title: A convenient NMR method for in situ observation of aerobically cultured cells.
abstract: We observed P31-NMR signals of intracellular phosphorylated metabolites, i.e. ATP, NAD(H) and UDPG, in the aerobically cultured cells of AJ13375 (a derivative of E. coli K-12) without a cell culture circulating system, which needs much more cells. Each NMR spectrum of 40 ml cell-culture was obtained with a 25 mm Phi sample tube mini-fermenter equipped with a pH electrode and three supply routes for O2 gas, aqueous ammonia and glucose. Spectra were measured at 15-min intervals. When glucose in the culture medium was consumed, P31-NMR signals of ATP disappeared first and then those of ADP decreased to below the limit of detection. The intracellular concentration of ATP was estimated to be approximately 7 mM.
authors.affiliation: Central Research Laboratories, Ajinomoto Co. Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, Kanagawa 210-8681, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Takesada
authors.forename: K
authors.initials: K
authors.lastname: Ebisawa
authors.forename: H
authors.initials: H
authors.lastname: Toyosaki
authors.forename: E I
authors.initials: EI
authors.lastname: Suzuki
authors.forename: Y
authors.initials: Y
authors.lastname: Kawahara
authors.forename: H
authors.initials: H
authors.lastname: Kojima
authors.forename: T
authors.initials: T
authors.lastname: Tanaka
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000332:Aerobiosis; D003470:Culture Media / Q000378:metabolism; D004734:Energy Metabolism; D004926:Escherichia coli / Q000254:growth & development* / Q000378:metabolism* / Q000502:physiology; D005285:Fermentation / Q000502:physiology; D007424:Intracellular Fluid / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D010766:Phosphorylation; D013997:Time Factors
chemical_list: D003470:Culture Media; D000255:Adenosine Triphosphate