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目的 :探讨发酵条件对大肠杆菌工程菌表达重组人血管抑素 (rhAGN)的影响。方法 :应用摇瓶和发酵罐发酵试验观察不同诱导时机和诱导时间以及 pH、通气量等对rhAGN表达量的影响。结果 :(1)工程菌E .coliDH5α(pBVA2 )在 30℃培养至A60 0nm 为 0 .3～ 0 .5时于 4 2℃诱导4hrhAGN表达量最高 ,约为 2 12mg L ;(2 ) 15L发酵罐发酵参数为 pH7.2 ,搅拌转速 50 0r min ,通气量 4～ 8L min ,溶氧 50 %时 ,rhAGN表达量提高到 2 76mg L。结论 :确定了最适诱导时机和诱导时间 ,通过调节 pH和改善通气 ,初步形成了一套高表达rhAGN的发酵工艺。
Objective: To investigate the effect of fermentation conditions on recombinant human angiostatin (rhAGN) expression in Escherichia coli. Methods : Shake flask and fermenter fermentation experiments were used to observe the effects of different induction time and induction time, as well as pH and ventilation volume on the expression of rhAGN. RESULTS: (1) When A.coli DH5α(pBVA2) was cultured at 30°C to A60 0nm of 0.3-0.5, 4hrh AGN was induced at 4 2°C, and the expression level was the highest, about 2 12 mg L; (2) 15L fermentation. The fermentation parameters of the tank were pH 7.2, stirring speed 50 0 min, ventilation 4-8 L min, dissolved oxygen 50%, rhAGN expression increased to 2 76 mg L. Conclusion : The optimum induction time and induction time were determined. By adjusting pH and improving ventilation, a fermentation process with high expression of rhAGN was initially formed.