Hollinshead Whitney, He Lian, Tang Yinjie J
Department of Energy, Environmental and Chemical Engineering, Washington University, St. Louis, MO, USA.
Methods Mol Biol. 2019;1927:215-230. doi: 10.1007/978-1-4939-9142-6_15.
C-assisted metabolism analysis provides rigorous calculations of the intracellular reaction rates (i.e., fluxes) within the central metabolism of microbial hosts. This mapping of the intracellular network within microbes has proven to be essential for understanding the cell physiology. The approach is also a key to identifying central metabolic nodes, probing the rigidity of a metabolic network, revealing cofactor balances, and delineating hidden pathways. Here we present the methodology of using stable isotopic carbon substrates for both qualitative (C-fingerprinting of functional pathways) and quantitative (Metabolic Flux Analysis) metabolism studies on bacterial species. In this methodology, we include step-by-step instructions to use the open source WUflux software for the steady-state flux calculations based on labeling information of amino acids or free metabolites.
碳辅助代谢分析可对微生物宿主中心代谢内的细胞内反应速率（即通量）进行精确计算。事实证明，这种对微生物细胞内网络的映射对于理解细胞生理学至关重要。该方法也是识别中心代谢节点、探究代谢网络的刚性、揭示辅因子平衡以及描绘隐藏途径的关键。在此，我们介绍使用稳定同位素碳底物进行细菌物种定性（功能途径的碳指纹分析）和定量（代谢通量分析）代谢研究的方法。在该方法中，我们提供了逐步说明，以使用开源的WUflux软件根据氨基酸或游离代谢物的标记信息进行稳态通量计算。
