1887

Abstract

ATCC 17933 uses a pyrroloquinoline quinone-dependent ethanol oxidation system. Two mutants of , unable to grow on ethanol and showing no acetyl-CoA synthetase (ACS) activity under standard test conditions, were complemented by cosmid pTB3018. Subcloning led to the isolation of a gene which encodes a protein with high similarity to acetyl-CoA synthetases. Interruption of the putative gene by a kanamycin-resistance cassette resulted in a mutant also unable to grow on ethanol and with very low residual acetyl-CoA-forming activity. Complementation by the wild-type allele of the gene restored growth and led to the expression of ACS activity in excess of that of wild-type cells. In wild-type , ACS activity was induced upon growth on ethanol, 2,3-butanediol, malonate and acetate. The wild-type and mutants defective in ACS activity showed an active acetate kinase (ACK) under the growth conditions used; however, phosphotransacetylase (PTA) could not be detected. The data indicate that requires active gene product for growth on ethanol.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-10-2671
2001-10-01
2020-04-07
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/10/1472671a.html?itemId=/content/journal/micro/10.1099/00221287-147-10-2671&mimeType=html&fmt=ahah

References

  1. Aceti D. J., Ferry J. G.. 1988; Purification and characterization of acetate kinase from acetate-grown Methanosarcina thermophila . Evidence for regulation of synthesis. J Biol Chem263:15444–15448
    [Google Scholar]
  2. 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 Res25:3389–3402[CrossRef]
    [Google Scholar]
  3. Bairoch A., Bucher P., Hofmann K.. 1997; The prosite database, its status in 1997. Nucleic Acids Res25:217–221[CrossRef]
    [Google Scholar]
  4. van den Berg M. A., Steensma H. Y.. 1995; ACS2, a Saccharomyces cerevisiae gene encoding acetyl-coenzyme A synthetase, essential for growth on glucose. Eur J Biochem231:704–713[CrossRef]
    [Google Scholar]
  5. Berg P.. 1962; Assay and preparation of yeast aceto-CoA-kinase. Methods Enzymol5:461–467
    [Google Scholar]
  6. Boyer H. W., Roulland-Dussoix D.. 1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli . J Mol Biol14:459–472
    [Google Scholar]
  7. Brown T. D. K., Jones-Mortimer M. C., Kornberg H. L.. 1977; The enzymic interconversion of acetate and acetyl-coenzyme A in Escherichia coli . J Gen Microbiol102:327–336[CrossRef]
    [Google Scholar]
  8. Cetin E. T., Töreci K. I., Ang Ö.. 1965; Encapsulated Pseudomonas aeruginosa ( Pseudomonas mucosus ) strains. J Bacteriol89:1432–1433
    [Google Scholar]
  9. Champine J. E., Goodwin S.. 1991; Acetate catabolism in the dissimilatory iron-reducing isolate GS-15. J Bacteriol173:2704–2706
    [Google Scholar]
  10. Chang D.-E., Shin S., Rhee J.-S., Pan J.-G.. 1999; Acetate metabolism in a pta mutant of Escherichia coli W3110: importance of maintaining acetyl coenzyme A flux for growth and survival. J Bacteriol181:6656–6663
    [Google Scholar]
  11. Connerton I. F., Fincham J. R. S., Sandeman R. A., Hynes M. J.. 1990; Comparison and cross-species expression of the acetyl-CoA synthetase genes of the ascomycete fungi, Aspergillus nidulans and Neurospora crassa . Mol Microbiol4:451–460[CrossRef]
    [Google Scholar]
  12. 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 Biochem257:409–419[CrossRef]
    [Google Scholar]
  13. Eaton R. W.. 1996; P-Cumate catabolic pathway in Pseudomonas putida F1: cloning and characterization of DNA carrying the cmt operon. J Bacteriol178:1351–1362
    [Google Scholar]
  14. Eggen R. I. L., Geerling A. C. M., Boshoven A. B. P., de Vos W. M.. 1991; Cloning, sequence analysis, and functional expression of the acetyl coenzyme A synthetase gene from Methanothrix soehngenii in Escherichia coli . J Bacteriol173:6383–6389
    [Google Scholar]
  15. 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 USA76:1648–1652[CrossRef]
    [Google Scholar]
  16. Groves W. E., Davis F. C., Sells B. H.. 1968; Spectrophotometric determination of microgram quantities of protein without nucleic acid interference. Anal Biochem22:195–210[CrossRef]
