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Effect of redox imbalance on the fitness of Escherichia coli strains engineered for production of biofuels and biochemicals.

机译：氧化还原失衡对设计用于生产生物燃料和生化物质的大肠杆菌菌株适应性的影响。

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摘要

Bacterial metabolism is a set of inter-connnected reactions in the cell upon which a substrate molecule is converted to another molecule and the energy generated is utilized for the multiplication of the organism. For a bacterial fermentation process to function, the number of electron between the substrate and the desired product need to be balanced. Redox balancing is thus an important consideration in metabolic engineering and strain development as well as fermentation design and control. The internal redox state of the cell is known to affect a broad range of genes and cellular functions, metabolite profiles, and membrane transport and other key functions. In our first study, we observed that the redox state of the cell also affects another key cellular function: genetic stability. By measuring the mutation rate in different E. coli clones, we observed a correlation between correlation between the intracellular redox ratio (NADH/NAD+) and the mutagenesis rate of the organism. Further studies revealed that this increase in mutation was caused by the stress induced downregulation of the DNA repair. In our next study, we sought to expand our toolbox for the redox engineering by identifying a set of gene targets whose activity could be modulated to induce changes in the intracellular redox ratio. We next applied our understanding to improve the fitness of the redox-impaired strains of Escherichia coli engineered for the production of succinic acid. These efforts lead to the creation of a strain capable of producing 60% higher succinate than previous reports.
机译：细菌代谢是细胞中一系列相互连接的反应，在该反应上底物分子被转化为另一种分子，并且所产生的能量被用于生物体的繁殖。为了使细菌发酵过程起作用，需要平衡底物和所需产物之间的电子数量。因此，氧化还原平衡是代谢工程和菌株开发以及发酵设计和控制中的重要考虑因素。已知细胞的内部氧化还原状态会影响广泛的基因和细胞功能，代谢产物谱以及膜转运和其他关键功能。在我们的第一项研究中，我们观察到细胞的氧化还原状态还影响另一个关键的细胞功能：遗传稳定性。通过测量不同大肠杆菌克隆中的突变率，我们观察到细胞内氧化还原比（NADH / NAD +）与生物体诱变率之间的相关性。进一步的研究表明，这种突变的增加是由压力诱导的DNA修复的下调引起的。在我们的下一个研究中，我们试图通过鉴定一组基因靶标来扩展氧化还原工程的工具箱，这些靶标的活性可以被调节以诱导细胞内氧化还原比的变化。接下来，我们运用我们的理解来改善为生产琥珀酸而设计的大肠杆菌的氧化还原受损菌株的适应性。这些努力导致产生了一种菌株，该菌株能够产生比先前报道高出60％的琥珀酸盐。

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  • 作者

    Singh, Amarjeet.;

  • 作者单位

    University of Colorado at Boulder.;

  • 授予单位 University of Colorado at Boulder.;
  • 学科 Biology Microbiology.;Engineering Chemical.
  • 学位 Ph.D.
  • 年度 2010
  • 页码 147 p.
  • 总页数 147
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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