| 大肠杆菌L-组氨酸生物合成途径的改造及其对工程菌L-组氨酸产量的影响 |
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| 引用本文： | 魏伟,刘倩,卢利宁,谢希贤. 大肠杆菌L-组氨酸生物合成途径的改造及其对工程菌L-组氨酸产量的影响[J]. 天津科技大学学报, 2011, 0(4): 6-9 |
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| 作者姓名： | 魏伟 刘倩 卢利宁 谢希贤 |
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| 作者单位： | 工业发酵微生物教育部重点实验室天津科技大学生物工程学院; |
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| 基金项目： | 国家“十一五”科技支撑计划项目(2008BAI63B01) |
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| 摘 要： | 改造大肠杆菌L–组氨酸生物合成途径,以提高L–组氨酸的产量.用NTG诱变大肠杆菌M-17(SGr),依次赋予其2–噻唑丙氨酸(2-TA)和组氨酸氧肟酸盐(HisHx)遗传标记,再以突变株M-18(SGr+2-TAr+HisHxr)基因组为模板,扩增组氨酸操纵子基因,构建出重组质粒pUC118-his-operon,将重组质粒导入突变株M-18(SGr+2-TAr+HisHxr)得到工程菌E.coli M-19(SGr+2-TAr+HisHxr/pUC118-his-operon).根据zwf和prs基因序列分别合成引物进行PCR扩增,PCR产物与载体pSTV28连接,构建重组质粒pSTV28-zwf、pSTV28-prs和pSTV28-zwf-prs,将重组质粒分别转化至工程菌E.coli M-19,摇瓶发酵测定重组工程菌L–组氨酸的产量.摇瓶发酵结果显示,L–组氨酸产量与对照株相比,工程菌E.coli M-19提高了4.5倍,双质粒系统重组工程菌E.coli MZH-19、E.coli MPH-19和E.coli MZPH-19分别提高了5.14、5.78、8.43倍.
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| 关 键 词： | L–组氨酸 组氨酸操纵子 大肠杆菌 双质粒系统 摇瓶发酵 |
Modification of L-histidine Biosynthesis Pathway in E. coli and Its Effect on L-histidine Yield |
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| WEI Wei,LIU Qian,LU Li-ning,XIE Xi-xian. Modification of L-histidine Biosynthesis Pathway in E. coli and Its Effect on L-histidine Yield[J]. Journal of Tianjin University of Science & Technology, 2011, 0(4): 6-9 |
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| Authors: | WEI Wei LIU Qian LU Li-ning XIE Xi-xian |
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| Affiliation: | WEI Wei,LIU Qian,LU Li-ning,XIE Xi-xian(Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,College of Biotechnology,Tianjin University of Science & Technology,Tianjin 300457,China) |
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| Abstract: | To modify the metabolic pathway of L-histidine biosynthesis in E.coli and increase L-histidine yield.E.coli M-18(SGr+2-TAr+HisHxr)was obtained by N-Methyl-N-nitro-N-nitrosoguanidine(NTG)mutagenesis derived from the original strain E.coli M-17(SGr).The histidine operon was amplified by PCR from E.coli M-18(SGr + 2-TAr + HisHxr)chromosome and ligated it into the pUC118 vector.The recombinant plasmid pUC118-his-operon was transformed into E.coli M-18(SGr+2-TAr+HisHxr)by electroporation.The zwf gene and prs gen... |
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| Keywords: | L-histidine his operon Escherichia coli two-plasmid system flask-shaking fermentation |
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