日本鳗鱼皮胶原蛋白提取工艺优化及理化性质分析

Optimization of Japanese eel skin collagen extraction process and analysis of physicochemical properties

利用响应面法对日本鳗鱼(Anguilla japonica)皮胶原蛋白的提取工艺进行优化,并对其理化性质和致敏性进行研究。采用酸酶浸提法提取鱼皮胶原蛋白,基于单因素试验,以料液比、提取液浓度和提取时间作为考察因素,以胶原蛋白得率为指标,得到最佳提取工艺:料液比1∶30 g/mL,提取液浓度0.6 mol/L,提取时间48 h,胶原蛋白得率为32.89%。将优化后得到的胶原蛋白进行理化性质表征分析,SDS-PAGE结果表明,胶原蛋白含有5个条带,属于Ⅰ型胶原蛋白;必需氨基酸和药用氨基酸占比分别为19.61%和11.04%;傅里叶红外光谱(FTIR)光谱显示其保留了完整的三螺旋结构;在230 nm处有最大的紫外吸收峰;由扫描电镜(SEM)显示胶原蛋白的微观结构呈现出纤维状结构;胶原蛋白的热转变温度和热变性温度分别为38.5和28.3℃;酶联免疫吸附反应(ELISA)和模拟胃肠液实验可知胶原蛋白具有一定的致敏原性,为鳗鱼皮胶原蛋白的进一步研究提供理论基础。

The extraction process of Japanese eel skin collagen was optimized using response surface methodology,and its physicochemical properties and allergenicity were investigated.Based on the singlefactor experiment,the best extraction process was obtained based on the acid-enzymatic extraction method,with the material-liquid ratio,the concentration of extraction solution and the extraction time as the investigating factors and the collagen yield as the index.The results showed that the collagen contained five bands and belonged to type I collagen,and the percentages of essential amino acids and medicinal amino acids were 19.61% and 11.04%,respectively.The thermal transition temperature and thermal denaturation temperature of collagen were 38.5 and 28.3 ℃,respectively.The enzyme-linked immunosorbent reaction and simulated gastrointestinal fluid experiments showed that collagen has certain allergenic properties,providing a theoretical basis for further studies on eel skin collagen.