2P-21 PDF
13C-metabolic flux analysis using isotopomer labeling of intracellular free metabolites
13C metabolic flux analysis (MFA) is a tool of metabolic engineering for investigation of in vivo flux distribution. A direct 13C enrichment analysis of intracellular free amino acids (FAAs) is expected to reduce time for labeling experiments of the MFA. An examination of Escherichia coli showed that the metabolic flux distribution estimated using FAAs has an essentially identical with that of the conventional proteinogenic amino acids-based method, and the 13C enrichment data of glutamate, aspartate, alanine, and phenylalanine is the minimal requirement for a reliable analysis of the flux distribution. It was also suggested that, since perturbation while sampling affects 13C labeling data, fast sampling methods such as cold methanol quenching is essential for MFA using intracellular metabolites.