1887

Abstract

Three multicomponent oxygenases involved in the degradation of -toluenesulfonate and -toluenecarboxylate and the regulation of their synthesis have been examined in three strains (T-2, PSB-4 and TER-1) of Strain T-2 utilizes -toluenesulfonate as a source of carbon and energy for growth via -sulfobenzoate and protocatechuate, and -toluenecarboxylate via terephthalate and protocatechuate, and has the unusual property of requiring the reductase (TsaB) of the toluenesulfonate methyl monooxygenase system (TsaMB) in an incompletely expressed sulfobenzoate dioxygenase system (PsbAC) [ Schläfli Oppenberg, H. R., Chen, G., Leisinger, T. & Cook, A. M. (1995) . 141, 1891-1899]. The independently isolated PSB-4 utilized only sulfobenzoate and terephthalate via protocatechuate. Mutant TER-1, derived from strain T-2, utilized only terephthalate via protocatechuate. We detected no enzymes of the pathway from toluenesulfonate to sulfobenzoate in strains PSB-4 and TER-1, and confirmed by PCR and Southern blot analysis that the genes () encoding toluenesulfonate monooxygenase were absent. We concluded that, in strain PSB-4, the regulatory unit encoding the genes for the conversion of toluenesulfonate to sulfobenzoate was missing, and that generation of mutant TER-1 involved deletion of this regulatory unit and of the regulatory unit encoding desulfonation of sulfobenzoate. The degradation of sulfobenzoate in strain PSB-4 was catalysed by a fully inducible sulfobenzoate dioxygenase system (PsbAC), which, after purification of the oxygenase component (PsbA), turned out to be indistinguishable from the corresponding component from strain T-2 (PsbA). Reductase PsbC, which we could separate but not purify, was active with oxygenase PsbA and PsbA. Oxygenase PsbA was shown by electron paramagnetic resonance spectroscopy to contain a Rieske [2Fe-2S] centre. The enzyme system oxygenating terephthalate was examined and the oxygenase component purified and characterized. The oxygenase component in strains T-2 (and mutant TER-1) and PSB-4 were indistinguishable. The reductase component, which we separated but failed to purify, was active with the oxygenase from all strains. Gains and losses of blocks of genes in evolution is discussed.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-142-9-2419
1996-09-01
2021-05-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/9/mic-142-9-2419.html?itemId=/content/journal/micro/10.1099/00221287-142-9-2419&mimeType=html&fmt=ahah

