Metabolic regulation analysis of wild-type and arcA mutant Escherichia coli under nitrate conditions using different levels of omics data.
It is of practical interest to investigate the effect of nitrates on bacterial metabolic regulation of both fermentation and energy generation, as compared to aerobic and anaerobic growth without nitrates. Although gene level regulation has previously been studied for nitrate assimilation, it is important to understand this metabolic regulation in terms of global regulators. In the present study, therefore, we measured gene expression using DNA microarrays, intracellular metabolite concentrations using CE-TOFMS, and metabolic fluxes using the (13)C-labeling technique for wild-type E. coli and the ΔarcA (a global regulatory gene for anoxic response control, ArcA) mutant to compare the metabolic state under nitrate conditions to that under aerobic and anaerobic conditions without nitrates in continuous culture conditions at a dilution rate of 0.2 h(-1). In wild-type, although the measured metabolite concentrations changed very little among the three culture conditions, the TCA cycle and the pentose phosphate pathway fluxes were significantly different under each condition. These results suggested that the ATP production rate was 29% higher under nitrate conditions than that under anaerobic conditions, whereas the ATP production rate was 10% lower than that under aerobic conditions. The flux changes in the TCA cycle were caused by changes in control at the gene expression level. In ΔarcA mutant, the TCA cycle flux was significantly increased (4.4 times higher than that of the wild type) under nitrate conditions. Similarly, the intracellular ATP/ADP ratio increased approximately two-fold compared to that of the wild-type strain.
Institute for Advanced Biosciences, Keio University, Tsuruoka 997-0017, Japan. ytoya@sfc.keio.ac.jp
Yoshihiro
Y
Toya
Kenji
K
Nakahigashi
Masaru
M
Tomita
Kazuyuki
K
Shimizu
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000096:biosynthesis; D000332:Aerobiosis; D000693:Anaerobiosis; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D002247:Carbon Isotopes; D002952:Citric Acid Cycle / Q000187:drug effects* / Q000235:genetics; D003470:Culture Media; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D055432:Metabolomics; D009566:Nitrates / Q000494:pharmacology*; D020411:Oligonucleotide Array Sequence Analysis; D010100:Oxygen / Q000378:metabolism; D010427:Pentose Phosphate Pathway / Q000187:drug effects* / Q000235:genetics; D012097:Repressor Proteins / Q000172:deficiency / Q000235:genetics*
D001425:Bacterial Outer Membrane Proteins; D002247:Carbon Isotopes; D003470:Culture Media; D029968:Escherichia coli Proteins; D009566:Nitrates; D012097:Repressor Proteins; C041610:arcA protein, E coli; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D010100:Oxygen