鲨鱼软骨胶原蛋白的酶法制备及其稳定性研究

Study on Enzymatic Preparation and Stability of Shark Cartilage Collagen

以鲨鱼软骨为原料,通过单因素实验和响应面试验优化酶法制备胶原蛋白的最佳工艺,并分析对胶原蛋白的结构和稳定性。结果表明,胶原蛋白提取的最佳条件为:胃蛋白酶添加量9150 U/g、提取时间39.85 h、pH 1.67及料液比1∶15(g/mL),此条件下胶原蛋白的提取率为(85.09±0.17)%,所制备的胶原蛋白保留了较为完整的三螺旋结构,符合Ⅱ型胶原蛋白的特征。稳定性研究表明其变性温度为39.9℃,黏度随温度和盐浓度的升高显著下降,而随pH值的不同呈先增后减再增加的趋势,以pH 7时黏度最小(4.26 mPa·s)。圆二色谱分析表明,随盐浓度和温度的增加,胶原蛋白的三螺旋结构发生改变,当温度超过变性温度时(39.9℃),三螺旋结构逐渐解旋且变性。本研究为鲨鱼资源的高值化利用提供了理论依据。

Using shark cartilage as raw material,the optimal enzymatic preparation process of collagen was optimized through single factor and response surface experiments,and the structure and stability of the prepared collagen were analyzed.The results showed that the optimal conditions of extraction collagen were as follows:9 150 U/g of pepsin,39.85hours of extraction time,pH 1.67,and a ratio of 1∶15(g/mL).Under these conditions,the extraction rate of collagen was(85.09±0.17)%,and the prepared collagen retained a relatively complete triple helix structure,which conforms to the characteristics of type Ⅱ collagen.The stability study showed that the denaturation temperature of the prepared collagen was 39.9 ℃,and its viscosity decreased significantly with the increase of temperature and salt concentration,but there was no linear relationship between the viscosity and pH value,and its viscosity was the smallest at pH 7(4.26mPa·s).Circular dichroism analysis further indicated that with the increase of salt concentration and temperature,the triple helix structure of collagen changed,when the temperature exceed the denaturation temperature of 39.9 ℃,the tripartite helix structure gradually unrotated and denatured.This study provides a specific theoretical basis for the high-value utilization of shark resources.