1887

Abstract

The transcriptional regulation of three distinct alcohol oxidation systems, alcohol dehydrogenase (ADH)-I, ADH-IIB and ADH-IIG, in HK5 was investigated under various induction conditions. The promoter activities of the genes involved in alcohol oxidation were determined using a transcriptional fusion promoter-probe vector. Ethanol was the best inducer for the divergent promoters of and , encoding ADH-I and a cytochrome , respectively. Primary and secondary C3 and C4 alcohols and butyraldehyde specifically induced the divergent promoters of and , encoding ADH-IIB and an NAD-dependent aldehyde dehydrogenase, respectively. The promoter of ADH-IIG responded well to ()-(+)-1,2-propanediol induction. In addition, the roles of genes encoding the response regulators and , located downstream of were inferred from the properties of - or -disrupted mutants and gene complementation tests. The gene products of both and were strictly necessary for transcription. The mutation and complementation studies also suggested a role for AgmR, but not ExaE, in the transcriptional regulation of (ADH-IIB) and (AGH-IIG). A hypothetical scheme describing a regulatory network, which directs expression of the three distinct alcohol oxidation systems in HK5, was derived.

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2009-02-01
2020-07-09
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References

  1. Anthony C.. 1982; The Biochemistry of Methylotrophs London: Academic Press;
  2. Anthony C., Williams P.. 2003; The structure and mechanism of methanol dehydrogenase. Biochim Biophys Acta1647:18–23
    [Google Scholar]
  3. Barrios H., Valderrama B., Morett E.. 1999; Compilation and analysis of σ 54-dependent promoter sequences. Nucleic Acids Res27:4305–4313
    [Google Scholar]
  4. Chen Z. W., Matsushita K., Yamashita T., Fujii T. A., Toyama H., Adachi O., Bellamy H. D., Mathews F. S.. 2002; Structure at 1.9 Å resolution of a quinohemoprotein alcohol dehydrogenase from Pseudomonas putida HK5. Structure10:837–849
    [Google Scholar]
  5. Choi K. H., Kumar A., Schweizer H. P.. 2006; A 10-min method for preparation of highly electrocompetent Pseudomonas aeruginosa cells: application for DNA fragment transfer between chromosomes and plasmid transformation. J Microbiol Methods64:391–397
    [Google Scholar]
  6. Chuang S. E., Daniels D. L., Blattner F. R.. 1993; Global regulation of gene expression in Escherichia coli . J Bacteriol175:2026–2036
    [Google Scholar]
  7. Dulley J. R., Grieve P. A.. 1975; A simple technique for eliminating interference by detergents in the Lowry method of protein determination. Anal Biochem64:136–141
    [Google Scholar]
  8. Farinha M. A., Kropinski A. M.. 1990; Construction of broad-host-range plasmid vectors for easy visible selection and analysis of promoters. J Bacteriol172:3496–3499
    [Google Scholar]
  9. Gliese N., Khodaverdi V., Schobert M., Görisch H.. 2004; AgmR controls transcription of a regulon with several operons essential for ethanol oxidation in Pseudomonas aeruginosa ATCC 17933. Microbiology150:1851–1857
    [Google Scholar]
  10. Görisch H.. 2003; The ethanol oxidation system and its regulation in Pseudomonas aeruginosa . Biochim Biophys Acta1647:98–102
    [Google Scholar]
  11. Harley C. B., Reynolds R. P.. 1987; Analysis of E. coli promoter sequences. Nucleic Acids Res15:2343–2361
    [Google Scholar]
  12. Lidstrom M. E., Anthony C., Biville F., Gasser F., Goodwin P., Hanson R. S., Harms N.. 1994; New unified nomenclature for genes involved in the oxidation of methanol in Gram-negative bacteria. FEMS Microbiol Lett117:103–106
    [Google Scholar]
  13. Marx C. J., Lidstrom M. E.. 2001; Development of improved versatile broad-host-range vectors for use in methylotrophs and other Gram-negative bacteria. Microbiology147:2065–2075
    [Google Scholar]
  14. Matsushita K., Yamashita T., Aoki N., Toyama H., Adachi O.. 1999; Electron transfer from quinohemoprotein alcohol dehydrogenase to blue copper protein azurin in the alcohol oxidase respiratory chain of Pseudomonas putida HK5. Biochemistry38:6111–6118
    [Google Scholar]
  15. Miller J. M.. 1992; A Short Course in Bacterial Genetics, a Laboratory Manual and Handbook for Escherichia coli and Related Bacteria Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  16. Promden W., Vangnai A. S., Pongsawasdi P., Adachi O., Matsushita K., Toyama H.. 2008; Disruption of quinoprotein ethanol dehydrogenase gene and adjacent genes in Pseudomonas putida HK5. FEMS Microbiol Lett280:203–209
    [Google Scholar]
  17. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual , 2nd eds. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  18. 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
    [Google Scholar]
  19. 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 . Microbiology147:363–372
    [Google Scholar]
  20. Springer A. L., Chou H. H., Fan W. H., Lee E., Lidstrom M. E.. 1995; Methanol oxidation mutants in Methylobacterium extorquens AM1: identification of new genetic complementation groups. Microbiology141:2985–2993
    [Google Scholar]
  21. Toyama H., Fujii A., Matsushita K., Shinagawa E., Ameyama M., Adachi O.. 1995; Three distinct quinoprotein alcohol dehydrogenases are expressed when Pseudomonas putida is grown on different alcohols. J Bacteriol177:2442–2450
    [Google Scholar]
  22. Toyama H., Fujii T., Aoki N., Matsushita K., Adachi O.. 2003; Molecular cloning of quinohemoprotein alcohol dehydrogenase, ADH IIB, from Pseudomonas putida HK5. Biosci Biotechnol Biochem67:1397–1400
    [Google Scholar]
  23. Toyama H., Mathews F. S., Adachi O., Matsushita K.. 2004; Quinohemoprotein alcohol dehydrogenases: structure, function, and physiology. Arch Biochem Biophys428:10–21
    [Google Scholar]
  24. Toyama H., Chen Z. W., Fukumoto M., Adachi O., Matsushita K., Mathews F. S.. 2005; Molecular cloning and structural analysis of quinohemoprotein alcohol dehydrogenase ADH-IIG from Pseudomonas putida HK5. J Mol Biol352:91–104
    [Google Scholar]
  25. Vangnai A. S., Arp D. J., Sayavedra-Soto L. A.. 2002; Two distinct alcohol dehydrogenases participate in butane metabolism by Pseudomonas butanovora . J Bacteriol184:1916–1924
    [Google Scholar]
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