Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 May;173(9):3010–3016. doi: 10.1128/jb.173.9.3010-3016.1991

Toluene-4-monooxygenase, a three-component enzyme system that catalyzes the oxidation of toluene to p-cresol in Pseudomonas mendocina KR1.

G M Whited 1, D T Gibson 1
PMCID: PMC207885  PMID: 2019563

Abstract

Pseudomonas mendocina KR1 grows on toluene as a sole carbon and energy source. A multicomponent oxygenase was partially purified from toluene-grown cells and separated into three protein components. The reconstituted enzyme system, in the presence of NADH and Fe2+, oxidized toluene to p-cresol as the first detectable product. Experiments with p-deutero-toluene led to the isolation of p-cresol which retained 68% of the deuterium initially present in the parent molecule. When the reconstituted enzyme system was incubated with toluene in the presence of 18O2, the oxygen in p-cresol was shown to be derived from molecular oxygen. The results demonstrate that P. mendocina KR1 initiates degradation of toluene by a multicomponent enzyme system which has been designated toluene-4-monooxygenase.

Full text

PDF
3010

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Axcell B. C., Geary P. J. Purification and some properties of a soluble benzene-oxidizing system from a strain of Pseudomonas. Biochem J. 1975 Jan;146(1):173–183. doi: 10.1042/bj1460173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Axcell B. C., Geary P. J. The metabolism of benzene by bacteria. Purification and some properties of the enzyme cis-1,2-dihydroxycyclohexa-3,5-diene (nicotinamide adenine dinucleotide) oxidoreductase (cis-benzene glycol dehydrogenase). Biochem J. 1973 Dec;136(4):927–934. doi: 10.1042/bj1360927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cerniglia C. E., Freeman J. P., Evans F. E. Evidence for an arene oxide-NIH shift pathway in the transformation of naphthalene to 1-naphthol by Bacillus cereus. Arch Microbiol. 1984 Aug;138(4):283–286. doi: 10.1007/BF00410891. [DOI] [PubMed] [Google Scholar]
  4. Colby J., Dalton H. Characterization of the second prosthetic group of the flavoenzyme NADH-acceptor reductase (component C) of the methane mono-oxygenase from Methylococcus capsulatus (Bath). Biochem J. 1979 Mar 1;177(3):903–908. doi: 10.1042/bj1770903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Colby J., Dalton H. Resolution of the methane mono-oxygenase of Methylococcus capsulatus (Bath) into three components. Purification and properties of component C, a flavoprotein. Biochem J. 1978 May 1;171(2):461–468. doi: 10.1042/bj1710461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Daly J. W., Jerina D. M., Witkop B. Arene oxides and the NIH shift: the metabolism, toxicity and carcinogenicity of aromatic compounds. Experientia. 1972 Oct 15;28(10):1129–1149. doi: 10.1007/BF01946135. [DOI] [PubMed] [Google Scholar]
  7. Ensley B. D., Gibson D. T., Laborde A. L. Oxidation of naphthalene by a multicomponent enzyme system from Pseudomonas sp. strain NCIB 9816. J Bacteriol. 1982 Mar;149(3):948–954. doi: 10.1128/jb.149.3.948-954.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fox B. G., Froland W. A., Dege J. E., Lipscomb J. D. Methane monooxygenase from Methylosinus trichosporium OB3b. Purification and properties of a three-component system with high specific activity from a type II methanotroph. J Biol Chem. 1989 Jun 15;264(17):10023–10033. [PubMed] [Google Scholar]
  9. Gibson D. T., Gschwendt B., Yeh W. K., Kobal V. M. Initial reactions in the oxidation of ethylbenzene by Pseudomonas putida. Biochemistry. 1973 Apr 10;12(8):1520–1528. doi: 10.1021/bi00732a008. [DOI] [PubMed] [Google Scholar]
  10. Gibson D. T., Roberts R. L., Wells M. C., Kobal V. M. Oxidation of biphenyl by a Beijerinckia species. Biochem Biophys Res Commun. 1973 Jan 23;50(2):211–219. doi: 10.1016/0006-291x(73)90828-0. [DOI] [PubMed] [Google Scholar]
  11. Guroff G., Reifsnyder C. A., Daly J. Retention of deuterium in p-tyrosine formed enzymatically from p-deuterophenylalanine. Biochem Biophys Res Commun. 1966 Sep 8;24(5):720–724. doi: 10.1016/0006-291x(66)90384-6. [DOI] [PubMed] [Google Scholar]
  12. Guroff G., Rhoads C. A. Phenylalanine hydroxylation by Pseudomonas species (ATCC 11299a). Nature of the cofactor. J Biol Chem. 1969 Jan 10;244(1):142–146. [PubMed] [Google Scholar]
  13. Guroff G., Strenkoski C. A. Biosynthesis of pteridines and of phenylalanine hydroxylase cofactor in cell-free extracts of Pseudomonas species (ATCC 11299a). J Biol Chem. 1966 May 25;241(10):2220–2227. [PubMed] [Google Scholar]
