金鲳鱼鱼皮酶溶性胶原蛋白的提取工艺优化及基本特性
Optimization of extraction process and basic properties for pepsin-soluble collagen from Golden pompano skin
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摘要: 为提高金鲳鱼加工副产物——鱼皮的利用率,本研究在前期单因素实验的基础上,以加酶量、液料比和提取时间三个单因素建立模型,采用响应面法Box-Behnken设计实验,优化了胃蛋白酶法提取金鲳鱼鱼皮酶溶性胶原蛋白(PSC)的提取工艺,并与酸溶性胶原蛋白(ASC)进行聚丙烯酰胺凝胶电泳、傅里叶变换红外光谱和差示扫描量热仪等基本理化特性的对比研究。结果表明,金鲳鱼鱼皮胶原蛋白的最佳提取工艺为:加酶量1.5%、液料比23∶1 m L/g、酶解时间27.5 h。此条件下通过实验验证得出,金鲳鱼鱼皮胶原蛋白实际提取率为64.68%,与响应面模型理论预测值65.17%基本一致;此外,本实验提取的PSC有较高热变性温度(30.44℃),与金鲳鱼皮ASC有相似的电泳性质和红外峰型,初步鉴定为I型胶原蛋白。该实验为金鲳鱼鱼皮酶溶性胶原的提取提供了理论参考。Abstract: To improve the utilization rate of fish skin in processing by-product of Golden pompano, this paper used the response surface Box-Behnken method to optimize the extraction process of pepsin-soluble collagen ( PSC) from the skin of Golden pompano.Based on the previous single factor experiment, the model was established by three factors ( enzyme concentration, liquid-to-solid ratio and extraction time) .In addition, the basic properties of PSC were characterized and compared with acidsoluble collagen ( ASC) by Sodium Dodecyl Sulfate Poly Acrylamide Gel Electrophoresis ( SDS-PAGE) , Attenuated Total Reflectance Fourier Transform Infrared spectroscopy ( ATR-FTIR) and Differential Scanning Calorimeter ( DSC) . The results showed that the optimum extraction conditions were 1.5% enzyme concentration, 23∶ 1 m L/g liquid-to-solid ratio, and 27.5 h extraction time.Under this condition, the actual yield of collagen of Golden pompano skin was 64.68%, which was close to the predicted value ( 65.17%) .Moreover, the PSC obtained in this experiment had higher denaturation temperature ( 30.44 ℃) and preliminarily identified as type I collagen, which was similar to the electrophoretic and infrared peaks of ASC. The technical reference for the extraction of PSC from Golden pompano skin was given in this research.