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目的研究并建立新型重组人肿瘤坏死因子 (nrhTNF)的发酵工艺。方法通过探讨nrhTNF工程菌在试管和锥形摇瓶中的生长和表达规律 ,筛选并确定其最适宿主菌、最佳培养基及最佳诱导表达条件 ,然后在 5L自控发酵罐中进行分批补料培养。结果nrhTNF工程菌在 5L发酵罐中培养至终密度A60 0nm30左右时 ,目的蛋白的表达最高 ,保持在菌体总蛋白的 5 0 %附近 ,发酵时间为 12～ 13h。结论确定了周期短、产率高且稳定可靠的发酵工艺 ,为nrhTNF的工业化生产奠定了基础。
Objective To study and establish a novel recombinant human tumor necrosis factor (nrhTNF) fermentation process. Methods The growth and expression patterns of nrhTNF engineered bacteria in test tubes and conical shake flasks were investigated. The optimal host strains, the optimal culture medium and the best induction conditions were screened and determined. Then the batches were batched in a 5L autocontrol fermenter. Feeding culture. Results The nrhTNF engineered bacteria were cultured in a 5L fermentor to a final density of A60 0nm30, and the expression of the target protein was highest, which was maintained at around 50% of the total bacterial protein. The fermentation time was 12 to 13 hours. Conclusion The fermentation process with short cycle, high yield and stable and reliable was established, which laid the foundation for the industrial production of nrhTNF.