表达重组人生长激素的大肠杆菌高密度发酵的初步试验研究

Primary Studies on High Cell Density Fermentation of Recombinant Escherichia Coli Expressing Human Growth Hormone

根据表达重组人生长激素的大肠杆菌的自身特点 ,对其发酵的工艺和条件作了初步的研究和改进 ,通过以甘油代替葡萄糖作为培养基中的碳源 ,以流加的方式在发酵过程中逐步补加 ;同时 ,通过控制搅拌转速 ,以控制溶氧不低于 2 0 %的水平 ,在发酵的中后期 ,再以流加的方式补充适量的蛋白胨等营养成份 ,使发酵终止时 ,大肠杆菌的密度由 30 g/L提高到 90 g/L,而 r- h GH的细胞表达量并未因此而受到影响 ,发酵周期从 16 h缩短到 10 h。

According to the characterization of recombinant Escherichia coli expressing human growth hormone, We made some primary studies and progress on the methods and conditions of its fermentation The new method was developed which, by means of glycerol being the carbon sources instead of glucose and its feeding by means of a specific fed batch mode; in the meantime, by controlling the stirrer speed and keeping the dissloved oxygen (pO2) level at 20% saturation; feeding proper nitrogen sources (such as tryptone) by means of fed batch mode in the later phase of fermentation; allows growth of the recombinant Escherichia coli cell density from 30 g/L up to 90 g/L Meanwhile, the product of r hGH keeps high level and the fermentation time is reduced from 16 hours to 10 hours