  14. Haigler B. E., Gibson D. T. Purification and properties of ferredoxinNAP, a component of naphthalene dioxygenase from Pseudomonas sp. strain NCIB 9816. J Bacteriol. 1990 Jan;172(1):465–468. doi: 10.1128/jb.172.1.465-468.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Heitkamp M. A., Freeman J. P., Miller D. W., Cerniglia C. E. Pyrene degradation by a Mycobacterium sp.: identification of ring oxidation and ring fission products. Appl Environ Microbiol. 1988 Oct;54(10):2556–2565. doi: 10.1128/aem.54.10.2556-2565.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jeffrey A. M., Yeh H. J., Jerina D. M., Patel T. R., Davey J. F., Gibson D. T. Initial reactions in the oxidation of naphthalene by Pseudomonas putida. Biochemistry. 1975 Feb 11;14(3):575–584. doi: 10.1021/bi00674a018. [DOI] [PubMed] [Google Scholar]
  17. Jerina D. M., Daly J. W., Witkop B. The role of arene oxide-oxepin systems in the metabolism of aromatic substrates. II. Synthesis of 3,4-toluene-4-2H oxide and subsequent "NIH shift" to 4-hydroxytoluene-3-2H. J Am Chem Soc. 1968 Nov 6;90(23):6523–6525. doi: 10.1021/ja01025a057. [DOI] [PubMed] [Google Scholar]
  18. Jerina D. M., Daly J. W., Witkop B., Zaltzman-Nirenberg P., Udenfriend S. The role of arene oxide-oxepin systems in the metabolism of aromatic substrates. 3. Formation of 1,2-naphthalene oxide from naphthalene by liver microsomes. J Am Chem Soc. 1968 Nov 6;90(23):6525–6527. doi: 10.1021/ja01025a058. [DOI] [PubMed] [Google Scholar]
  19. Kobal V. M., Gibson D. T., Davis R. E., Garza A. X-ray determination of the absolute stereochemistry of the initial oxidation product formed from toluene by Pseudomonas puida 39-D. J Am Chem Soc. 1973 Jun 27;95(13):4420–4421. doi: 10.1021/ja00794a048. [DOI] [PubMed] [Google Scholar]
  20. Kunz D. A., Chapman P. J. Catabolism of pseudocumene and 3-ethyltoluene by Pseudomonas putida (arvilla) mt-2: evidence for new functions of the TOL (pWWO) plasmid. J Bacteriol. 1981 Apr;146(1):179–191. doi: 10.1128/jb.146.1.179-191.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Lund J., Woodland M. P., Dalton H. Electron transfer reactions in the soluble methane monooxygenase of Methylococcus capsulatus (Bath). Eur J Biochem. 1985 Mar 1;147(2):297–305. doi: 10.1111/j.1432-1033.1985.tb08750.x. [DOI] [PubMed] [Google Scholar]
  23. Reddy C. C., Vaidyanathan C. S. Purification and properties of benzoate-4-hydroxylase from a soil pseudomonad. Arch Biochem Biophys. 1976 Dec;177(2):488–498. doi: 10.1016/0003-9861(76)90460-4. [DOI] [PubMed] [Google Scholar]
  24. Sauber K., Fröhner C., Rosenberg G., Eberspächer J., Lingens F. Purification and properties of pyrazon dioxygenase from pyrazon-degrading bacteria. Eur J Biochem. 1977 Mar 15;74(1):89–97. doi: 10.1111/j.1432-1033.1977.tb11370.x. [DOI] [PubMed] [Google Scholar]
  25. Shields M. S., Montgomery S. O., Chapman P. J., Cuskey S. M., Pritchard P. H. Novel pathway of toluene catabolism in the trichloroethylene-degrading bacterium g4. Appl Environ Microbiol. 1989 Jun;55(6):1624–1629. doi: 10.1128/aem.55.6.1624-1629.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stanier R. Y., Palleroni N. J., Doudoroff M. The aerobic pseudomonads: a taxonomic study. J Gen Microbiol. 1966 May;43(2):159–271. doi: 10.1099/00221287-43-2-159. [DOI] [PubMed] [Google Scholar]
  27. Subramanian V., Liu T. N., Yeh W. K., Serdar C. M., Wackett L. P., Gibson D. T. Purification and properties of ferredoxinTOL. A component of toluene dioxygenase from Pseudomonas putida F1. J Biol Chem. 1985 Feb 25;260(4):2355–2363. [PubMed] [Google Scholar]
  28. Whited G. M., Gibson D. T. Separation and partial characterization of the enzymes of the toluene-4-monooxygenase catabolic pathway in Pseudomonas mendocina KR1. J Bacteriol. 1991 May;173(9):3017–3020. doi: 10.1128/jb.173.9.3017-3020.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Worsey M. J., Williams P. A. Metabolism of toluene and xylenes by Pseudomonas (putida (arvilla) mt-2: evidence for a new function of the TOL plasmid. J Bacteriol. 1975 Oct;124(1):7–13. doi: 10.1128/jb.124.1.7-13.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yeh W. K., Gibson D. T., Liu T. N. Toluene dioxygenase: a multicomponent enzyme system. Biochem Biophys Res Commun. 1977 Sep 9;78(1):401–410. doi: 10.1016/0006-291x(77)91268-2. [DOI] [PubMed] [Google Scholar]
  31. Ziffer H., Jerina D. M., Gibson D. T., Kobal V. M. Absolute stereochemistry of the (+)-cis-1,2-dihydroxy-3-methylcyclohexa-3,5-diene produced from toluene by Pseudomonas putida. J Am Chem Soc. 1973 Jun 13;95(12):4048–4049. doi: 10.1021/ja00793a036. [DOI] [PubMed] [Google Scholar]
  32. Zürrer D., Cook A. M., Leisinger T. Microbial desulfonation of substituted naphthalenesulfonic acids and benzenesulfonic acids. Appl Environ Microbiol. 1987 Jul;53(7):1459–1463. doi: 10.1128/aem.53.7.1459-1463.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES