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. 1995 Jun;61(6):2308–2313. doi: 10.1128/aem.61.6.2308-2313.1995

Oxidative Pathway for the Biodegradation of Nitrobenzene by Comamonas sp. Strain JS765

S F Nishino, J C Spain
PMCID: PMC1388468  PMID: 16535050

Abstract

Previous studies have shown that the biodegradation of nitrobenzene by Pseudomonas pseudoalcaligenes JS45 proceeds by the reduction of nitrobenzene through nitrosobenzene and hydroxylaminobenzene, followed by rearrangement to 2-aminophenol, which then undergoes meta ring cleavage. We report here the isolation of a Comamonas sp. that uses an oxidative pathway for the complete mineralization of nitrobenzene. The isolate, designated strain JS765, uses nitrobenzene as a sole source of carbon, nitrogen, and energy. Nitrobenzene-grown cells oxidized nitrobenzene, with the stoichiometric release of nitrite. Extracts of nitrobenzene-grown JS765 showed high levels of catechol 2,3-dioxygenase activity that were not abolished by heating the cell extracts to 60(deg)C for 10 min. The ring cleavage product had an absorbance maximum at 375 nm, consistent with that of 2-hydroxymuconic semialdehyde. Both NAD-dependent dehydrogenase and NAD-independent hydrolase activities towards 2-hydroxymuconic semialdehyde were induced in extracts of nitrobenzene-grown cells. Catechol accumulated in the reaction mixture when cells preincubated with 3-chlorocatechol were incubated with nitrobenzene. Conversion of nitrobenzene to catechol by induced cells in the presence of 3-chlorocatechol and (sup18)O(inf2) demonstrated the simultaneous incorporation of two atoms of oxygen, which indicated that the initial reaction was dioxygenation. The results indicate that the catabolic pathway involves an initial dioxygenase attack on nitrobenzene with the release of nitrite and formation of catechol, which is subsequently degraded by a meta cleavage pathway.

