| 1,3-丙二醇氧化还原酶同功酶基因yqhD在肺炎克雷伯氏菌中的表达研究 |
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| 引用本文： | 朱建国,李霜,纪晓俊,胡南,黄和.1,3-丙二醇氧化还原酶同功酶基因yqhD在肺炎克雷伯氏菌中的表达研究[J].食品与发酵工业,2008,34(8). |
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| 作者姓名： | 朱建国 李霜 纪晓俊 胡南 黄和 |
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| 作者单位： | 1. 南京工业大学制药与生命科学学院,江苏,南京,210009
2. 南京工业大学制药与生命科学学院,江苏,南京,210009;南京工业大学江苏省工业生物技术创新中心,江苏,南京,211816 |
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| 基金项目： | 国家重点基础研究发展计划(973计划)，国家高技术研究发展计划(863计划) |
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| 摘 要： | 以大肠杆菌(Escherichia coli)基因组为模板,PCR扩增出1,3-丙二醇氧化还原酶同功酶基因yqhD,经测序确认,将yqhD基因与四环素抗性基因TetR同时插入表达载体pUC18构建重组质粒pUC18-yqhD-TetR,重组质粒转化Klebsiella pneumoniae ME308;37℃,经1.0mmol/LIPTG诱导8h,重组肺炎克雷伯氏菌中1,3-丙二醇氧化还原酶同功酶的酶活力达到4.16U/mg,而对照菌株的酶活仅为0.62U/mg;将重组菌在摇瓶中进行微氧发酵培养,经IPTG诱导,可将60g/L甘油转化为33.8g/L1,3-丙二醇。
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| 关 键 词： | 1 3-丙二醇氧化还原酶同功酶 肺炎克雷伯氏菌 1 3-丙二醇 电转化 |
Expression of Gene yqhD Encoding 1,3-peopanediol Oxidoreductase Isoenzyme in Klebsiella pnemoniae |
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| Zhu Jianguo,Li Shuang,Ji Xiaojun,Hu Nan,Huang He.Expression of Gene yqhD Encoding 1,3-peopanediol Oxidoreductase Isoenzyme in Klebsiella pnemoniae[J].Food and Fermentation Industries,2008,34(8). |
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| Authors: | Zhu Jianguo Li Shuang Ji Xiaojun Hu Nan Huang He |
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| Abstract: | The gene yqhD from Escherichia coli encoding 1,3-propanediol oxidoreductase isoenzyme was amplified by PCR.After combining yqhD and the tetracycline resistant gene TetR,they were connected to plasmid pUC18 to yield the recombinant plasimid pUC18-yqhD-TetR.The recombinant plasmid was transformed into E.coli DH5α and Klebsiella pneumoniae ME308 separately and the 1,3-propanediol oxidoreductase isoenzyme was successfully expressed.The 1,3-propanediol oxidoreductase isoenzyme of recombinant Klebsiella pneumoniae reached 4.16 U/mg under the induction of IPTG at 37 ℃ for 8 h,while at the same condition the enzyme activity of the original strain was only 0.62 U/mg.In the fermentation process of recombinant Klebsiella pneumoniae,60 g/L glycerol was transformed into 33.8 g/L 1,3-propanediol. |
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| Keywords: | 1 3-proapnediol oxidoreductase isoenzyme Klebsiella pneumoniae 1 3-propanediol electroporation |
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