文章摘要
刘雯恩,甘兵,范志强,李玉玲,彭新生,周艳芳.优化I型胶原蛋白的纯化工艺[J].精细化工,2019,36(5):
优化I型胶原蛋白的纯化工艺
Optimization of purification process of collagen type I
投稿时间:2018-10-12  修订日期:2019-02-03
DOI:
中文关键词: I型胶原蛋白  等电点沉淀法  超滤纯化  圆二色谱法  生物工程
英文关键词: Collagen type I  Isoelectric precipitation method  Ultra-filtration purification  Circular Dichroism  biological engineering
基金项目:广东省科技计划项目(2015A020214020);东莞市科技计划重点项目(2017507152431)
作者单位E-mail
刘雯恩 广东医科大学  
甘兵 广东医科大学 第三附属医院  
范志强 广东医科大学  
李玉玲 广东医科大学  
彭新生 广东医科大学  
周艳芳 广东医科大学 fangfang772003@163.com 
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中文摘要:
      本文使用0.5mol/L乙酸和胃蛋白酶提取I型胶原蛋白后,结合等电点沉淀法和超滤纯化技术对I型胶原蛋白进行纯化。通过鞣酸沉淀法和聚丙烯酰胺凝胶电泳(SDS-PAGE)证实等电点沉淀可有效除去胃蛋白酶;经超滤纯化后的胶原蛋白紫外光谱检测在235nm处有最大吸收峰;在SDS-PAGE谱图上出现3条I型胶原蛋白特征谱带;采用凯氏定氮法也证实其纯度符合YY/T 1511-2017医药行业标准;通过傅里叶变换红外光谱、圆二色谱法(Circular Dichroism, CD)测定证实胶原蛋白三螺旋结构被完整保留下来。采用等电点沉淀法结合超滤纯化技术,可保证I型胶原蛋白的高纯度及三螺旋结构的完整性。
英文摘要:
      In this study, isoelectric point precipitation method and ultra-filtration purification technique was used to purify the collagen type I extracted from bovine tendon by using acetic acid and pepsin with low temperature. Tannic acidprecipitation and SDS-PAGE gel electrophoresis demonstrated that isoelectric precipitation could effectively remove pepsin; The purified collagen type I had a maximum absorption peak at 235 nm in Ultraviolet spectroscopy; SDS-PAGE spectra showed three characteristic bands of collagen type I; The Kjeldahl method also confirmed that the collagen purity was in line with the standard(YY/T 1511-2017); Fourier transform infrared spectroscopy andcircular dichroism confirmed that the triple helix structure of collagen was completely preserved. The data means the high purity of collagen type I and the integrity of the triple helical structure can be insured by the isoelectric precipitation method combined with ultra-filtration purification technique.
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