title: Precise metabolic flux analysis of coryneform bacteria by gas chromatography-mass spectrometry and verification by nuclear magnetic resonance.
abstract: Precise metabolic flux analysis (MFA) by gas chromatography-mass spectrometry (GC-MS) and computer calculation was performed, and the consistency of the estimated results was verified by independently performed nuclear magnetic resonance (NMR) analysis. The precise estimation of flux by the integration method of the mass isotopomer signal, defined as the coefficient of variance (CV) of multiple determination, was investigated, and the results estimated using different data sets with the same magnitude of error were confirmed. The CV of multiple determinations was sufficiently small to discuss and compare the fluxes of a metabolic pathway. The estimated fluxes using the GC-MS data were cross-validated with the NMR data that were independently measured and not used for MFA. The developed method was successfully applied to the MFA of the growth phase of two different glutamate-producing coryneform bacteria, Corynebacterium glutamicum and C. efficiens. The difference in the growth rate between these two bacterial species was discussed while considering the results of MFA, including forward and backward (exchange) fluxes.
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Shirai
authors.forename: Kunihiro
authors.initials: K
authors.lastname: Matsuzaki
authors.forename: Masanori
authors.initials: M
authors.lastname: Kuzumoto
authors.forename: Keisuke
authors.initials: K
authors.lastname: Nagahisa
authors.forename: Chikara
authors.initials: C
authors.lastname: Furusawa
authors.forename: Suteaki
authors.initials: S
authors.lastname: Shioya
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shimizu
mesh_terms: D000192:Actinomycetales / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy; D009249:NADP / Q000378:metabolism
chemical_list: D009249:NADP