High-throughput metabolic flux analysis based on gas chromatography-mass spectrometry derived 13C constraints

@article{citeulike:3016138, abstract = {13C-constrained flux balancing analysis based on gas chromatography-mass spectrometry data is presented here as a simple and robust method for the estimation of intracellular carbon fluxes. In this approach, the underdetermined system of metabolite balances deduced from stoichiometric relations and measured extracellular rates is complemented with 13C constraints from metabolic flux ratio analysis. Fluxes in central carbon metabolism of exponentially growing Escherichia coli were estimated by 13C-constrained flux balancing from three different 13C-labeled glucose experiments. The best resolution of the network was achieved using 13C constraints derived from [U-13C]glucose and [1-13C]glucose experiments. The corresponding flux estimate was in excellent agreement with a solution that was independently obtained with a comprehensive isotopomer model. This new methodology was also demonstrated to faithfully capture the intracellular flux distribution in E. coli shake flasks and 1-ml deep-well microtiter plates. Due to its simplicity, speed, and robustness, 13C-constrained metabolic flux balancing is promising for routine and high-throughput analysis on a miniaturized scale.}, author = {Fischer, Eliane and Zamboni, Nicola and Sauer, Uwe }, citeulike-article-id = {3016138}, doi = {http://dx.doi.org/10.1016/j.ab.2003.10.036}, journal = {Analytical Biochemistry}, keywords = {flux-measurements, metabolic-engineering, mfa, sauer}, month = {February}, number = {2}, pages = {308--316}, posted-at = {2008-07-18 03:25:48}, priority = {2}, title = {High-throughput metabolic flux analysis based on gas chromatography-mass spectrometry derived 13C constraints}, url = {http://dx.doi.org/10.1016/j.ab.2003.10.036}, volume = {325}, year = {2004} }

TY - JOUR ID - citeulike:3016138 L3 - citeulike-article-id:3016138 TI - High-throughput metabolic flux analysis based on gas chromatography-mass spectrometry derived 13C constraints JF - Analytical Biochemistry VL - 325 IS - 2 SP - 308 EP - 316 N2 - 13C-constrained flux balancing analysis based on gas chromatography-mass spectrometry data is presented here as a simple and robust method for the estimation of intracellular carbon fluxes. In this approach, the underdetermined system of metabolite balances deduced from stoichiometric relations and measured extracellular rates is complemented with 13C constraints from metabolic flux ratio analysis. Fluxes in central carbon metabolism of exponentially growing Escherichia coli were estimated by 13C-constrained flux balancing from three different 13C-labeled glucose experiments. The best resolution of the network was achieved using 13C constraints derived from [U-13C]glucose and [1-13C]glucose experiments. The corresponding flux estimate was in excellent agreement with a solution that was independently obtained with a comprehensive isotopomer model. This new methodology was also demonstrated to faithfully capture the intracellular flux distribution in E. coli shake flasks and 1-ml deep-well microtiter plates. Due to its simplicity, speed, and robustness, 13C-constrained metabolic flux balancing is promising for routine and high-throughput analysis on a miniaturized scale. KW - flux-measurements KW - metabolic-engineering KW - mfa KW - sauer AU - Fischer, Eliane AU - Zamboni, Nicola AU - Sauer, Uwe PY - 2004/02/15/ UR - http://dx.doi.org/10.1016/j.ab.2003.10.036 ER -