1887

Abstract

UWD is a mutant of strain UW that is defective in the respiratory oxidation of NADH. This mutation causes an overproduction of polyhydroxyalkanoates (PHAs), as polyester synthesis is used as an alternative electron sink. Since PHAs have potential for use as natural, biodegradable plastics, studies of physiology related to their production are of interest. Alginate production by this strain is limited to <11 μg (mg cell protein), which permits high efficiency conversion of carbon source into PHA. However, ≤400 μg (mg cell protein) was formed when UWD cells were oxygen-limited and in the stationary phase of growth. Alginate formation was fuelled by PHA turnover, which was coincident with the synthesis of alkyl resorcinols, under conditions of exogenous glucose limitation. However, alginate production was a phenotypic and reversible change. Alginate production was stopped by interruption of with Tn. LacZ activity in UWD was shown to increase in stationary phase, while LacZ activity in a similarly constructed mutant of strain UW did not. Transcription of in strain UWD started from a previously identified RpoD promoter and not from the AlgU (RpoE) promoter. This is because strain UWD has a natural insertion element in . Differences between strain UW and UWD may reside in the defective respiratory oxidation of NADH, where the NADH surplus in strain UWD may act as a signal of stationary phase. Indeed, a backcross of UW DNA into UWD generated NADH-oxidase-proficient cells that failed to form alginate in stationary phase. Evidence is also presented to show that the RpoD promoter may be recognized by the stationary phase sigma factor (RpoS), which may mediate alginate production in strain UWD.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-2-483
2001-02-01
2020-01-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/2/1470483a.html?itemId=/content/journal/micro/10.1099/00221287-147-2-483&mimeType=html&fmt=ahah

