皮革科学与工程 ›› 2022, Vol. 32 ›› Issue (1): 9-14.doi: 10.19677/j.issn.1004-7964.2022.01.002

• 试验研究 • 上一篇    下一篇

非变性牦牛骨胶原蛋白的提取与表征

么林妍1, 凌碧扬1, 刘晓乐2, 刘茵3, 肖建喜1,*   

  1. 1.兰州大学化学化工学院,甘肃 兰州 730000;
    2.甘肃省化工研究院有限责任公司,甘肃 兰州 730000;
    3.甘肃创翼检测科技有限公司,甘肃 兰州 730000
  • 收稿日期:2021-05-31 出版日期:2022-02-28 上线日期:2022-01-13
  • 通讯作者: *xiaojx@lzu.edu.cn
  • 作者简介:么林妍(1993-),女,博士,主要从事胶原蛋白的研究工作,yaoly17@lzu.edu.cn。
  • 基金资助:
    甘肃省重点研发计划(18YF1NA004)

Extraction and Characterization of Non-denatured Yak Ossein

YAO Linyan1, LING Biyang1, LIU Xiaole2, LIU Yin3, XIAO Jianxi1,*   

  1. 1. College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000, China;
    2. Gansu Research Institute of Chemical Industry Co. , Ltd. , Lanzhou 730000, China;
    3. Gansu Chuangyi Testing Technology Co. , Ltd. , Lanzhou 730000, China
  • Received:2021-05-31 Online:2022-02-28 Published:2022-01-13

摘要: 三股螺旋结构是胶原蛋白的关键结构特征,胶原蛋白变性会破坏三股螺旋结构进而严重影响其生物功能。牦牛是高寒地区特有的半野生牛种,主要生活在我国的青藏高原。我们开发了一种温和的生物工艺,从牦牛骨中提取制备了高品质的牦牛胶原蛋白(Yak ossein,YO)。聚丙烯酰胺凝胶电泳(SDS-PAGE)和圆二色谱(CD)结果表明,我们提取得到的为I型牦牛胶原蛋白,未发生降解,并且整体保持完整的三股螺旋结构。酶切实验进一步表明,提取的YO未发生变性,它可以有效抵抗胃蛋白酶的降解。同时,我们建立变性胶原蛋白的靶向结合实验,利用高特异性的荧光多肽探针来检测牦牛胶原蛋白的变性程度。该靶向探针对提取的YO完全没有结合能力,表明该YO为非变性胶原蛋白。细胞实验表明,YO对成纤维细胞有良好的粘附能力,并且可以显著促进其细胞增殖。

关键词: 牦牛骨胶原蛋白, 三股螺旋结构, 变性胶原蛋白, 靶向探针

Abstract: The triple helix structure is the key feature of collagen, and the denaturation of collagen will destroy the triple helix structure and seriously affect its biological functions. Yak is a semi-wild cattle species unique to alpine regions, and it mainly lives in the Qinghai-Tibet Plateau. We developed a mild biological process to extract high-quality yak collagen from yak bones. Polyacrylamide gel electrophoresis (SDS-PAGE) and circular dichroism (CD) results showed that the extracted type I yak collagen was not degraded, and the triple helix structure was maintained. The enzyme digestion experiment further showed that the extracted Yak ossein was not denatured, and it can effectively resist the degradation by pepsin. Meanwhile, we established a targeted binding experiment of denatured collagen, using highly specific fluorescent peptide probes to detect the denaturation level of yak collagen. The targeting probe had no binding ability to the extracted Yak ossein, indicating that the Yak ossein is non-denatured collagen. Cell experiments showed that Yak ossein had good adhesion to fibroblasts and can significantly promote cell proliferation.

Key words: Yak ossein, triple helix, denatured collagen, targeting probe

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