Preparation of Acaudina leucoproata dual-enzymatic hydrolysates and influence on collagen synthesis of human dermal fibroblasts

To eluicdatie the zymolytic technologcial conditions of Acaudina leucoproata somatic mesoblast, and the effect of hydrolysate on the human dermal fibroblast in vitro, the single factor experiment and orthogonal experiment were adopted to determine the the optimum conditions of the bienzymatic hydrolysis of Acaudina leucoproata somatic mesoblast, and then explore the cell proliferation and collagen production of human dermal fibroblast in vitro via co- culture with the samples though ultrafiltration and freeze drying into powder after enzymolysis.It showed that the optimum conditions were as follows: the ratio of animal proteinase to neutral protease in every sample was 3∶2 by weight, the ratio of distilled water to Acaudina leucoproata was 0.25∶1 ( v /w) , the total enzyme concentration was 4%, enzymolysis temperature was 50℃, enzymolysis time was 4h. After co-culture with CCD-966 SK cells, the result of the present work implied that the samples with different ranges of molecular weight had no cytotoxcity on CCD- 966 SK cells, and the molecular weight range of enzymatic hydrolysates on promoting collagen synthesis and secretion most obviously were below 5ku. The results showed that hydrolysis of Acaudina leucoproata somatic mesoblast by two enzymes could obtain a higher degree of total amino concentration of amino acid.On the other hand, the hydrolysate could accelerate the cell proliferation and collagen production, which established a basis for operation of Acaudina leucoproata.