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用均匀设计法对重组人干扰素α-2b基因工程菌的摇瓶发酵条件进行了优化,确定摇瓶培养基组成为:每升含蛋白胨10 g、葡萄糖6 g、酵母粉5 g、Na2HPO4 6 g、KH2PO4 2 g、NH4Cl 0.8 g、NaCl 4 g、CaCl2 0.01 g、MgSO4 0.2 g,发酵表达条件为:pH 6.9,温度37℃,IPTG诱导后表达时间为5 h左右,装料量为20%。三批小试结果为:湿菌体平均产率18 g/L,生物活性3.93×108 IU/L。发酵罐三批中试放大试验,湿菌体平均产率为29 g/L,生物活性为6.68×108 IU/L发酵液。
The fermentation conditions of recombinant human interferon α-2b genetically engineered bacteria were optimized by uniform design. The shake flask culture medium was determined to be composed of 10 g per liter of peptone, 6 g of glucose, 5 g of yeast powder, 5 g of Na2HPO4 6 g, KH2PO4 2 g, NH4Cl 0.8 g, NaCl 4 g, CaCl2 0.01 g, MgSO4 0.2 g. The fermentation conditions were as follows: pH 6.9, temperature 37 ℃, IPTG induction time about 5 h, loading capacity 20% . Three batches of small test results: the average wet cell yield 18 g / L, bioactivity 3.93 × 108 IU / L. Fermentor three batches of pilot scale-up test, the wet cell average yield of 29 g / L, the biological activity of 6.68 × 108 IU / L fermentation broth.