SUN Xiangying, LIU Yueqin, SUN Huan, et al. Construction of heat resistance devices for Escherichia coli and their application[J]. CIESC Journal, 2014, (8): 3128-3135.
SUN Xiangying, LIU Yueqin, SUN Huan, et al. Construction of heat resistance devices for Escherichia coli and their application[J]. CIESC Journal, 2014, (8): 3128-3135.DOI: 10.3969/j.issn.0438-1157.2014.08.038.
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。
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.