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对已构建好的表达HrpNEcc蛋白的工程菌BL21(DE3)/pET30a(+)hrpNEcc的摇瓶发酵条件及乳糖诱导进行优化,通过在7L发酵罐中放大发酵实验,以期提高蛋白产量并降低生产成本。在摇瓶中优化的发酵及诱导条件是:5%的接种量,TB培养基,菌体培养至对数生长前期,添加3g/L外源诱导剂乳糖时,HrpNEcc蛋白产量可达417.60mg/L,比不添加乳糖时提高了36.73%,比用IPTG诱导时提高了16.85%。7L发酵罐中发酵,获得菌体湿重达到57.24g/L(WCW),可溶性HrpNEcc蛋白产量占细胞总蛋白的50.2%,为3.29 g/L。
The shake flask fermentation conditions and lactose induction of the constructed engineering strain BL21 (DE3) / pET30a (+) hrpNEcc expressing HrpNEcc protein were optimized and the fermentation experiment was enlarged in a 7L fermenter in order to increase the protein yield and reduce the production cost . The optimized fermentation and induction conditions in shake flasks were as follows: 5% inoculum volume, TB medium, bacterial culture until logarithmic growth phase, HrpNEcc protein yield up to 417.60mg / L when 3g / L exogenous inducer lactose was added, L, increased by 36.73% when lactose was not added, and increased by 16.85% when induced by IPTG. 7L fermenter. The wet weight of the strain reached 57.24 g / L (WCW), and the soluble HrpNEcc protein production accounted for 50.2% of the total cellular protein, which was 3.29 g / L.