Chassagnole Christophe, Noisommit-Rizzi Naruemol, Schmid Joachim W, Mauch Klaus, Reuss Matthias
Institute of Biochemical Engineering, University of Stuttgart, Stuttgart, Germany.
Biotechnol Bioeng. 2002 Jul 5;79(1):53-73. doi: 10.1002/bit.10288.
Application of metabolic engineering principles to the rational design of microbial production processes crucially depends on the ability to describe quantitatively the systemic behavior of the central carbon metabolism to redirect carbon fluxes to the product-forming pathways. Despite the importance for several production processes, development of an essential dynamic model for central carbon metabolism of Escherichia coli has been severely hampered by the current lack of kinetic information on the dynamics of the metabolic reactions. Here we present the design and experimental validation of such a dynamic model, which, for the first time, links the sugar transport system (i.e., phosphotransferase system [PTS]) with the reactions of glycolysis and the pentose-phosphate pathway. Experimental observations of intracellular concentrations of metabolites and cometabolites at transient conditions are used to validate the structure of the model and to estimate the kinetic parameters. Further analysis of the detailed characteristics of the system offers the possibility of studying important questions regarding the stability and control of metabolic fluxes.
将代谢工程原理应用于微生物生产过程的合理设计，关键取决于定量描述中心碳代谢系统行为的能力，以便将碳通量重定向至产物形成途径。尽管对多个生产过程都很重要，但由于目前缺乏代谢反应动力学的相关信息，大肠杆菌中心碳代谢基本动态模型的开发受到了严重阻碍。在此，我们展示了这样一个动态模型的设计与实验验证，该模型首次将糖转运系统（即磷酸转移酶系统[PTS]）与糖酵解反应和磷酸戊糖途径联系起来。利用瞬态条件下细胞内代谢物和共代谢物浓度的实验观测结果来验证模型结构并估计动力学参数。对该系统详细特征的进一步分析为研究有关代谢通量稳定性和控制的重要问题提供了可能性。