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Selected References

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  1. An D., Gibson D. T., Spain J. C. Oxidative release of nitrite from 2-nitrotoluene by a three-component enzyme system from Pseudomonas sp. strain JS42. J Bacteriol. 1994 Dec;176(24):7462–7467. doi: 10.1128/jb.176.24.7462-7467.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bartels I., Knackmuss H. J., Reineke W. Suicide Inactivation of Catechol 2,3-Dioxygenase from Pseudomonas putida mt-2 by 3-Halocatechols. Appl Environ Microbiol. 1984 Mar;47(3):500–505. doi: 10.1128/aem.47.3.500-505.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bruhn C., Lenke H., Knackmuss H. J. Nitrosubstituted aromatic compounds as nitrogen source for bacteria. Appl Environ Microbiol. 1987 Jan;53(1):208–210. doi: 10.1128/aem.53.1.208-210.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. CARTWRIGHT N. J., CAIN R. B. Bacterial degradation of the nitrobenzoic acids. 2. Reduction of the nitro group. Biochem J. 1959 Oct;73:305–314. doi: 10.1042/bj0730305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. CARTWRIGHT N. J., CAIN R. B. Bacterial degradation of the nitrobenzoic acids. Biochem J. 1959 Feb;71(2):248–261. doi: 10.1042/bj0710248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Catterall F. A., Sala-Trepat J. M., Williams P. A. The coexistence of two pathways for the metabolism of 2-hydroxymuconic semialdehyde in a naphthalene-grown pseudomonad. Biochem Biophys Res Commun. 1971 May 7;43(3):463–469. doi: 10.1016/0006-291x(71)90636-x. [DOI] [PubMed] [Google Scholar]
  7. Dickel O., Knackmuss H. J. Catabolism of 1,3-dinitrobenzene by Rhodococcus sp. QT-1. Arch Microbiol. 1991;157(1):76–79. doi: 10.1007/BF00245339. [DOI] [PubMed] [Google Scholar]
  8. Groenewegen P. E., Breeuwer P., van Helvoort J. M., Langenhoff A. A., de Vries F. P., de Bont J. A. Novel degradative pathway of 4-nitrobenzoate in Comamonas acidovorans NBA-10. J Gen Microbiol. 1992 Aug;138(Pt 8):1599–1605. doi: 10.1099/00221287-138-8-1599. [DOI] [PubMed] [Google Scholar]
  9. Haigler B. E., Nishino S. F., Spain J. C. Biodegradation of 4-methyl-5-nitrocatechol by Pseudomonas sp. strain DNT. J Bacteriol. 1994 Jun;176(11):3433–3437. doi: 10.1128/jb.176.11.3433-3437.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Haigler B. E., Spain J. C. Biodegradation of 4-nitrotoluene by Pseudomonas sp. strain 4NT. Appl Environ Microbiol. 1993 Jul;59(7):2239–2243. doi: 10.1128/aem.59.7.2239-2243.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Haigler B. E., Wallace W. H., Spain J. C. Biodegradation of 2-nitrotoluene by Pseudomonas sp. strain JS42. Appl Environ Microbiol. 1994 Sep;60(9):3466–3469. doi: 10.1128/aem.60.9.3466-3469.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klecka G. M., Gibson D. T. Inhibition of catechol 2,3-dioxygenase from Pseudomonas putida by 3-chlorocatechol. Appl Environ Microbiol. 1981 May;41(5):1159–1165. doi: 10.1128/aem.41.5.1159-1165.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kukor J. J., Olsen R. H. Genetic organization and regulation of a meta cleavage pathway for catechols produced from catabolism of toluene, benzene, phenol, and cresols by Pseudomonas pickettii PKO1. J Bacteriol. 1991 Aug;173(15):4587–4594. doi: 10.1128/jb.173.15.4587-4594.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Murray K., Duggleby C. J., Sala-Trepat J. M., Williams P. A. The metabolism of benzoate and methylbenzoates via the meta-cleavage pathway by Pseudomonas arvilla mt-2. Eur J Biochem. 1972 Jul 24;28(3):301–310. doi: 10.1111/j.1432-1033.1972.tb01914.x. [DOI] [PubMed] [Google Scholar]
  15. Nadeau L. J., Spain J. C. Bacterial degradation of m-nitrobenzoic acid. Appl Environ Microbiol. 1995 Feb;61(2):840–843. doi: 10.1128/aem.61.2.840-843.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nishino S. F., Spain J. C., Belcher L. A., Litchfield C. D. Chlorobenzene degradation by bacteria isolated from contaminated groundwater. Appl Environ Microbiol. 1992 May;58(5):1719–1726. doi: 10.1128/aem.58.5.1719-1726.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nishino S. F., Spain J. C. Degradation of nitrobenzene by a Pseudomonas pseudoalcaligenes. Appl Environ Microbiol. 1993 Aug;59(8):2520–2525. doi: 10.1128/aem.59.8.2520-2525.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Peterson F. J., Mason R. P., Hovsepian J., Holtzman J. L. Oxygen-sensitive and -insensitive nitroreduction by Escherichia coli and rat hepatic microsomes. J Biol Chem. 1979 May 25;254(10):4009–4014. [PubMed] [Google Scholar]
  19. Rhys-Williams W., Taylor S. C., Williams P. A. A novel pathway for the catabolism of 4-nitrotoluene by Pseudomonas. J Gen Microbiol. 1993 Sep;139(9):1967–1972. doi: 10.1099/00221287-139-9-1967. [DOI] [PubMed] [Google Scholar]
  20. Sala-Trepat J. M., Evans W. C. The meta cleavage of catechol by Azotobacter species. 4-Oxalocrotonate pathway. Eur J Biochem. 1971 Jun 11;20(3):400–413. doi: 10.1111/j.1432-1033.1971.tb01406.x. [DOI] [PubMed] [Google Scholar]
  21. Schreiner A., Fuchs K., Lottspeich F., Poth H., Lingens F. Degradation of 2-methylaniline in Rhodococcus rhodochrous: cloning and expression of two clustered catechol 2,3-dioxygenase genes from strain CTM. J Gen Microbiol. 1991 Aug;137(8):2041–2048. doi: 10.1099/00221287-137-8-2041. [DOI] [PubMed] [Google Scholar]
  22. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  23. Spain J. C., Nishino S. F. Degradation of 1,4-dichlorobenzene by a Pseudomonas sp. Appl Environ Microbiol. 1987 May;53(5):1010–1019. doi: 10.1128/aem.53.5.1010-1019.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Spain J. C., Wyss O., Gibson D. T. Enzymatic oxidation of p-nitrophenol. Biochem Biophys Res Commun. 1979 May 28;88(2):634–641. doi: 10.1016/0006-291x(79)92095-3. [DOI] [PubMed] [Google Scholar]
  25. Spanggord R. J., Spain J. C., Nishino S. F., Mortelmans K. E. Biodegradation of 2,4-dinitrotoluene by a Pseudomonas sp. Appl Environ Microbiol. 1991 Nov;57(11):3200–3205. doi: 10.1128/aem.57.11.3200-3205.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zeyer J., Kocher H. P. Purification and characterization of a bacterial nitrophenol oxygenase which converts ortho-nitrophenol to catechol and nitrite. J Bacteriol. 1988 Apr;170(4):1789–1794. doi: 10.1128/jb.170.4.1789-1794.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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