| 摘要: |
| 采用补料-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,在敏隔10 min连续补加36 ml,补料氮溶液(22 s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,类人胶原蛋白表达可达到14 g/L,在4种操作方式中最高。 |
| 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 |
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| 基金项目:国家自然科学基金项目(20476085);国家科技攻关重大专项(2003DA901A32) |
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| Studies on effects of oscillations on cell growth and human-like collagen production of recombinant Escherichia coli |
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| Abstract: |
| By adopting fed-batch fermentation,the effects of nitrogen oscillations on cell growth and target protein formation of recombinant Escherichia coli 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 cultivation 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 C36 mL 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 collagen yield was 14 g/L,the highest in 4 operations. |
| Key words: recombinant Escherichia coli high cell-dcnsity fermentation human like collagen oscillations plasmid stability cell viability |