論文 - 詳細
| RRC ID | 3229 |
|---|---|
| 著者 | Yang C, Hua Q, Baba T, Mori H, Shimizu K. |
| タイトル | Analysis of Escherichia coli anaplerotic metabolism and its regulation mechanisms from the metabolic responses to altered dilution rates and phosphoenolpyruvate carboxykinase knockout. |
| ジャーナル | Biotechnol Bioeng |
| Abstract |
The gluconeogenic phosphoenolpyruvate (PEP) carboxykinase is active in Escherichia coli during its growth on glucose. The present study investigated the influence of growth rates and PEP carboxykinase knockout on the anaplerotic fluxes in E. coli. The intracellular fluxes were determined using the complementary methods of flux ratio analysis and metabolic flux analysis based on [U-(13)C(6)]glucose labeling experiments and 2D nuclear magnetic resonance (NMR) spectroscopy of cellular amino acids and glycerol. Significant activity of PEP carboxykinase was identified in wild-type E. coli, and the ATP dissipation for the futile cycling via this reaction accounted for up to 8.2% of the total energy flux. Flux analysis of pck deletion mutant revealed that abolishment of PEP carboxykinase activity resulted in a remarkably reduced flux through the anaplerotic PEP carboxylase and the activation of the glyoxylate shunt, with 23% of isocitrate found being channeled in the glyoxylate shunt. The changes in intracellular metabolite concentrations and specific enzyme activities associated with different growth rates and pck deletion, were also determined. Combining the measurement data of in vivo fluxes, metabolite concentrations and enzyme activities, the in vivo regulations of PEP carboxykinase flux, PEP carboxylation, and glyoxylate shunt in E. coli are discussed. |
| 巻・号 | 84(2) |
| ページ | 129-44 |
| 公開日 | 2003-10-20 |
| DOI | 10.1002/bit.10692 |
| PMID | 12966569 |
| MeSH | Acetyl Coenzyme A / metabolism Adenine Nucleotides / metabolism Adenosine Triphosphate / metabolism Algorithms Amino Acids / chemistry Amino Acids / metabolism Carbon / metabolism Carbon Dioxide / metabolism Carbon Isotopes / metabolism Carboxylic Acids / metabolism Cell Division / physiology Citric Acid Cycle Energy Metabolism Escherichia coli / enzymology Escherichia coli / genetics Escherichia coli / metabolism* Gene Deletion Gene Expression Regulation, Bacterial Gene Expression Regulation, Enzymologic Glucose / metabolism Glycolysis Hydrolysis Isocitrate Dehydrogenase / metabolism Isocitrate Lyase / metabolism Kinetics Magnetic Resonance Spectroscopy Models, Biological Monosaccharides / metabolism Mutation Oxygen / metabolism Pentose Phosphate Pathway Phosphoenolpyruvate Carboxykinase (ATP) / deficiency Phosphoenolpyruvate Carboxykinase (ATP) / genetics Phosphoenolpyruvate Carboxykinase (ATP) / metabolism* Phosphoenolpyruvate Carboxylase / metabolism Substrate Cycling |
| IF | 4.002 |
| 引用数 | 95 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| リソース情報 | |
| 原核生物(大腸菌) | W3110 ME9062(BW25113) |