响应面法优化酸溶性鳗鱼鱼鳔胶原蛋白提取工艺

Optimized Extraction of Acid-soluble Collagen From Eel Swim Bladder by Response Surface Methodology

【目的】研究酸法提取鳗鱼(Muraenesox cinereus)鱼鳔胶原蛋白的最佳工艺。【方法】以鳗鱼鱼鳔为研究原料,在单因素试验基础上,应用响应面法研究醋酸浓度、料液比以及酸解时间对提取酸溶性鳗鱼鱼鳔胶原蛋白的影响;并对所提取的胶原蛋白采用紫外光谱(UV)、傅立叶红外光谱(FTIR)检测、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和扫描电镜(SEM)等方法进行理化性质分析。【结果】最佳提取工艺为醋酸浓度0.42 mol/L、酸解时间21 h、料液比1∶58 g/mL,在该条件下鳗鱼鱼鳔胶原蛋白的提取率为65.32%。在190~400 nm的波长范围检测中,波长225 nm处出现强烈的特征吸收峰,波长280 nm处无明显的特征吸收峰;红外光谱检测该胶原蛋白具有三级结构的特征吸收峰;SDS-PAGE检测可见2条α链与1条β链结构;氨基酸分析表明甘氨酸、丙氨酸、谷氨酸、脯氨酸含量较高,分别为19.88%、11.78%、10.48%和9.37%,且不含色氨酸;扫描电镜下的鱼鳔胶原蛋白为均匀致密的纤维网络结构。【结论】酸法提取的鳗鱼鱼鳔胶原蛋白为Ⅰ型胶原蛋白。

【Objective】To study the optimum extraction process of eel swim bladder collagen by acid method.【Methods】Based on single-factor experiments,the effects of acetic acid concentration,material-liquid ratio and acid hydrolysis time on the extraction of acid-soluble collagen from eel swim bladder were studied by response surface methodology(RSM).The extracted collagen was analyzed by Ultraviolet-visible spectroscopy(UV),Fourier infrared spectroscopy(FTIR),Sodium lauryl sulfate Polyacrylamide gel electrophoresis(SDS-PAGE)and scanning electron microscopy(SEM).【Results】The optimum extraction conditions were acetic acid concentration of 0.42 mol/L,acid hydrolysis time of 21 h,material-to-liquid ratio of 1∶58 g/mL.Under these conditions,the extraction rate of eel swim bladder collagen was 65.32%.In the wavelength range of 190-400 nm,a strong characteristic absorption peak appeared at the wavelength of 225 nm,and no obvious characteristic absorption peak appeared at the wavelength of 280 nm The amino acid analysis showed that the contents of glycine,alanine,glutamic acid and proline were 19.88%,11.78%,10.48%and 9.37%,respectively,under the scanning electron microscope,the collagen in the swim bladder is a uniform and dense fiber network structure.【Conclusion】The collagen extracted from the swim bladder of eel by acid method is type I collagen.