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为提高可溶性GST-PLAD蛋白的表达量,对含有GST-PLAD基因的E.coli BL21(DE3)工程菌的发酵条件进行优化。采用单因素实验和正交实验,对影响大肠杆菌生长及融合蛋白表达的培养基组分、培养条件进行了优化。结果显示最优培养基配比(%)为:蔗糖0.5、蛋白胨1、酵母提取物1.5、硫酸铵0.4、氯化钠1、硫酸镁0.03;最佳表达条件为:诱导剂IPTG浓度为0.4 mmol/L,37℃培养5.5 h,诱导9 h,摇瓶装瓶量为100 mL/500 mL。表达条件优化后菌体生长密度是优化前的1.89倍,GST-PLAD表达量是优化前的2.05倍。
To improve the expression of soluble GST-PLAD protein, the fermentation conditions of E. coli BL21 (DE3) engineered bacteria containing the GST-PLAD gene were optimized. Single factor experiments and orthogonal experiments were used to optimize the components of culture medium and culture conditions that affect the growth of Escherichia coli and the expression of fusion protein. The results showed that the optimum medium was as follows: sucrose 0.5, peptone 1, yeast extract 1.5, ammonium sulfate 0.4, sodium chloride 1, magnesium sulfate 0.03. The optimum conditions were as follows: the inducer IPTG concentration was 0.4 mmol / L, cultured at 37 ℃ 5.5 h, induced 9 h, flask bottling 100 mL / 500 mL. After optimization, the cell growth density was 1.89 times that before optimization, and the expression level of GST-PLAD was 2.05 times before optimization.