title: Pyruvate dehydrogenase complex regulator (PdhR) gene deletion boosts glucose metabolism in Escherichia coli under oxygen-limited culture conditions.
abstract: Pyruvate dehydrogenase complex regulator (PdhR) is a transcriptional regulator that negatively regulates formation of pyruvate dehydrogenase complex (PDHc), NADH dehydrogenase (NDH)-2, and cytochrome bo3oxidase in Escherichia coli. To investigate the effects of a PdhR defect on glucose metabolism, a pdhR deletion mutant was derived from the wild-type E. coli W1485 strain by λ Red-mediated recombination. While no difference in the fermentation profiles was observed between the two strains under oxygen-sufficient conditions, under oxygen-limited conditions, the growth level of the wild-type strain was significantly decreased with retarded glucose consumption accompanied by by-production of substantial amounts of pyruvic acid and acetic acid. In contrast, the mutant grew and consumed glucose more efficiently than did the wild-type strain with enhanced respiration, little by-production of pyruvic acid, less production yield and rates of acetic acid, thus displaying robust metabolic activity. As expected, increased activities of PDHc and NDH-2 were observed in the mutant. The increased activity of PDHc may explain the loss of pyruvic acid by-production, probably leading to decreased acetic acid formation, and the increased activity of NDH-2 may explain the enhanced respiration. Measurement of the intracellular NAD+/NADH ratio in the mutant revealed more oxidative or more reductive intracellular environments than those in the wild-type strain under oxygen-sufficient and -limited conditions, respectively, suggesting another role of PdhR: maintaining redox balance in E. coli. The overall results demonstrate the biotechnological advantages of pdhR deletion in boosting glucose metabolism and also improve our understanding of the role of PdhR in bacterial physiology.
authors.affiliation: Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan. Electronic address: mdsy1220@frontier.hokudai.ac.jp.
authors.forename: Soya
authors.initials: S
authors.lastname: Maeda
authors.affiliation: Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan. Electronic address: ku-shimizu@meg-snow.com.
authors.forename: Kumiko
authors.initials: K
authors.lastname: Shimizu
authors.affiliation: Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan. Electronic address: blue_basil502@hotmail.com.
authors.forename: Chie
authors.initials: C
authors.lastname: Kihira
authors.affiliation: Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan. Electronic address: dun.rim01@gmail.com.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Iwabu
authors.affiliation: Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan. Electronic address: rk11eco@gmail.com.
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Kato
authors.affiliation: Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan. Electronic address: amefrasi.mako@gmail.com.
authors.forename: Makoto
authors.initials: M
authors.lastname: Sugimoto
authors.affiliation: Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan. Electronic address: s-fukiya@chem.agr.hokudai.ac.jp.
authors.forename: Satoru
authors.initials: S
authors.lastname: Fukiya
authors.affiliation: Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan. Electronic address: wada@chem.agr.hokudai.ac.jp.
authors.forename: Masaru
authors.initials: M
authors.lastname: Wada
authors.affiliation: Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan. Electronic address: yokota@chem.agr.hokudai.ac.jp.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Yokota
mesh_terms: D019342:Acetic Acid / Q000378:metabolism; D019069:Cell Respiration; D003573:Cytochrome b Group; D003580:Cytochromes / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005285:Fermentation / Q000187:drug effects; D017353:Gene Deletion; D005809:Genes, Regulator / Q000235:genetics; D005947:Glucose / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D009245:NADH Dehydrogenase / Q000378:metabolism; D010084:Oxidation-Reduction / Q000187:drug effects; D010100:Oxygen / Q000378:metabolism* / Q000494:pharmacology; D011768:Pyruvate Dehydrogenase Complex / Q000096:biosynthesis / Q000378:metabolism*; D019289:Pyruvic Acid / Q000378:metabolism; D012097:Repressor Proteins / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism
chemical_list: D003573:Cytochrome b Group; D003580:Cytochromes; D029968:Escherichia coli Proteins; C084955:PdhR protein, E coli; D011768:Pyruvate Dehydrogenase Complex; D012097:Repressor Proteins; C083850:cytochrome bo3, E coli; D009243:NAD; D019289:Pyruvic Acid; D009245:NADH Dehydrogenase; D005947:Glucose; D019342:Acetic Acid; D010100:Oxygen
keywords: Acetic acid; NAD(+)/NADH ratio; NADH dehydrogenase-2; PdhR; Pyruvate dehydrogenase complex; Redox balance