease and 75 percent of them are in sub-health.While more than 70% of urban residents in China there are sub-health problems,and present a rising trend year by year the proportion of the sub-health population,,which led to the most essential phenomenond and the most important reason is the body's own immune dysfunction.There are many ways to improve the immunity of the body,but in today's environment of high pressure and fast-paced life does not generality.Bioavailability,safe,non-immune source,and can be directly absorbed by the body of food-borne immunity Active peptide has become a hot topic of attention and research.Oyster is a kind of important economical and edible double shell shellfish,which has been breeding all over the world,mainly in China,Korea,Japan and other countries and regions.The total production of oyster in China is the largest.In 2015,China's oyster yield reached 4.573 million t,which was abundant in resources.High protein content and a complete range of amino acids are good sources for the preparation of immune active peptides.In this paper to research the oysters as raw materialoysters to study the preparation,isolation,purification and activity evaluation of oyster immunocompetent peptides,and the development direction of oyster immune health products was studied.At the same time,the theoretical basis of oyster utilization was provided.,the main research contents and results were as follows:(1)The enzymatic hydrolysis of enzyme and its ultrafiltration components were analyzed by enzymatic hydrolysis.The results showed that the molecular weight distribution of oyster enzymolysis and the ultrafiltration components were mainly concentrated in the <2.5 kDa range,with a content of 53.0%~ 85.0%,and the amino acid composition was complete.Oyster enzymolysis products can promote the spleenlymphocytes and immune function of peritoneal macrophage,the ultrafiltration components(> 10,10 ~ 5,5-2.5 and < 2.5 kDa)on the concentration of 0.25 ~ 4.0mg/mL,the in vitro immune activity intensity has a certain concentration of correlation,and < 2.5 kDa component and 5 ~ 2.5 kDa component of immune function is better than that of other components.The components in the ultrafiltration components of the oyster enzymolysis of the products <2.5 kDa components and5~2.5 kDa components can be used as t the object of further study.(2)The index of immune activity was evaluated by cell experiment and animal experiment.In the range of 0.25 ~ 4.0 mg/mL,the oyster ultrafiltration producton(Mw<5 kDa)could promote the proliferation of splenic lymphocytes and phagocytosis of peritoneal macrophages neutral red and generate NO,and its in vitro immune activity strength has a certain concentration correlation.Animal experiments showed that:the experimental results of lymphocyte transformation and delayed allergic reaction were all positive in the cellular immune function;humoral immune function,serum hemolysin content in the test sample dose group and negative control group no significant difference to the results were negative;non-specific immune function,the influence of the weight of immune organs and the effect of mononuclear macrophage function were all positive.Oyster ultrafiltration producton has a significant effect on the regulation of the immune system.(3)The immunological activity peptide was prepared by separation and purification of oyster ultrafiltration producton(Mw<5 kDa).In vitro immune activity(spleen lymphocyte proliferation activity,phagocytic function of phagocytic cells and NO production of macrophages)were evaluation indicators,using Q Sepharose Fast Flow of strong anion exchange chromatography,Sephadex G-15 gel column chromatography,Superdex Peptide 10/300 column and Symmetyy C18 on oyster ultrafiltration producton were used to purification and selection,finally got a strong immune activity peptide(Mw=430 Da).(4)The product of oyster immune function was developed,and the formula of oyster granule and oral liquid formula was optimized by D-optimal mixing design.The best formula:Oyster granule for the oyster ultrafiltration producton 57.50%,pineapple powder 12.90%,?-cyclodextrin 18.22%,abbas sweet 6.40%,sucrose 5%.Oyster oral liquid(passionfruit)formula for oyster hydrolyzate 97.74%,passionfruit powder0.75%,abbas sweet 0.51%,sucrose 1.0%.Oyster oral liquid(coconut milk)formula was 98.0% of oyster hydrolyzate,0.65% of coconut milk powder,Abbas sweet 0.35%,sucrose 1.0%,and 0.05 g/mL for 0.25 mL citric acid.Oyster peptide granule is loose particles,uniform light yellow,good taste,high protein content,fat is 0%,is a high protein,fat-free quality products.The oyster oral liquid is yellow and milky yellow,and its taste is delicious.It contains high protein and low fat content.Oyster oral liquid and oyster peptide granules are high in trace elements,and their heavy metals and microbial content are in accordance with the national standard.