Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Jun;176(11):3433–3437. doi: 10.1128/jb.176.11.3433-3437.1994

Biodegradation of 4-methyl-5-nitrocatechol by Pseudomonas sp. strain DNT.

B E Haigler 1, S F Nishino 1, J C Spain 1
PMCID: PMC205522  PMID: 8195105

Abstract

Pseudomonas sp. strain DNT degrades 2,4-dinitrotoluene (DNT) by a dioxygenase attack at the 4,5 position with concomitant removal of the nitro group to yield 4-methyl-5-nitrocatechol (MNC). Here we describe the mechanism of removal of the nitro group from MNC and subsequent reactions leading to ring fission. Washed suspensions of DNT-grown cells oxidized MNC and 2,4,5-trihydroxytoluene (THT). Extracts prepared from DNT-induced cells catalyzed the disappearance of MNC in the presence of oxygen and NADPH. Partially purified MNC oxygenase oxidized MNC in a reaction requiring 1 mol of NADPH and 1 mol of oxygen per mol of substrate. The enzyme converted MNC to 2-hydroxy-5-methylquinone (HMQ), which was identified by gas chromatography-mass spectrometry. HMQ was also detected transiently in culture fluids of cells grown on DNT. A quinone reductase was partially purified and shown to convert HMQ to THT in a reaction requiring NADH. A partially purified THT oxygenase catalyzed ring fission of THT and accumulation of a compound tentatively identified as 3-hydroxy-5-(1-formylethylidene)-2-furanone. Preliminary results indicate that this compound is an artifact of the isolation procedure and suggest that 2,4-dihydroxy-5-methyl-6-oxo-2,4-hexadienoic acid is the actual ring fission product.

Full text

PDF
3433

Selected References

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

  1. Bayly R. C., Dagley S., Gibson D. T. The metabolism of cresols by species of Pseudomonas. Biochem J. 1966 Nov;101(2):293–301. doi: 10.1042/bj1010293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cain R. B., Farr D. R. Metabolism of arylsulphonates by micro-organisms. Biochem J. 1968 Feb;106(4):859–877. doi: 10.1042/bj1060859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chapman P. J., Ribbons D. W. Metabolism of resorcinylic compounds by bacteria: alternative pathways for resorcinol catabolism in Pseudomonas putida. J Bacteriol. 1976 Mar;125(3):985–998. doi: 10.1128/jb.125.3.985-998.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chapman P. J., Ribbons D. W. Metabolism of resorcinylic compounds by bacteria: orcinol pathway in Pseudomonas putida. J Bacteriol. 1976 Mar;125(3):975–984. doi: 10.1128/jb.125.3.975-984.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Duque E., Haidour A., Godoy F., Ramos J. L. Construction of a Pseudomonas hybrid strain that mineralizes 2,4,6-trinitrotoluene. J Bacteriol. 1993 Apr;175(8):2278–2283. doi: 10.1128/jb.175.8.2278-2283.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Harayama S., Rekik M. Bacterial aromatic ring-cleavage enzymes are classified into two different gene families. J Biol Chem. 1989 Sep 15;264(26):15328–15333. [PubMed] [Google Scholar]
  8. Husain M., Entsch B., Ballou D. P., Massey V., Chapman P. J. Fluoride elimination from substrates in hydroxylation reactions catalyzed by p-hydroxybenzoate hydroxylase. J Biol Chem. 1980 May 10;255(9):4189–4197. [PubMed] [Google Scholar]
  9. Lenke H., Knackmuss H. J. Initial hydrogenation during catabolism of picric acid by Rhodococcus erythropolis HL 24-2. Appl Environ Microbiol. 1992 Sep;58(9):2933–2937. doi: 10.1128/aem.58.9.2933-2937.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Spain J. C., Gibson D. T. Pathway for Biodegradation of p-Nitrophenol in a Moraxella sp. Appl Environ Microbiol. 1991 Mar;57(3):812–819. doi: 10.1128/aem.57.3.812-819.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. 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]
  14. Suen W. C., Spain J. C. Cloning and characterization of Pseudomonas sp. strain DNT genes for 2,4-dinitrotoluene degradation. J Bacteriol. 1993 Mar;175(6):1831–1837. doi: 10.1128/jb.175.6.1831-1837.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Xun L., Topp E., Orser C. S. Diverse substrate range of a Flavobacterium pentachlorophenol hydroxylase and reaction stoichiometries. J Bacteriol. 1992 May;174(9):2898–2902. doi: 10.1128/jb.174.9.2898-2902.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]

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

RESOURCES