龟皮胶原蛋白的提取及结构表征(Ⅰ)

采用酶法从乌龟皮中提取胶原蛋白,用聚丙烯酰胺凝胶电泳(SDS-PAGE)方法检测龟皮胶原蛋白多肽产品分子量的分布,并将龟皮胶原蛋白和Ⅰ型胶原蛋白样品进行液相色谱分析(HPLC),比较其氨基酸成分和含量.SDS-PAGE电泳结果分析表明龟皮胶原为Ⅰ型胶原蛋白,分子量约为288ku.龟皮胶原蛋白中,甘氨酸的含量最高,占总量的24.69%;其次是谷氨酸、精氨酸和丙氨酸,分别为9.95%、8.08%和9.31%;天冬氨酸、赖氨酸、丝氨酸、苏氨酸、亮氨酸、苯丙氨酸、异亮氨酸、缬氨酸、酪氨酸等含量在5.58%～1.28%之间;组氨酸和蛋氨酸的含量最低,仅占0.79%和0.09%.由于龟皮胶原蛋白的含量高于Ⅰ型胶原蛋白,且与Ⅰ型胶原蛋白的氨基酸含量差异显著,因此,龟皮胶原蛋白可以作为一种重要的皮胶原进行开发和利用.

Extraction and characterization of collagen from turtle-skin

Using emzymic method to distill the turtle-skin collagens from turtles＇ skin;using Polyacrylamide Gel Electrophoresis（SDS-PAGE） to detect the distributing of the molecular mass;using High Performance Liquid Chromatography（HPLC） to analyse the samples of the turtle-skin collagens and typeⅠcollagens and compare their amino acid componets and contents.The collagen of turtle-skin was extracted and studied after pepsin-digestion.The result of the SDS-PAGE showed that the extraction product was type Ⅰ collagen,and its molecule mass was about 288 ku.The compositions of turtle-skin collagen is similar to the components of type Ⅰ collagens.In these 15 kinds of analyzed amino acids,the Gly was the highest content,occupying 24.69% of the amount.The Glu,Arg and Ala was about 9.95%,8.08% and 9.31% respectively.The content of other amino acids ranging from Tyr to Asp was between 5.58%~1.28%.The His and Met were the lowest content,merely occupying 0.79% and 0.09%.Because the content of turtle-skin was higher than that of type Ⅰ collagen and the difference between them was obvious,the turtle-skin collagen could be an important dermomyotome to be developed and used.