周期操作对重组大肠杆菌高密度发酵表达类人胶原蛋白的影响  

Studies on effects of oscillations on cell growth and human-like collagen production of recombinant Escherichia coli

作　　者：马晓轩[1] 范代娣[1] 刘树文[2] 骆艳娥[1] 

机构地区：[1]西北大学化工学院,陕西西安710069 [2]西北农林科技大学葡萄酒学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2006年第3期6-10,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(20476085);国家科技攻关重大专项(2003DA901A32)

摘　　要：采用补料一分批式发酵,研究了4种不同间歇周期补氮操作对细胞生长和类人胶原蛋白表达的影响,以及发酵过程中细胞质粒稳定性、细胞菌落形成能力和乙酸生成量的变化。结果表明,在每隔10min连续补加 36mL补料氮溶液(22s完成补加操作)的周期操作下,细胞质粒稳定性明显提高,细胞菌落形成能力下降较小,乙酸生成量低,仅0．7 g／L,最终细胞密度为84 g／L,类人胶原蛋白表达量可达到14 g／L,在4种操作方式中最高。By adopting fed-batch fermentation, the effects of nitrogen oscillations on cell growth and target protein formation of recombinant Escherichia coil producing human like collagen were studied. Four different feeding protocols of nitrogen solution （25 s cycle,5 min cycle,10 min cycle and 20 min cycle） were applied. The total nitrogen fed to the cuhivation was equal in each mode. The cell plasmid stability,the cell viability and acetic acid production during the cultivation were investigated. When the feeding nitrogen solution was added in a way of 10 min cycle （36 mI. feeding nitrogen solution was added to fermentor continuously during 22 s and after 10 min,the second cycle began）,the cell plasmid stability increased. Meanwhile ,the production of acetic acid was small. The final cell density reached 84 g/L. and the human-like col lagen yield was 14 g/L,the highest in 4 operations.

关 键 词：重组大肠杆菌 高密度发酵 类人胶原蛋白 周期操作 质粒稳定性 细胞活性 

分 类 号：Q939[生物学—微生物学]