霍乱毒素B亚单位工程菌MM2在含乳酸培养基中的高表达

确定了工程菌MM2中霍乱毒素B亚单位(CTB)在含乳酸培养基中的高表达方案。采用两阶段控制培养温度(30℃→37℃)可提高CTB产量4倍,提高后期pH值(72→84)可提高CTB比表达水平214倍,中间补加乙酸钠可提高CTB产量65%。在5L发酵罐培养菌密度OD₆₀₀达30,CTB产量达1867mg/L,产物在培养液中以多聚体形式存在,具有抗原性。

Overproduction of Cholera Toxin B Subunit by Recombinant Escherichia coli MM2 in Lactate-containing Medium

The expression of ctb gene in recombinant E. coli MM2 strain is affected by temperature, pH and carbon source. High level production of cholera toxin B subunit (CTB) was investigated in lactate-containing medium which designed by experiments of orthogonal test. Upshifting temperature from 30 degrees C to 37 degrees C could increase the production of CTB by 4 fold, upshifting pH value from 7.2 to 8.4 at the later culture stage could increase the specific expression level of CTB by 2.14 fold, and adding sodium acetate increased the production of CTB by 65%. In 5 L fermentor the cell density was reached at OD600 30, and the 186.7 mg/L of CTB was obtained. The CTB in the culture supernatants was in the form of polymer like the native CTB from Sigma, and also possessed the same antigenity.