References

  1. Anderson A., Dawes E.. 1990; Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev54:450–472
    [Google Scholar]
  2. Bauer C. E., Elsen S., Bird T. H.. 1999; Mechanisms for redox control of gene expression. Annu Rev Microbiol53:495–523[CrossRef]
    [Google Scholar]
  3. Beale J. M. Jr, Foster J. L.. 1996; Carbohydrate fluxes into alginate biosynthesis in Azotobacter vinelandii NCIB 8789: NMR investigations of the triose pools. Biochemistry35:4492–4501[CrossRef]
    [Google Scholar]
  4. Bergersen F.. 1980; Measurement of nitrogen fixation by direct means. In Methods for Evaluating Biological Nitrogen Fixation pp.65–110Edited by Bergersen F.. New York: Wiley;
    [Google Scholar]
  5. Bush J., Wilson P.. 1959; A non-gummy chromogenic strain of Azotobacter vinelandii. Nature184:381
    [Google Scholar]
  6. Campos M., Martinez-Salazar J. M., Lloret L., Moreno S., Nunez C., Espin G., Soberon-Chavez G.. 1996; Characterization of the gene coding for GDP-mannose dehydrogenase (algD) from Azotobacter vinelandii. J Bacteriol178:1793–1799
    [Google Scholar]
  7. Castaneda M., Guzman J., Moreno S., Espin G.. 2000; The GacS sensor kinase regulates alginate and poly-β-hydroxybutyrate production in Azotobacter vinelandii. J Bacteriol182:2624–2628[CrossRef]
    [Google Scholar]
  8. Chen G.-Q., Page W. J.. 1997; Production of poly-β-hydroxybutyrate by Azotobacter vinelandii in a two-stage fermentation process. Biotechnol Tech11:347–350[CrossRef]
    [Google Scholar]
  9. Collins S.. 1987; Choice of substrate in polyhydroxybutyrate synthesis. In Carbon Substrates in BiotechnologySpecial Publication of the Society for General Microbiology no. 21 pp.161–168Edited by Stowell J. D., Beardsmore A. J., Keevil C. W., Woodward J. R.. Oxford: IRL Press;
    [Google Scholar]
  10. Espinosa-Urgel M., Tormo A.. 1993; σs-dependent promoters in Escherichia coli are located in DNA regions with intrinsic curvature. Nucleic Acids Res21:3667–3670[CrossRef]
    [Google Scholar]
  11. Espinosa-Urgel M., Chamizo C., Tormo A.. 1996; A consensus structure for σS-dependent promoters. Mol Microbiol21:657–659[CrossRef]
    [Google Scholar]
  12. Fialho A. M., Zielinski N. A., Fett W. F., Chakrabarty A. M., Berry A.. 1990; Distribution of alginate gene sequences in the Pseudomonas rRNA homology group I-Azomonas-Azotobacter lineage of superfamily B procaryotes. Appl Environ Microbiol56:436–443
    [Google Scholar]
  13. Gacesa P.. 1998; Bacterial alginate biosynthesis – recent progress and future prospects. Microbiology144:1133–1143[CrossRef]
    [Google Scholar]
  14. Hengge-Aronis R.. 1996; Back to log phase: sigma S as a global regulator in the osmotic control of gene expression in Escherichia coli. Mol Microbiol21:887–893[CrossRef]
    [Google Scholar]
  15. Knutson C., Jeanes A.. 1968; A new modification of the carbazole analysis: application to heteropolysaccharides. Anal Biochem24:470–481[CrossRef]
    [Google Scholar]
  16. Kozubek A., Tyman J.. 1995; Cereal grain resorcinolic lipids: mono and dienoic homologues are present in rye grains. Chem Phys Lipids78:29–35[CrossRef]
    [Google Scholar]
  17. Law J., Slepecky R.. 1961; Assay of poly-β-hydroxybutyric acid. J Bacteriol82:33–36
    [Google Scholar]
  18. de Lorenzo V., Herrero M., Jakubzik U., Timmis K. N.. 1990; Mini-Tn5 transposon derivatives for insertional mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in Gram-negative eubacteria. J Bacteriol172:6568–6572
    [Google Scholar]
  19. Martinez-Salazar J. M., Moreno S., Najera R., Boucher J. C., Espin G., Soberon-Chavez G., Deretic V.. 1996; Characterization of the genes coding for the putative sigma factor AlgU and its regulators MucA, MucB, MucC, and MucD in Azotobacter vinelandii and evaluation of their roles in alginate biosynthesis. J Bacteriol178:1800–1808
    [Google Scholar]
  20. Miller J.. 1972; Experiments in Molecular Genetics Cold Spring Harbor NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  21. Neidhardt F., Ingraham J., Schaechter M.. 1990; Physiology of the Bacterial Cell. A Molecular Approach Sunderland MA: Sinauer Associates;
    [Google Scholar]
  22. Page W. J.. 1983; Formation of cystlike structures by iron-limited Azotobacter vinelandii strain UW during prolonged storage. Can J Microbiol29:1110–1118[CrossRef]
    [Google Scholar]
  23. Page W. J.. 1990; Production of poly-β-hydroxybutyrate by Azotobacter vinelandii UWD in beet molasses culture at high aeration. In Novel Biodegradable Microbial Polymers pp.423–424Edited by Dawes E.. Dordrecht: Kluwer Academic;
    [Google Scholar]
  24. Page W. J.. 1991; Examination of the role of Na+ in the physiology of the Na+-dependent soil bacterium Azotobacter salinestris. J Gen Microbiol137:2891–2899[CrossRef]
    [Google Scholar]
  25. Page W. J.. 1992; Production of polyhydroxyalkanoates by Azotobacter vinelandii UWD in beet molasses culture. FEMS Microbiol Rev103:149–158[CrossRef]
    [Google Scholar]
  26. Page W. J., Knosp O.. 1989; Hyperproduction of poly-β-hydroxybutyrate during exponential growth of Azotobacter vinelandii UWD. Appl Environ Microbiol55:1335–1339
    [Google Scholar]
  27. Page W. J., Manchak J.. 1994; Control of polyhydroxyalkanoate synthesis in Azotobacter vinelandii strain UWD. Microbiology140:953–963[CrossRef]
    [Google Scholar]
  28. Page W. J., Sadoff H. L.. 1976; Physiological factors affecting transformation of Azotobacter vinelandii. J Bacteriol125:1080–1087
    [Google Scholar]
  29. Page W. J., von Tigerstrom M.. 1979; Optimal conditions for transformation of Azotobacter vinelandii. J Bacteriol139:1058–1061
    [Google Scholar]
  30. Page W. J., Manchak J., Rudy B.. 1992; Formation of poly(hydroxybutyrate-co-hydroxyvalerate) by Azotobacter vinelandii UWD. Appl Environ Microbiol58:2866–2873
    [Google Scholar]
  31. Rehm B. H., Ertesvag H., Valla S.. 1996; A new Azotobacter vinelandii mannuronan C-5-epimerase gene (algG) is part of an alg gene cluster physically organized in a manner similar to that in Pseudomonas aeruginosa. J Bacteriol178:5884–5889
    [Google Scholar]
  32. Reusch R. N., Sadoff H. L.. 1981; Lipid metabolism during encystment of Azotobacter vinelandii. J Bacteriol145:889–895
    [Google Scholar]
  33. Robson R., Chesshyre J., Wheeler C., Jones R., Woodley P., Postgate J.. 1984; Genome size and complexity in Azotobacter chroococcum. J Gen Microbiol130:1603–1612
    [Google Scholar]
  34. Southern E.. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol98:503–517[CrossRef]
    [Google Scholar]
  35. Suh S. J., Silo-Suh L., Woods D. E., Hassett D. J., West S. E., Ohman D. E.. 1999; Effect of rpoS mutation on the stress response and expression of virulence factors in Pseudomonas aeruginosa. J Bacteriol181:3890–3897
    [Google Scholar]
  36. Tindale A. E., Mehrotra M., Ottem D., Page W. J.. 2000; Dual regulation of catecholate siderophore biosynthesis in Azotobacter vinelandii by iron and oxidative stress. Microbiology146:1617–1626
    [Google Scholar]
  37. Tluscik F., Kozubek A., Mejbaum-Katzenellenbogen W.. 1981; Alkylresorcinols in rye (secale cereale L.) grains. VI. Colorimetric micromethod for determination of alkylresorcinols with the use of diazonium salt, Fast Blue B. Acta Soc Bot Polon50:645–651
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-2-483
Loading
/content/journal/micro/10.1099/00221287-147-2-483
Loading

Data & Media loading...

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