Improvement of hEGF production with enhanced cell division ability using dissolved oxygen responses to pulse addition of tryptone
Improvement of hEGF production with enhanced cell division ability using dissolved oxygen responses to pulse addition of tryptone
复制标题
利用溶解氧对胰蛋白胨脉冲添加的反应来提高细胞分裂能力，从而改善 hEGF 的产生
DOI：
10.1007/s12257-008-0052-1
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发表时间：
2009-03
影响因子：
3.2
通讯作者：
Zheng, Zhi-Yong
中科院分区：
文献类型：
--
作者：
Zhan, Xiaobei;Yao, Shan-Jing;Lin, Chi Chung;Zheng, Zhi-Yong
在重组大肠杆菌的脉冲流加分批培养中，胰蛋白内酯对细胞生理和工艺性能的影响是多方面和复杂的。通过在进料速度中应用溶氧信号对脉冲响应的反馈控制，避免了乙酸乙酯的产生，成功地实现了重组人表皮生长因子(HEGF)的生产优化。在脉冲补料分批培养中，添加适量的胰蛋白糖和葡萄糖。在大肠杆菌中，细胞分裂能力和细菌内质粒的稳定性都得到了提高。因此，细胞分离成可存活但不可培养的生理状态的情况得到了缓解。在hEGF表达前后加入胰蛋白调也能增强细胞的呼吸作用，从而有利于重组hEGF的产生。过量添加胰蛋白胨导致溶氧信号振荡灵敏度低，脉冲补料分批培养的可操作性差，导致醋酸盐积累，削弱了溶氧信号的响应。因此，重组蛋白的产量大大减少。结合工艺性能和复合介质脉冲添加对细菌代谢的积极作用，成功地确定了hEGF的最佳生产条件。在此最佳生产条件下，菌体密度可达91g/L干重。此外，0.24g/L人表皮生长因子的高水平使产品收率比对照提高了32.6%。
Tryptone has multiple and complex effects on cell physiology and process performance in pulse fed-batch cultivation of recombinantEscherichia coli. By applying feedback control of dissolved oxygen signal responding to pulse in the feed rate, the production of acetate was avoided and the optimization of production of recombinant human epidermal growth factor (hEGF) was successfully achieved. With the addition of an optimum amount of tryptone along with glucose in the pulse fedbatch cultivation ofE. coli, the ability of the cell to divide and the stability of the plasmid within the bacteria were improved. Consequently, segregation of the cells into a viable but non-culturable physiological state was alleviated. Addition of tryptone also enhanced cell respiration before and after hEGF expression and thus further benefited the production of recombinant hEGF. Excessive addition of tryptone resulted in low sensitivity of the oscillation of dissolved oxygen signal and poor operability of pulse fed-batch cultivation as this led to an accumulation of acetate, which weakened the dissolved oxygen signal responses. Consequently, the production of recombinant protein was considerably reduced. By combining the process performance and the positive effect of complex media pulse addition on bacterial metabolism, the optimal production conditions of hEGF were successfully determined. A high cell density of 91 g/L dry cell weight was obtained under these optimal production conditions. Furthermore, a high level of 0.24 g/L hEGF was attained leading to a 32.6% increase in product yield as compared to the controls.
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DOI：
--
发表时间：
1949
期刊：
--
影响因子：
--
作者：
J. Sumner;G. Somers
通讯作者：
J. Sumner;G. Somers
影响因子：
3.2
作者：
Birgit M. Prub;J. Nelms;'. C. Park;J. Alan;Wolfe
通讯作者：
Birgit M. Prub;J. Nelms;'. C. Park;J. Alan;Wolfe
DOI：
10.1016/s0140-6736(81)92814-2
发表时间：
1981-12
期刊：
The Lancet
影响因子：
--
作者：
A. Emery
通讯作者：
A. Emery
影响因子：
3.8
作者：
H. Lin;F. Hoffmann;A. Rozkov;S. Enfors;U. Rinas;P. Neubauer
通讯作者：
H. Lin;F. Hoffmann;A. Rozkov;S. Enfors;U. Rinas;P. Neubauer
影响因子：
3.8
作者：
S. Velut;A. Castan;K. A. Short;J. Axelsson;P. Hagander;Barry A. Zditosky;Christopher W. Rysenga;L. de Maré;Jan-Erik Haglund
通讯作者：
S. Velut;A. Castan;K. A. Short;J. Axelsson;P. Hagander;Barry A. Zditosky;Christopher W. Rysenga;L. de Maré;Jan-Erik Haglund