基因重组大肠杆菌表达HrpNEcc蛋白的发酵条件及诱导条件优化  

Optimization of Fermentation and Induction Conditions of Recombinant E.coli BL21(DE3)/pET30a(+)hrpNEcc

作　　者：张姝[1,2] 王敏[2] 韩梅琳[2] 马荣才[2] 陈强[1] 高俊莲[2] 

机构地区：[1]四川农业大学资源环境学院微生物学系,雅安625014 [2]北京市农林科学院北京农业生物技术研究中心,北京100097

出　　处：《中国生物工程杂志》2009年第10期44-49,共6页China Biotechnology

基　　金：北京市科学技术委员会农业科技成果转化专项(Z07090500550705);北京市科学技术委员会重大项目专项(Z0005190040531)资助项目

摘　　要：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。In order to obtain high yield of the HrpNEcc protein with a lower total cost,fermentation and lactose induction conditions for recombinant E.coli BL21（DE3）/pET30a（＋）hrpN Ecc were optimized in flasks and the recombinant E.coli was fermentated in 7L fermenter.The optimized incoulum concentration was 5% and the optimized nutrient medium was TB medium.The HrpNEcc protein yield reached 417.60mg/L by adding 3g/L exogenous inducer lactose in the growth prophase of log-phase for the recombinant E.coli.The HrpNEcc protein yield was higher 36.73% than that of the case of no any inducer,and was higher 16.85% than that of the case of adding IPTG.The wet weight of cell pellet of the recombinant E.coli reached 57.24g/L after fermentation in 7L fermenter,the HrpNEcc protein reached 3.29g/L,about 50.2% of total cellular protein.

关 键 词：HrpNEcc蛋白 重组大肠杆菌 发酵及诱导条件优化 

分 类 号：Q93-335[生物学—微生物学]