Effect of oxygen, and ArcA and FNR regulators on the expression of genes related to the electron transfer chain and the TCA cycle in Escherichia coli

• Published in: Metabolic engineering Vol. 7; no. 5; pp. 364 - 374
• Main Authors: Shalel-Levanon, Sagit, San, Ka-Yiu, Bennett, George N.
• Format: Journal Article
• Language: English
• Published: Belgium Elsevier Inc 01.09.2005
• Subjects: Bacterial Outer Membrane Proteins - metabolism; Bioreactors - microbiology; Cell Culture Techniques - methods; Citric Acid Cycle - physiology; Cytochrome oxidase; Electron Transport Chain Complex Proteins - metabolism; Escherichia coli - metabolism; Escherichia coli Proteins - metabolism; Gene Expression Regulation, Bacterial - physiology; Iron-Sulfur Proteins - metabolism; Microaerobic conditions; NADH dehydrogenase; Oxygen - metabolism; QRT-PCR; Repressor Proteins - metabolism; TCA cycle; Transcription Factors - metabolism; Cytochrome oxidase; NADH dehydrogenase; Microaerobic conditions; TCA cycle; QRT-PCR
• DOI: 10.1016/j.ymben.2005.07.001
• ISSN: 1096-7176

Microbial cells possess numerous sensing/regulator systems in order to respond rapidly to environmental changes. Escherichia coli has several elaborate sensing mechanisms for response to the availability of oxygen and the presence of other electron acceptors. A group of global regulators, which include the one component Fnr protein and the two-component Arc system, coordinate the adaptive responses. To quantitate the contribution of Arc and FNR-dependent regulation under microaerobic conditions, the gene expression pattern of the electron transfer chain genes and the TCA cycle genes in wild-type E. coli, an arcA mutant, an fnr mutant, and a double arcA, fnr mutant, in glucose limited cultures and different oxygen concentrations was studied in chemostat cultures at steady state using QRT-PCR. It was found that the TCA cycle genes, icd, gltA, sucC, and sdhC are repressed by ArcA while Fnr has a minor or no effect on the expression of these genes under microaerobic conditions. The expression levels of the electron transfer chain genes, nuoA, ndh, and ubiE, were not significantly affected by either ArcA or Fnr regulation proteins, while a lower expression of cydA (up to 9-fold lower) and a higher expression of cyoA (up to 31-fold higher) were observed in cultures of the arcA mutant strain compared to those of the wild type. Since significantly higher NADH/NAD + ratios were previously observed in cultures of the arcA mutant strain compared to the wild type it seems that the cytochrome o oxidase (the product of cyoABCDE) cannot efficiently support aerobic respiration when the cells are grown under microaerobic conditions.