| 大肠杆菌耐热元器件的构建及其应用 |
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| 引用本文： | 孙翔英,刘月芹,孙欢,贾海洋,戴大章,李春.大肠杆菌耐热元器件的构建及其应用[J].化工学报,2014,65(8):3128-3135. |
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| 作者姓名： | 孙翔英 刘月芹 孙欢 贾海洋 戴大章 李春 |
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| 作者单位： | 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 |
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| 基金项目： | 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）。 |
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| 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。
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| 关 键 词： | 耐热性 热激蛋白 存活率 抗逆性 大肠杆菌 |
| 收稿时间： | 2013-11-06 |
| 修稿时间： | 2013-12-24 |
Construction of heat resistance devices for Escherichia coli and their application |
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| SUN Xiangying,LIU Yueqin,SUN Huan,JIA Haiyang,DAI Dazhang,LI Chun.Construction of heat resistance devices for Escherichia coli and their application[J].Journal of Chemical Industry and Engineering(China),2014,65(8):3128-3135. |
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| Authors: | SUN Xiangying LIU Yueqin SUN Huan JIA Haiyang DAI Dazhang LI Chun |
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| Affiliation: | 1.Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan/School of Chemistry and Chemical Engineering, Shihezi University, Shihezi 832003, Xinjiang, China;2.School of Life Science, Beijing Institute of Technology, Beijing 100081, China |
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| Abstract: | 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. |
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| Keywords: | heat resistance heat shock protein (HSP) survival rate stress resistance Escherichia coli |
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