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重组人内皮抑素(rhEndostatin)是人内皮抑素经过基因突变获得的新的蛋白质序列,是一种血管生成抑制剂类蛋白,具有抗肿瘤活性。实验对蛋白rhEndostatin产生菌E.coliBL21(DE3)的基本发酵条件进行了研究并用正交试验对培养基成分进行了优化,优化后的培养基组成为:蛋白胨1%,酵母粉0.5%,葡萄糖0.5%,NaCl 0.2%,Na2HPO4.12 H2O 1.5%,KH2PO40.2%,(NH4)2SO40.2%,MgSO4.7 H2O 0.02%,VB15 mg/L,VB21 mg/L,微量元素:CoCl2.6 H2O 5 mg/L,MnCl2.4 H2O,H3BO3,Na2MoO4.2 H2O,AlCl3.6 H2O,Ferric citrate均20 mg/L。优化后的培养基在发酵罐发酵,湿菌重达到45 g/L以上,目的蛋白产量达324 mg/L,为rhEndostatin的进一步研究开发打下了基础。
Recombinant human endostatin (rhEndostatin) is a new protein sequence of human endostatin gene mutation, is an angiogenesis inhibitor protein with anti-tumor activity. The fermentation conditions of rhEndostatin-producing strain E. coli BL21 (DE3) were studied and the components of the medium were optimized by orthogonal test. The optimized medium consisted of peptone 1%, yeast extract 0.5%, glucose 0.5 %, NaCl 0.2%, Na2HPO4.12 H2O 1.5%, KH2PO40.2%, (NH4) 2SO4 40.2%, MgSO4.7 H2O 0.02%, VB15 mg / L, VB21 mg / 5 mg / L, MnCl2.4 H2O, H3BO3, Na2MoO4.2 H2O, AlCl3.6 H2O, and Ferric citrate all 20 mg / L. The optimized culture medium was fermented in a fermenter. The wet weight reached 45 g / L and the target protein yield reached 324 mg / L, which laid the foundation for further research and development of rhEndostatin.