Direct measurement of isotopomer of intracellular metabolites using capillary electrophoresis time-of-flight mass spectrometry for efficient metabolic flux analysis

Yoshihiro Toya, Nobuyoshi Ishii, Takashi Hirasawa, Miki Naba, Kenta Hirai, Kaori Sugawara, Saori Igarashi, Kazuyuki Shimizu, Masaru Tomita, Tomoyoshi Soga

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

We have developed a metabolic flux analysis method that is based on 13C-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.

Original languageEnglish
Pages (from-to)134-141
Number of pages8
JournalJournal of Chromatography A
Volume1159
Issue number1-2
DOIs
Publication statusPublished - 2007 Aug 3

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Metabolic Flux Analysis
Capillary electrophoresis
Capillary Electrophoresis
Metabolites
Mass spectrometry
Mass Spectrometry
Fluxes
Amino Acids
Metabolism
Labeling
Escherichia coli
Hydrolysis
Carbon
Throughput
Proteins

Keywords

  • C-labeling experiment
  • CE-TOFMS
  • Exchange coefficient
  • GC-MS
  • Metabolic flux analysis

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Direct measurement of isotopomer of intracellular metabolites using capillary electrophoresis time-of-flight mass spectrometry for efficient metabolic flux analysis. / Toya, Yoshihiro; Ishii, Nobuyoshi; Hirasawa, Takashi; Naba, Miki; Hirai, Kenta; Sugawara, Kaori; Igarashi, Saori; Shimizu, Kazuyuki; Tomita, Masaru; Soga, Tomoyoshi.

In: Journal of Chromatography A, Vol. 1159, No. 1-2, 03.08.2007, p. 134-141.

Research output: Contribution to journalArticle

Toya, Yoshihiro ; Ishii, Nobuyoshi ; Hirasawa, Takashi ; Naba, Miki ; Hirai, Kenta ; Sugawara, Kaori ; Igarashi, Saori ; Shimizu, Kazuyuki ; Tomita, Masaru ; Soga, Tomoyoshi. / Direct measurement of isotopomer of intracellular metabolites using capillary electrophoresis time-of-flight mass spectrometry for efficient metabolic flux analysis. In: Journal of Chromatography A. 2007 ; Vol. 1159, No. 1-2. pp. 134-141.
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AU - Toya, Yoshihiro

AU - Ishii, Nobuyoshi

AU - Hirasawa, Takashi

AU - Naba, Miki

AU - Hirai, Kenta

AU - Sugawara, Kaori

AU - Igarashi, Saori

AU - Shimizu, Kazuyuki

AU - Tomita, Masaru

AU - Soga, Tomoyoshi

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