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首页> 外文期刊>Applied Microbiology >Development of a Novel Plasmid-Free Thymidine Producer by Reprogramming Nucleotide Metabolic Pathways
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Development of a Novel Plasmid-Free Thymidine Producer by Reprogramming Nucleotide Metabolic Pathways

机译：通过重新编程核苷酸代谢途径的新型无质粒胸苷生产者的发展。

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A novel thymidine-producing strain of Escherichia coli was prepared by genome recombineering. Eleven genes were deleted by replacement with an expression cassette, and 7 genes were integrated into the genome. The resulting strain, E. coli HLT013, showed a high thymidine yield with a low deoxyuridine content. DNA microarrays were then used to compare the gene expression profiles of HLT013 and its isogenic parent strain. Based on microarray analysis, the pyr biosynthesis genes and 10 additional genes were selected and then expressed in HLT013 to find reasonable candidates for enhancing thymidine yield. Among these, phage shock protein A (PspA) showed positive effects on thymidine production by diminishing redox stress. Thus, we integrated pspA into the HLT013 genome, resulting in E. coli strain HLT026, which produced 13.2 g/liter thymidine for 120 h with fed-batch fermentation. Here, we also provide a basis for new testable hypotheses regarding the enhancement of thymidine productivity and the attainment of a more complete understanding of nucleotide metabolism in bacteria.
机译：通过基因组重组制备了新的产生胸苷的大肠杆菌菌株。通过替换表达盒删除了11个基因，并将7个基因整合到基因组中。所得菌株大肠杆菌HLT013显示出较高的胸苷产率和较低的脱氧尿苷含量。然后使用DNA微阵列比较HLT013及其同基因亲本菌株的基因表达谱。基于微阵列分析，选择了pyr生物合成基因和另外10个基因，然后在HLT013中表达以寻找提高胸苷产量的合理候选物。其中，噬菌体休克蛋白A（PspA）通过减少氧化还原应激对胸腺嘧啶核苷产生积极影响。因此，我们将pspA整合到HLT013基因组中，产生了大肠杆菌菌株HLT026，该菌株在分批补料发酵的条件下，在120小时内产生了13.2 g /升的胸苷。在这里，我们还为有关可检测的胸腺嘧啶核苷生产率提高和对细菌核苷酸代谢的更完整理解提供了新的可检验假说的基础。

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