牛关节软骨Ⅱ型胶原的分离纯化与鉴定  

Separation and Characterization of Type Ⅱ Collagen from Bovine Articular Cartilage

作　　者：李赛娜 康跻耀[2] 邓平晔 高建萍[2] 张贵锋[2] 王明林[1] 

机构地区：[1]山东农业大学食品科学与工程学院,山东泰安271018 [2]中国科学院过程工程研究所生化工程国家重点实验室,北京100190 [3]北京理化分析测试中心,北京100089

出　　处：《过程工程学报》2016年第4期654-659,共6页The Chinese Journal of Process Engineering

基　　金：广东省自然科学基金资助项目(编号:2014A030312013);国家高技术研究发展计划(863)基金资助项目(编号:2014AA022109)

摘　　要：将牛关节软骨用NaCl和盐酸胍预处理,在酸性条件下用胃蛋白酶降解杂蛋白,经盐析、透析和冷冻干燥后获得Ⅱ型胶原,对其进行鉴定.结果表明,提取的Ⅱ型胶原纯度为92.2%,得率为61.5%,单条链分子量为130 k Da,氨基酸组成中Gly含量超过30%,Pro与Hyp总含量超过20%,具有胶原特有的超分子结构,中性条件下变性温度为43.68℃,符合II型胶原特性.The bovine articular cartilage was pretreated using Na Cl and guanidine hydrochloride to remove impurities and the proteoglycan respectively. Then the miscellaneous proteins were digested by pepsin, and the type Ⅱ collagen was purified by salting out, dialysis and freeze-dried, Type II collagen was characterized. The results showed that a single band（α-chain） with a subunit molecular weight of 130 k Da. The content of Gly in amino acid composition of type Ⅱcollagen was over 30%, and the total content of Pro and Hyp was over 20%. The purified type Ⅱ collagen had unique supramolecular structures of collagen. The denaturation temperature of type II collagen was 43.68℃ in buffer solution with p H 7.0. Properties of collagen purified from bovine cartilage corresponded to bovine type II collagen.

关 键 词：Ⅱ型胶原 分离 生物质谱 特征多肽 

分 类 号：Q819[生物学—生物工程]