1887

Abstract

ATCC 17933 growing aerobically on ethanol uses a pyrroloquinoline quinone-dependent ethanol oxidation system. A mutant with an interrupted putative gene, in which malate:quinone oxidoreductase (MQO), an enzyme involved in the citric acid cycle/glyoxylate cycle, was defective, showed a severe growth defect on ethanol and was unable to grow on acetate. Glucose, lactate, succinate or malate supported growth of the mutant. However, an NAD-dependent malate dehydrogenase activity could not be detected. Complementation of the mutant by the wild-type allele of the gene restored wild-type behaviour. The wild-type expressed the dye-dependent MQO and NAD(P)-dependent malic enzymes (MEs). Pyruvate carboxylase (PC) was found upon growth of the wild-type and the mutant on all substrates studied. PC activity in the wild-type was induced on glucose and lactate and was always higher on all substrates in the mutant. In ATCC 17933, an active MQO is required for growth on ethanol or acetate, while with glucose, lactate, succinate or malate an apparent bypass route operates, with MEs using malate for generating pyruvate, which is carboxylated to oxaloacetate by PC. To the authors’ knowledge, this is the first time that a specific mutant MQO phenotype has been observed, caused by the inactivation of a gene encoding MQO activity. of ATCC 17933 corresponds to () of the PAO1 genome project.

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2002-12-01
2021-10-18
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References

