该研究利用醋酸和胃蛋白酶结合从乌鳢鱼皮中提取胶原蛋白。采用单因素试验和正交试验考察了醋酸浓度、料液比、酶用量、提取时间等条件对胶原蛋白提取率的影响。结果表明,当醋酸的浓度为0.5 mol/L, 胃蛋白酶的添加量为鱼皮质量的1%,提取时间48 h,料液比1∶70,胶原蛋白提取率达到51.2%。通过氨基酸组成分析、圆二色谱、聚丙烯酰胺凝胶电泳等方法对胶原蛋白的结构进行测试,发现该法提取的胶原蛋白具有典型Ⅰ型胶原蛋白特征,其三螺旋结构保持完整,分子质量约为340 kDa。成品中蛋白质含量> 95%, 羟脯氨酸的含量> 5%,灰分<1%(均为质量分数),可用于医用材料、化妆品、功能食品等领域。
In this research, collagen was extracted from the skin of snakehead fish by the combination of acetic acid and pepsin. The effects of acetic acid concentration, solid-liquid ratio, enzyme dosage, and extraction time on the extraction rate of collagen were investigated by single factor experiment and orthogonal test. Results showed that when the concentration of acetic acid was 0.5 mol/L, the addition of pepsin was 1% of the weight of fish skin, the extraction time was 48 h, and the solid-liquid ratio was 1∶70, the extraction rate of collagen reached 51.2%. The structure of collagen was tested by amino acid composition analysis, circular dichroism, and polyacrylamide gel electrophoresis. It was found that the collagen extracted by this method had typical type I collagen characteristics, its 3-helix structure remained intact, and its molecular weight was about 340 kDa. The protein content of the final goods was more than 95%, the hydroxyproline content was more than 5%, and the ash content was less than 1%, which could be used in medical materials, cosmetics, functional foods and other fields.
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