A convenient NMR method for in situ observation of aerobically cultured cells.
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.
Takesada
H
H
Central Research Laboratories, Ajinomoto Co. Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, Kanagawa 210-8681, Japan.
Ebisawa
K
K
Toyosaki
H
H
Suzuki
E I
EI
Kawahara
Y
Y
Kojima
H
H
Tanaka
T
T
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
D003470:Culture Media; D000255:Adenosine Triphosphate