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 language | English |
|---|---|
| Pages (from-to) | 134-141 |
| Number of pages | 8 |
| Journal | Journal of Chromatography A |
| Volume | 1159 |
| Issue number | 1-2 |
| DOIs | |
| Publication status | Published - 2007 Aug 3 |
Fingerprint
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 journal › Article
}
TY - JOUR
T1 - Direct measurement of isotopomer of intracellular metabolites using capillary electrophoresis time-of-flight mass spectrometry for efficient metabolic flux analysis
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
PY - 2007/8/3
Y1 - 2007/8/3
N2 - 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.
AB - 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.
KW - C-labeling experiment
KW - CE-TOFMS
KW - Exchange coefficient
KW - GC-MS
KW - Metabolic flux analysis
UR - http://www.scopus.com/inward/record.url?scp=34447276204&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34447276204&partnerID=8YFLogxK
U2 - 10.1016/j.chroma.2007.04.011
DO - 10.1016/j.chroma.2007.04.011
M3 - Article
C2 - 17462663
AN - SCOPUS:34447276204
VL - 1159
SP - 134
EP - 141
JO - Journal of Chromatography
JF - Journal of Chromatography
SN - 0021-9673
IS - 1-2
ER -