重组人血管抑素生产菌培养基优化的研究

Optimization of Culture Medium for the Production of Recombinant Human Angiostatin

用正交试验对血管抑素生产菌株E .colipQE32 AGN的发酵培养基成分进行了优化 ,优化后的合成培养基组成为 :无水葡萄糖 1% ,(NH4) 2 SO40 .4 % ,NaCl0 .2 % ,KH2 PO40 .3% ,K2 HPO42 % ,MgSO40 .0 1% ,盐酸硫胺素 0 .4mg/L ,核黄素 0 .1mg/L ,盐酸吡哆醇 0 .6mg/L ,D 泛酸 0 .3ml/L ,硫酸亚铁 30mg/L ,硫酸锂 30mg/L ,硫酸锰 2 5mg/L ,醋酸锌 2 0 mg/L ,硫酸铝 9mg/L ,优化后的生物量达到 9.0g/L以上 ,比采用LB培养基培养的生物量提高了一倍多。同时 ,优化培养基对工程菌目的蛋白的表达、溶解及复性未产生不利影响。

An efficient fermentation medium with high yields has been developed for the production of recombinant human angiostatin from pQE32-AGN. Orthogonal Design was applied to optimize the medium constituents. The optimal levels for the constituents were: glucose 1%,(NH 4) 2SO 4 0.4%,NaCl 0.2%,KH 2PO 4 0.3%,K 2HPO 4 2%,Mg SO 4 0.01%,VitaminB 1 0.4 mg/L, VitaminB 2 0.1 mg/L, VitaminB 6 0.6 mg/L, D-Pantothenic acid 0.3 ml/L,FeSO 4 30 mg/L,Li 2SO 4 30 mg/L, MnSO 4 25 mg/L, Zn(CH 3COO) 2 20 mg/L,Al 2(SO 4) 3 9 mg/L. The optimized medium produced 9.0 g/L biomass, which was twice more than LB medium. Moreover, the high-cell density growth of engineering strain has no side effect on the expression, dissolution, renaturation of target protein.