新型重组人肿瘤坏死因子工程菌的发酵工艺研究

Fermentation research of new type recombinant human tumor necrosis factor alpha

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。

PurposeThe aim is to establish fermentation process of new type recombinant human necrosis factor (nrhTNF)α.MethodsThe optimized host cell,culture medium and induction time were deternined according to the growth and expression specificity of nrhTNF on test tube and flask shaker.Then fed batch culture was carried out on 5 L automatic fermentor.ResultsThe expression of nrhTNF in fermentor could keep in a higher level of about 50% of total bacterial proteins when fermented to around A 600nm 30,with a fermentation time of 12 h to 13 h.ConclusionA short cycle,high expression and stabilized fermentation process was established.