大肠杆菌耐热元器件的构建及其应用  

Construction of heat resistance devices for Escherichia coli and their application

作　　者：孙翔英 刘月芹[2] 孙欢[2] 贾海洋[2] 戴大章[2] 李春[1,2] 

机构地区：[1]石河子大学化学化工学院/兵团绿色化工过程重点实验室,新疆石河子832003 [2]北京理工大学生命学院生物工程系,北京100081

出　　处：《化工学报》2014年第8期3128-3135,共8页CIESC Journal

基　　金：国家重点基础研究发展计划项目(2013CB733900);国家自然科学基金项目(21376028);博士点基金项目(20121101110050)~~

摘　　要：以提高大肠杆菌耐热性为目的，基于腾冲嗜热菌（Thermoanaerobacter tengcongensis MB4）热激蛋白基因T.te-HSP20构建了诱导型耐热元器件 T7-T.te-HSP20和组成型耐热元器件 gapA-T.te-HSP20，转入大肠杆菌（Escherichia coli）获得工程菌 E. coli-TH 和 E. coli-GH。工程菌E. coli-TH在30℃和IPTG诱导下，目标蛋白呈可溶性表达，经50℃热激30 min后，存活率提高了3.2倍。高温发酵表明gapA-T.te-HSP20扩宽了工程菌E. coli-GH的最适生长温度的范围（37～43℃），较大程度提高了大肠杆菌的耐热性。抗逆性分析还发现工程菌E. coli-GH具备了耐热与耐丁醇的双重功能，并有一定的抗乙酸和乙醇能力。为工业梯度升温发酵生产生物基产品的高效制造、节省成本提供了新思路。To improve the heat resistance of Escherichia coli, an inducible heat-resistance device T7-T.te-HSP20 and a constitutive heat-resistance device gapA-T.te-HSP20 based on T.te-HSP20 gene from Thermoanaerobacter tengcongensis MB4, and corresponding engineered strains E. coli-TH and E. coli-GH were constructed. The targeted protein was expressed in solubility after IPTG induction at 30℃ in E. coli-TH. Meanwhile, the survival rate of E. coli-TH was 3.2 times higher than the control at 50℃ for 30 min. The result of high-temperature fermentation showed that the optimum temperature range of E. coli-GH was broadened （37-43℃） under the regulation of heat resistance device gapA-T.te-HSP20. Stress resistance analysis showed that E. coli-GH not only possessed heat resistance and butanol resistance, but also had some resistance to acetic acid and ethanol. These results provide a new idea for modern microorganisms industry.

关 键 词：耐热性 热激蛋白 存活率 抗逆性 大肠杆菌 

分 类 号：Q789[生物学—分子生物学]