  1. Altschul S. F., Madden T. L., Schaffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402 [CrossRef]
    [Google Scholar]
  2. Armstrong J. McD. 1964; The molar extinction coefficient of 2,6-dichlorophenol indophenol. Biochim Biophys Acta 86:194–197 [CrossRef]
    [Google Scholar]
  3. Austin D., Larson T. J. 1991; Nucleotide sequence of the glpD gene encoding aerobic sn -glycerol 3-phosphate dehydrogenase of Escherichia coli K-12. J Bacteriol 173:101–107
    [Google Scholar]
  4. Bergmeyer J., Graßl M. 1987 Methods of Enzymatic Analysis, 3rd edn. Weinheim: VCH;
    [Google Scholar]
  5. Boyer H. W., Roulland-Dussoix D. 1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli . J Mol Biol 14:459–472
    [Google Scholar]
  6. Cetin E. T., Töreci K. I., Ang . 1965; Encapsulated Pseudomonas aeruginosa ( Pseudomonas mucosus ) strains. J Bacteriol 89:1432–1433
    [Google Scholar]
  7. Diehl A., von Wintzingerode F., Görisch H. 1998; Quinoprotein ethanol dehydrogenase of Pseudomonas aeruginosa is a homodimer – sequence of the gene and deduced structural properties of the enzyme. Eur J Biochem 257:409–419 [CrossRef]
    [Google Scholar]
  8. Diesterhaft M. D., Freese E. 1973; Role of pyruvate carboxylase, phosphoenolpyruvate carboxykinase, and malic enzyme during growth and sporulation of Bacillus subtilis . J Biol Chem 248:6062–6070
    [Google Scholar]
  9. Figurski D. H., Helinski D. R. 1979; Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans . Proc Natl Acad Sci USA 76:1648–1652 [CrossRef]
    [Google Scholar]
  10. Geiger O., Görisch H. 1987; Enzymatic determination of pyrroloquinoline quinone using crude membranes from Escherichia coli . Anal Biochem 164:418–423 [CrossRef]
    [Google Scholar]
  11. Groves W. E., Davis F. C. Jr, Sells B. H. 1968; Spectrophotometric determination of microgram quantities of protein without nucleic acid interference. Anal Biochem 22:195–210 [CrossRef]
    [Google Scholar]
  12. Hanahan D. 1983; Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580 [CrossRef]
    [Google Scholar]
  13. Hauck R., Adrian L., Wendler P., Amidjojo M., Hegemann W., Görisch H. 2001; Transformation of 2,2′-dichlorodiisopropyl ether in mixed and pure culture. Appl Microbiol Biotechnol 56:491–495 [CrossRef]
    [Google Scholar]
  14. Hopper D. J., Chapman P. J., Dagley S. 1970; Metabolism of l-malate and d-malate by a species of Pseudomonas . J Bacteriol 104:1197–1202
    [Google Scholar]
  15. Horecker B. L., Kornberg A. 1948; The extinction coefficients of the reduced band of pyridine nucleotides. J Biol Chem 175:385–390
    [Google Scholar]
  16. Kather B., Stingl K., van der Rest M. E., Altendorf K., Molenaar D. 2000; Another unusual type of citric acid cycle enzyme in Helicobacter pylori : the malate: quinone oxidoreductase. J Bacteriol 182:3204–3209 [CrossRef]
    [Google Scholar]
  17. Kretzschmar U., Schobert M., Görisch H. 2001; The Pseudomonas aeruginosa acsA gene, encoding an acetyl-CoA synthetase, is essential for growth on ethanol. Microbiology 147:2671–2677
    [Google Scholar]
  18. Leighton M. P., Kelly D. J., Williamson M. P., Shaw J. G. 2001; An NMR and enzyme study of the carbon metabolism of Neisseria meningitidis . Microbiology 147:1473–1482
    [Google Scholar]
  19. Mizuno T., Kageyama M. 1978; Separation and characterization of the outer membrane of Pseudomonas aeruginosa . J Biochem 84:179–191
    [Google Scholar]
  20. Molenaar D., van der Rest M. E., Petroviæ S. 1998; Biochemical and genetic characterization of the membrane-associated malate dehydrogenase (acceptor) from Corynebacterium glutamicum . Eur J Biochem 254:395–403 [CrossRef]
    [Google Scholar]
  21. Molenaar D., van der Rest M. E., Drysch A., Yücel R. 2000; Functions of the membrane-associated and cytoplasmic malate dehydrogenases in the citric acid cycle of Corynebacterium glutamicum . J Bacteriol 182:6884–6891 [CrossRef]
    [Google Scholar]
  22. O’Brien R. W., Taylor B. L. 1977; Formation and dissimilation of oxaloacetate and pyruvate in Pseudomonas citronellolis grown on noncarbohydrate substrates. J Bacteriol 130:131–135
    [Google Scholar]
  23. O’Brien R. W., Chuang D. T., Taylor B. L., Utter M. F. 1977; Novel enzymic machinery for the metabolism of oxaloacetate, phosphoenolpyruvate, and pyruvate in Pseudomonas citronellolis . J Biol Chem 252:1257–1263
    [Google Scholar]
  24. Peters-Wendisch P. G., Wendisch V. F., Paul S., Eikmanns B. J., Sahm H. 1997; Pyruvate carboxylase as an anaplerotic enzyme in Corynebacterium glutamicum . Microbiology 143:1095–1103 [CrossRef]
    [Google Scholar]
  25. Reichmann P., Görisch H. 1993; Cytochrome c 550 from Pseudomonas aeruginosa . Biochem J 289:173–178
    [Google Scholar]
  26. Roehl R. A., Feary T. W., Phibbs P. V. Jr 1983; Clustering of mutations affecting central pathway enzymes of carbohydrate catabolism in Pseudomonas aeruginosa . J Bacteriol 156:1123–1129
    [Google Scholar]
  27. Rothmel R. K., Chakrabarty A. M., Berry A., Darzins A. 1991; Genetic systems in Pseudomonas . Methods Enzymol 204:485–514
    [Google Scholar]
  28. Rupp M., Görisch H. 1988; Purification, crystallization and characterization of quinoprotein ethanol dehydrogenase from Pseudomonas aeruginosa . Biol Chem Hoppe Seyler 369:431–439 [CrossRef]
    [Google Scholar]
  29. Saeki A., Matsushita K., Takeno S., Taniguchi M., Toyama H., Theeragool G., Lotong N., Adachi O. 1999; Enzymes responsible for acetate oxidation by acetic acid bacteria. Biosci Biotechnol Biochem 63:2102–2109 [CrossRef]
    [Google Scholar]
  30. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  31. Schobert M., Görisch H. 1999; Cytochrome c 550 is an essential component of the quinoprotein ethanol oxidation system in Pseudomonas aeruginosa : cloning and sequencing of the genes encoding cytochrome c 550 and an adjacent acetaldehyde dehydrogenase. Microbiology 145:471–481 [CrossRef]
    [Google Scholar]
  32. Schobert M., Görisch H. 2001; A soluble two-component regulatory system controls expression of quinoprotein ethanol dehydrogenase (QEDH) but not expression of cytochrome c 550 of the ethanol-oxidation system in Pseudomonas aeruginosa . Microbiology 147:363–372
    [Google Scholar]
  33. Schweizer H. P., Klassen T. R., Hoang T. 1996; Improved methods for gene analysis in Pseudomonas . In Molecular Biology of Pseudomonads pp 229–237 Edited by Nakazawa T., Furukawa K., Haas D., Silver S. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  34. Simon R., O’Connell M., Labes M., Pühler A. 1986; Plasmid vectors for the genetic analysis and manipulation of rhizobia and other gram-negative bacteria. Methods Enzymol 118:640–659
    [Google Scholar]
  35. Smith A. W., Iglewski B. H. 1989; Transformation of Pseudomonas aeruginosa by electroporation. Nucleic Acids Res 17:10509 [CrossRef]
    [Google Scholar]
  36. Staskawicz B., Dahlbeck D., Keen N., Napoli C. 1987; Molecular characterization of cloned avirulence genes from race 0 to race 1 of Pseudomonas syringae pv. glycinea . J Bacteriol 169:5789–5794
    [Google Scholar]
  37. van der Rest M. E., Frank C., Molenaar D. 2000; Functions of the membrane-associated and cytoplasmic malate dehydrogenases in the citric acid cycle of Escherichia coli . J Bacteriol 182:6892–6899 [CrossRef]
    [Google Scholar]
  38. Voegele R. T., Mitsch M. J., Finan T. M. 1999; Characterization of two members of a novel malic enzyme class. Biochim Biophys Acta 1432:275–285 [CrossRef]
    [Google Scholar]
  39. Wendisch V. F., de Graaf A. A., Sahm H., Eikmann B. J. 2000; Quantitative determination of metabolic fluxes during coutilization of two carbon sources: comparative analyses with Corynebacterium glutamicum during growth on acetate and/or glucose. J Bacteriol 182:3088–3096 [CrossRef]
    [Google Scholar]
  40. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13mp18 and pUC19 vectors. Gene 33:103–119 [CrossRef]
    [Google Scholar]
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