    [Google Scholar]
  17. Grundy F. J., Waters D. A., Takova T. Y., Henkin T. M.. 1993; Identification of genes involved in utilization of acetate and acetoin in Bacillus subtilis . Mol Microbiol10:259–271[CrossRef]
    [Google Scholar]
  18. Hanahan D.. 1983; Studies on transformation of Escherichia coli with plasmids. J Mol Biol166:557–580[CrossRef]
    [Google Scholar]
  19. Jetten M. S. M., Stams A. J. M., Zehnder A. J. B.. 1989; Isolation and characterization of acetyl-coenzyme A synthetase from Methanothrix soehngenii . J Bacteriol171:5430–5435
    [Google Scholar]
  20. Jones M. E., Lipmann F.. 1955; Aceto-CoA-kinase. Methods Enzymol1:585–590
    [Google Scholar]
  21. Kumari S., Tishel R., Eisenbach M., Wolfe A. J.. 1995; Cloning, characterization, and functional expression of acs, the gene which encodes acetyl coenzyme A synthetase in Escherichia coli. J Bacteriol 177:2878–2886
    [Google Scholar]
  22. Oberlies G., Fuchs G., Thauer R. K.. 1980; Acetate thiokinase and the assimilation of acetate in Methanobacterium thermoautotrophicum . Arch Microbiol128:248–252[CrossRef]
    [Google Scholar]
  23. Powlowski J., Sahlman L., Shingler V.. 1993; Purification and properties of the physically associated meta-cleavage pathway enzymes 4-hydroxy-2-ketovalerate aldolase and aldehyde dehydrogenase (acylating) from Pseudomonas sp. strain CF600. J Bacteriol175:377–385
    [Google Scholar]
  24. Priefert H., Steinbüchel A.. 1992; Identification and molecular characterization of the acetyl coenzyme A synthetase gene ( acoE ) of Alcaligenes eutrophus . J Bacteriol174:6590–6599
    [Google Scholar]
  25. Reichmann P., Görisch H.. 1993; Cytochrome c550 from Pseudomonas aeruginosa . Biochem J289:173–178
    [Google Scholar]
  26. Rupp M., Görisch H.. 1988; Purification, crystallization and characterization of quinoprotein ethanol dehydrogenase from Pseudomonas aeruginosa . Biol Chem Hoppe-Seyler369:431–439[CrossRef]
    [Google Scholar]
  27. 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]
  28. 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. Microbiology145:471–481[CrossRef]
    [Google Scholar]
  29. Schweizer H. P., Klassen T. R., Hoang T.. 1996; Improved methods for gene analysis in Pseudomonas . In Molecular Biology of Pseudomonads pp229–237 Edited by Nakazawa T., Furukawa K., Haas D., Silver S.. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  30. 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 Enzymol118:640–659
    [Google Scholar]
  31. Smith A. W., Iglewski B. H.. 1989; Transformation of Pseudomonas aeruginosa by electroporation. Nucleic Acids Res17:10509[CrossRef]
    [Google Scholar]
  32. 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 Bacteriol169:5789–5794
    [Google Scholar]
  33. Steinbüchel A., Fründ C., Jendrossek D., Schlegel H. G.. 1987; Isolation of mutants of Alcaligenes eutrophus unable to derepress the fermentative alcohol dehydrogenase. Arch Microbiol148:178–186[CrossRef]
    [Google Scholar]
  34. Taylor I. J., Anthony C.. 1976; Acetyl-CoA production and utilization during growth of the facultative methylotroph Pseudomonas AM1 on ethanol, malonate and 3-hydroxybutyrate. J Gen Microbiol95:134–143[CrossRef]
    [Google Scholar]
  35. Thompson D. K., Chen J. S.. 1990; Purification and properties of an acetoacetyl coenzyme A-reacting phosphotransbutyrylase from Clostridium beijerinckii . Appl Environ Microbiol56:607–613
    [Google Scholar]
  36. Wang S., Nakashima S., Numata O., Fujiu K., Nozawa Y.. 1999; Molecular cloning and cell-cycle-dependent expression of the acetyl-CoA synthetase gene in Tetrahymena cells. Biochem J343:479–485[CrossRef]
    [Google Scholar]
  37. Yanisch-Perron C., Vieira J., Messing J.. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13mp18 and pUC19 vectors. Gene33:103–119[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-10-2671
Loading
/content/journal/micro/10.1099/00221287-147-10-2671
Loading

Data & Media loading...

Most cited this month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error