摘要
以羊肌腱为实验材料,经脱脂、脱钙、除杂蛋白后,用盐酸胍去除蛋白多糖,经胃蛋白酶酶解、氯化钠盐析、透析后对其进行提取工艺的研究与鉴定。结果表明,单因素试验及正交试验确定最佳工艺条件为酶解时间60 h、酶添加量4%、0.1 mol/L柠檬酸溶液添加量5 mL/g进行羊肌腱提取。紫外光谱分析结果显示:所提胶原最大吸收峰220 nm,证明提取物为胶原蛋白。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果表明:羊肌腱中存在的蛋白主要是Ⅰ型胶原蛋白。经红外光谱分析,进一步证明提取的胶原蛋白为Ⅰ型胶原蛋白且结构完整。
Type Ⅰ collagen from sheep tendon was prepared by the following sequential steps：degreasing and decalcification of the starting raw material,removal of impurity proteins,removal of proteoglycans by extraction with guanidinium hydrochloride,pepsin-catalyzed hydrolysis,salting out with sodium chloride and dialysis.The optimal conditions for enzymatic hydrolysis were determined by single-factor experiments and orthogonal array design experiment as hydrolysis at an enzyme concentration of 4% for 60 h in 0.1 mol/L citric acid with a solvent to substrate ratio of 5：1 mL/g.UV spectral analysis of the resultant extract showed maximum absorption at approximately 220 nm,confirming it to be collagen.The protein composition of sheep tendon was dominated by type Ⅰ collagen,as demonstrated by SDS-PAGE analysis.The extract was further confirmed by infrared spectral analysis to be type Ⅰ collagen with intact structure.
出处
《肉类研究》
2014年第1期8-11,共4页
Meat Research
基金
内蒙古自治区自然科学基金项目(2011MS1208)
关键词
Ⅰ型胶原蛋白
提取工艺
鉴定
type Ⅰ collagen
extraction process
identification
