Direct measurement of isotopomer of intracellular metabolites using capillary electrophoresis time-of-flight mass spectrometry for efficient metabolic flux analysis.
We have developed a metabolic flux analysis method that is based on (13)C-labeling patterns of the intracellular metabolites directly measured by capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS). The flux distribution of the central carbon metabolism in Escherichia coli was determined by this new approach and the results were compared with findings obtained by conventional GC-MS analysis based on isotopomer of the proteinogenic amino acids. There were some differences in estimation results between new approach using CE-TOFMS and conventional approach using GC-MS. These were thought to be attributable to variations in measured mass distributions between amino acids and the corresponding precursors and to differences in the sensitivity of the exchange coefficients to mass distributions. However, our CE-TOFMS method facilitates high-throughput flux analysis without requiring complicated sample preparation such as hydrolysis of proteins and derivatization of amino acids.
Toya
Yoshihiro
Y
Institute for Advanced Biosciences, Keio University, 403-1 Nipponkoku, Daihoji, Tsuruoka, Yamagata 997-0017, Japan.
Ishii
Nobuyoshi
N
Hirasawa
Takashi
T
Naba
Miki
M
Hirai
Kenta
K
Sugawara
Kaori
K
Igarashi
Saori
S
Shimizu
Kazuyuki
K
Tomita
Masaru
M
Soga
Tomoyoshi
T
D053898:Biosynthetic Pathways; D002244:Carbon / Q000032:analysis / Q000378:metabolism*; D002247:Carbon Isotopes / Q000378:metabolism*; D003627:Data Interpretation, Statistical; D019075:Electrophoresis, Capillary; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D008401:Gas Chromatography-Mass Spectrometry; D007553:Isotope Labeling; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D002247:Carbon Isotopes; D029968:Escherichia coli Proteins; D002244:Carbon