Construction of heat resistance devices for Escherichia coli and their application

doi:10.3969/j.issn.0438-1157.2014.08.038

CIESC Journal ›› 2014, Vol. 65 ›› Issue (8): 3128-3135.DOI: 10.3969/j.issn.0438-1157.2014.08.038

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Construction of heat resistance devices for Escherichia coli and their application

SUN Xiangying¹, LIU Yueqin², SUN Huan², JIA Haiyang², DAI Dazhang², LI Chun^1,2

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

Received:2013-11-06 Revised:2013-12-24 Online:2014-08-05 Published:2014-08-05
Supported by:
supported by the National Basic Research Program of China (2013CB733900), the National Natural Science Foundation of China (21376028) and the Doctoral Fund of Ministry of Education of China (20121101110050).

大肠杆菌耐热元器件的构建及其应用

孙翔英¹, 刘月芹², 孙欢², 贾海洋², 戴大章², 李春^1,2

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

通讯作者: 李春
基金资助:
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）。

Abstract

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.

Key words: heat resistance, heat shock protein (HSP), survival rate, stress resistance, Escherichia coli

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。

关键词: 耐热性, 热激蛋白, 存活率, 抗逆性, 大肠杆菌

CLC Number:

Q789

SUN Xiangying, LIU Yueqin, SUN Huan, JIA Haiyang, DAI Dazhang, LI Chun. Construction of heat resistance devices for Escherichia coli and their application[J]. CIESC Journal, 2014, 65(8): 3128-3135.

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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Construction of heat resistance devices for Escherichia coli and their application

大肠杆菌耐热元器件的构建及其应用

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