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为实现重组人胸腺肽α1工程菌E .coliTop10 /pThioHis Tα的高密度高表达发酵 ,首先进行了培养条件的摸索 ,确定了该工程菌的最佳培养条件、诱导剂IPTG的浓度、诱导起始和结束时间 ,然后用 5L自控发酵罐进行分批补料培养 ,发酵中采用分阶段限制性流加氮、碳源 ,保持溶解氧在 35 %左右 ,结果经 3mmol/LIPTG诱导 5h ,3批重复发酵 ,最终菌体密度均达到 6 0A60 0 以上 (相当于干菌 2 5 .6 g/L) ,保持并超过了该重组蛋白在试管和摇瓶中的表达量 ,融合蛋白ThioHis Tα的表达占菌体总蛋白的 42 .4%左右 ,含量达到 5 .7g/L ,相当于胸腺肽α12 .4g/L ,为胸腺肽α1的下游纯化和工业化生产奠定了基础
In order to achieve high-density, high-expression fermentations of the recombinant human thymosin α1 engineering strain E.coliTop10 / pThioHis Tα, the culture conditions were first explored, and the optimal culture conditions of the engineering bacteria, the concentration of the inducer IPTG, the initiation of induction and End time, and then use 5L fed-batch fermentor fed culture, fermentation step by step with limited flow of nitrogen and carbon sources, to maintain dissolved oxygen at about 35%, the results induced by 3mmol / LIPTG 5h, three batches of repeated fermentation , The final cell density reached more than 6 0A60 0 (equivalent to 25.6 g / L dry bacteria), to maintain and exceed the expression of the recombinant protein in vitro and shake flask, the expression of the fusion protein ThioHis Tα accounted for bacteria The total protein of 42.4%, the content of 5 .7g / L, equivalent to thymosin α12 .4g / L, for the thymosin α1 downstream purification and industrialization laid the foundation