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在 10L发酵罐中对戊型肝炎病毒衣壳蛋白在重组大肠杆菌中表达发酵工艺进行了研究 ,用分批培养方法探讨了不同培养基、培养基中磷酸盐浓度和Mg2 + 浓度等因素对菌体生长与重组蛋白表达的影响 ;用分批补料培养研究了不同的补料工艺对菌体生长与重组蛋白表达的影响 ,同时对重组菌诱导时期、诱导持续时间以及不同诱导温度表达包含体在尿素溶液中的溶解性进行了研究。结果表明 ,在优化后的培养基中 ,磷酸盐浓度、Mg2 + 浓度分别为 80mmol L与 2 0mmol L时菌体生长与表达效果较好 ;分批补料培养中 ,37℃培养 9h菌体达到对数期中期 (约 4 5OD6 0 0 )为适宜诱导时期 ,加入终浓度为 1 0mmol LIPTG后诱导 5h ,OD6 0 0 达到 80以上 ,重组蛋白表达量达到2 9 74 % ,为最适收获菌体时间 ;37℃表达的包含体 80 %以上溶解在 4mol L的尿素溶液中 ,最终浓度达到 14mg mL ;10L发酵罐中确定的发酵工艺参数在 30L发酵罐中进行了放大培养 ,10L发酵罐中确定的发酵工艺参数在 30L发酵罐上具有可放大性与重复性 ,可以应用于工业生产。
In 10L fermentor of hepatitis E virus capsid protein expression in recombinant E. coli fermentation process were studied using batch culture medium to explore the different media, phosphate concentration in the medium and the concentration of Mg2 + and other factors on the bacteria Body growth and recombinant protein expression. The effect of different feeding processes on cell growth and recombinant protein expression was studied by fed-batch culture. The effects of different feeding processes on the growth of recombinant bacteria, the duration of induction, Solubility in urea solution was studied. The results showed that in the optimized culture medium, the growth and expression of the cells were better when the concentration of phosphate and Mg2 + were 80 mmol L and 20 mmol L, respectively. In the fed-batch culture, The mid-logarithmic phase (about 405D60) was suitable for the induction period. After the addition of 10mmol / L IPTG for 5h, the OD600 reached 80 and the recombinant protein reached 29.9% 80% of the inclusions expressed at 37 ℃ were dissolved in 4mol L urea solution and the final concentration reached 14mg mL. The fermentation parameters determined in the 10L fermenter were amplified and cultured in a 30L fermenter, The fermentation process parameters in the 30L fermenter with scalability and repeatability, can be used in industrial production.