References

  1. Assinder S. J., Williams P. A. 1990; The TOL plasmids: determinants of the catabolism of toluene and the xylenes. Adv Microb Physiol 31:1–69
    [Google Scholar]
  2. Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Seidman J. G., Smith J. A., Struhl K. 1987 Current Protocols in Molecular Biology. New York: Wiley;
    [Google Scholar]
  3. Batie C. J., Ballou D. P., Correll C. C. 1992; Phthalate dioxygenase reductase and related flavin-iron-sulfur containing electron transferases. In Chemistry and Biochemistry of Flavoenymes pp. 543–556 Müller F. Edited by Boca Raton: CRC Press;
    [Google Scholar]
  4. Bentley R. K., Holliman F. G. 1970; Pigments of Pseudomonas species. Part III. The synthesis of dimethylaeruginosin B and aeruginosin B. J Chem Soc(C) 1970:2447–2457
    [Google Scholar]
  5. Bloom H., Beyer H., Gross H. S. 1987; Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8:93–99
    [Google Scholar]
  6. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
    [Google Scholar]
  7. Busse H.-J., El-Banna T., Oyaizu H., Auling G. 1992; Identification of xenobiotic-degrading isolates from the beta subclass of the Proteobacteria by a polyphasic approach including 16S rRNA partial sequencing. Int J Syst Bacteriol 42:19–26
    [Google Scholar]
  8. Fewson C. A. 1981; Biodegradation of aromatics with industrial relevance. In Microbial Degradation of Xenobiotics and Recalcitrant Compounds pp. 141–179 Leisinger T., Cook A. M., Hütter R., Nüesch J. Edited by London: Academic Press;
    [Google Scholar]
  9. Furukawa K., Hirose L., Suyama A., Zaiki T., Hayashida S. 1993; Gene components responsible for discrete substrate specificity in the metabolism of biphenyl (bph operon) and toluene (tod operon). J Bacteriol 175:5224–5232
    [Google Scholar]
  10. Harayama S., Kok M., Neidle E. L. 1992; Functional and evolutionary relationships among diverse oxygenases. Annu Rev Microbiol 46:565–601
    [Google Scholar]
  11. Hirose J., Suyama A., Hayashida S., Furukawa K. 1994; Construction of hybrid biphenyl (bph) and toluene (tod) genes for functional analysis of aromatic ting dioxygenases. Gene 138:27–33
    [Google Scholar]
  12. Laemrnli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  13. Locher H. H., Leisinger T., Cook A. M. 1989; Degradation of p-toluenesulphonic acid via sidechain oxidation, desulphonation and meta ring cleavage in Pseudomonas (Comamonas) testosteroni T-2. J Gen Microbiol 135:1969–1978
    [Google Scholar]
  14. Locher H. H., Leisinger T., Cook A. M. 1991a; 4-Sulphobenzoate 3,4-dioxygenase: purification and properties of a desulphonative two-component enzyme system from Comamonas testosteroni T-2. Biochem J 274:833–842
    [Google Scholar]
  15. Locher H. H., Leisinger T., Cook A. M. 1991b; 4-Toluene sulfonate methyl-monooxygenase from Comamonas testosteroni T-2: purification and some properties of the oxygenase component. J Bacteriol 173:3741–3748
    [Google Scholar]
  16. Locher H. H., Malii C., Hooper S., Vorherr T., Leisinger T., Cook A. M. 1991c; Degradation of p-toluic acid (p-toluene carboxylic acid) and p-toluene sulphonic acid via oxygenation of the methyl sidechain is initiated by the same set of enzymes in C.omamonas testosteroni T-2. J Gen Microbiol 137:2201–2208
    [Google Scholar]
  17. van Loon W.M.G.M., Boon J. J., de Groot B. 1993; Quantitative analysis of sulfonic acid groups in macromolecular lignosulfonic acids and aquatic humic substances by temperature-resolved pyrolysis-mass spectrometry. Environ Sci Technol 27:2387–2396
    [Google Scholar]
  18. Mason J. R., Cammack R. 1992; The electron-transport proteins of hydroxylating bacterial dioxygenases. Anna Rev Microbiol 46:277–305
    [Google Scholar]
  19. Nakatsu C. H., Straus N. A., Wyndham R. C. 1995; Thenucleotide sequence of the Tn5271 3-chlorobenzoate 3,4-dioxygenase genes (cbaAB) unites the class IA oxygenases in a single lineage. Microbiology 141:485–495
    [Google Scholar]
  20. Riester J., Zumft W. G., Kroneck P. M. H. 1989; Nitrous oxide reductase from Pseudomonas stutzeri: redox properties and spectroscopic characterization of different forms of the multicopper enzyme. Eur J Biochem 178:751–762
    [Google Scholar]
  21. Saller E., Laue H., Schiäfli Oppenberg H. R., Cook A. M. 1995; Purification and some properties of (lR,2S)-l,2-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate dehydrogenase from Comamonas testosteroni T-2. FEMS Microbiol Lett 130:97–102
    [Google Scholar]
  22. Sehiäfli H. R., Weiss M. A., Leisinger T., Cook A. M. 1994; Terephthalate 1,2-dioxygenase system from Comamonas testosteroni T-2: purification and some properties of the oxygenase component. J Bacteriol 176:6644–6652
    [Google Scholar]
  23. Sehiäfli H. R., Baker D. P., Leisinger T., Cook A. M. 1995; Stereospecificity of hydride removal from NADH by reductases of multicomponent nonheme iron oxygenase systems. J Bacteriol 177:831–834
    [Google Scholar]
  24. Sehiäfli Oppenberg H. R., Chen G., Leisinger T., Cook A. M. 1995; Regu lation of the degradative pathways from 4-toluene-sulfonate and 4-toluenecarboxylate to protocatechuate in Comamonas testosteroni T-2. Microbiology 141:1891–1899
    [Google Scholar]
  25. Stanier R. Y., Ornston L. N. 1973; The β-ketoadipate pathway. Adv Microb Physiol 9:89–151
    [Google Scholar]
  26. Thurnheer T., Köthler T., Cook A. M., Leisinger T. 1986; Orthanilic acid and analogues as carbon sources for bacteria: growth physiology and enzymic desulphonation. . J Gen Microbiol 132:1215–1220
    [Google Scholar]
  27. Willems A., Pot B., Falsen E., Vandamme P., Gillis M., Kersters K., de Ley J. 1991; Polyphasic taxonomic study of the emended genus Comamonas: relationship to Aquaspiriilum aquattcum, E. Falsen group 10, and other clinical isolates. Int j Syst Bacteriol 41:427–444
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-142-9-2419
Loading
/content/journal/micro/10.1099/00221287-142-9-2419
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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