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过程工程学报 ›› 2016, Vol. 16 ›› Issue (4): 654-659.DOI: 10.12034/j.issn.1009-606X.216139

• 生化工程专栏 • 上一篇    下一篇

牛关节软骨II型胶原的分离纯化与鉴定

李赛娜1,康跻耀2,邓平晔3,高建萍1,张贵锋2,王明林1   

  1. 1. 山东农业大学食品科学与工程学院
    2. 中国科学院过程工程研究所生化工程国家重点实验室
    3. 北京理化分析测试中心
  • 收稿日期:2016-02-17 修回日期:2016-03-23 出版日期:2016-08-20 发布日期:2016-10-10
  • 通讯作者: 李赛娜 seine2015@163.com

Separation and Characterization of Type II Collagen from Bovine Articular Cartilage

LI Sai-na 1,KANG Ji-yao 1,DENG Ping-ye 2,GAO Jian-ping 1,ZHANG Gui-feng 3,WANG Ming-lin 1   

  1. 1. College of Food Science and Engineering, Shandong Agricultural University
    2. Beijing Center for Physical and Chemical Analysis
    3. State Key laboratory of Biochemical Engineering, Institute of Process Engineering, CAS
  • Received:2016-02-17 Revised:2016-03-23 Online:2016-08-20 Published:2016-10-10
  • Contact: LI Sai-na seine2015@163.com

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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 kDa，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.

关键词: II型胶原, 分离, 生物质谱, 特征多肽

Abstract: The bovine articular cartilage was pretreated using NaCl 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 kDa. The content of Gly in amino acid composition of type II collagen 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 pH 7.0. Properties of collagen purified from bovine cartilage corresponded to bovine type II collagen.

Key words: collagen type II, separation, mass spectrometry, marker peptides

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