Preparation of Acaudina leucoproata dual-enzymatic hydrolysates and influence on collagen synthesis of human dermal fibroblasts
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摘要: 探讨双酶水解海地瓜体壁的工艺条件、水解物对人皮肤纤维母细胞的增殖和胶原合成的作用。通过单因素实验以及正交实验确定白肛海地瓜体壁双酶水解的最适宜工艺条件,超滤和冷冻干燥获得的酶解物,与人皮肤纤维母细胞（CCD-966SK）共培养来探讨其对细胞的增殖以及胶原合成的作用。结果显示:在动物蛋白酶与中性蛋白酶的质量比为3∶2、液料比（v/w）为0.25∶1、加酶量4%、酶解温度50℃、酶解时间4h的条件下,酶解效果最好;白肛海地瓜体壁酶解物对细胞无毒性,5ku以内的水解产物对促进CCD-966SK细胞胶原蛋白合成分泌的效果最为明显。结果表明,用双酶水解工艺水解白肛海地瓜体壁获得的游离总氨基酸含量高,对人皮肤纤维母细胞的增殖生长和胶原蛋白的产生具有促进作用。
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关键词:
- 白肛海地瓜 /
- 酶水解物 /
- 人皮肤纤维母细胞（CCD-966SK） /
- 胶原蛋白
Abstract: 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. -
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