摘要
目的研究重组类人胶原蛋白的基因工程下游工艺对阴离子交换树脂(DE52)和阳离子交换树脂(CM52)的吸附效果。方法将基因工程菌E.coliBL21经超声破碎、硫酸铵盐析、超滤浓缩后,再进行批量层析进一步分离纯化。结果得到最佳实验条件采用阴离子交换树脂,pH值为6.0,盐浓度为0.2mol/L,蛋白进样浓度为0.4%(质量分数)。结论在HCB I的纯化过程中,阴离子交换树脂(DE52)较阳离子交换树脂(CM52)的分离效果更优,最终产物的纯度达到95.98%,回收率为91.7%,纯化倍数为5.3。纯化的类人胶原蛋白Ⅰ经SDS-PAGE电泳测定为电泳纯,分子质量为90ku。
Aim To report a large-scale purification method of recombinant human collagen-like bioprotein （HCBⅠ） . Methods Harvested recombinant E. coli cell was lyses by ultrasonic. Supernatant was treated with ammunition sulfate. After centrifugation, the supernatant was cut-off by ultra-filtration, and finally passes through a batch chromatography. Results The process parameters of large-scale batch chromatography were discussed. The DEAE-52 resin is better than CM52 resin as separation media. At the same time, the best operations condition of adsorption and elution of HCB Ⅰ was pH value 3.3, the concentration of NaCl （ ionic strength） 0.2 mol/L and original protein concentration 0.4%. Conclusion The purity of purified human-like collagen has exceeded 95.98%, the recovery rate and purification fold of aim protein were 82.78% and 5.3 respectively. The obtained recombinant human collagen-like protein shows a relative molecular weight of 90 000 in SDS-Page.
出处
《西北大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第6期737-740,共4页
Journal of Northwest University（Natural Science Edition）
基金
国家科技攻关重大专项基金资助项目(2003DA901A32)
国家自然科学基金资助项目(200476085)
关键词
重组类人胶原蛋白Ⅰ
分离纯化
层析
recombinant human-like collagen
separation and purity
chromatography
作者简介
米钰（1968-），男，陕西西安人，西北大学讲师，从事发酵工程与生物材料研究。
