, Volume 21, Issue 6, pp 915–921 | Cite as

para-Nitrophenol 4-monooxygenase and hydroxyquinol 1,2-dioxygenase catalyze sequential transformation of 4-nitrocatechol in Pseudomonas sp. strain WBC-3

  • Min Wei
  • Jun-Jie Zhang
  • Hong Liu
  • Ning-Yi Zhou
Original Paper


Pseudomonas sp. strain WBC-3 utilizes para-nitrophenol (PNP) as a sole source of carbon, nitrogen and energy. PnpA (PNP 4-monooxygenase) and PnpB (para-benzoquinone reductase) were shown to be involved in the initial steps of PNP catabolism via hydroquinone. We demonstrated here that PnpA also catalyzed monooxygenation of 4-nitrocatechol (4-NC) to hydroxyquinol, probably via hydroxyquinone. It was the first time that a single-component PNP monooxygenase has been shown to catalyze this conversion. PnpG encoded by a gene located in the PNP degradation cluster was purified as a His-tagged protein and identified as a hydroxyquinol dioxygenase catalyzing a ring-cleavage reaction of hydroxyquinol. Although all the genes necessary for 4-NC metabolism seemed to be present in the PNP degradation cluster in strain WBC-3, it was unable to grow on 4-NC as a sole source of carbon, nitrogen and energy. This was apparently due to the substrate’s inability to trigger the expression of genes involved in degradation. Nevertheless, strain WBC-3 could completely degrade both PNP and 4-NC when PNP was used as the inducer, demonstrating its potential in bioremediation of the environment polluted by both 4-NC and PNP.


4-Nitrocatechol Hydroxyquinol 1,2-dioxygenase para-Nitrophenol 4-monooxygenase Pseudomonas Transformation 



We acknowledge financial supports from the National High Technology Research and Development Program of China (2006AA10Z403) and National Natural Science Foundation of China (30570021).


  1. Bohuslavek J, Chanama S, Crawford RL, Xun L (2005) Identification and characterization of hydroxyquinone hydratase activities from Sphingobium chlorophenolicum ATCC 39723. Biodegradation 16(4):353–362CrossRefPubMedGoogle Scholar
  2. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  3. Chapman PJ, Ribbons DW (1976) Metabolism of resorcinylic compounds by bacteria: alternative pathways for resorcinol catabolism in Pseudomonas putida. J Bacteriol 125(3):985–998PubMedGoogle Scholar
  4. Chauhan A, Samanta SK, Jain RK (2000) Degradation of 4-nitrocatechol by Burkholderia cepacia: a plasmid-encoded novel pathway. J Appl Microbiol 88(5):764–772CrossRefPubMedGoogle Scholar
  5. Duxbury JM, Tiedje JM, Alexander M, Dawson JE (1970) 2,4-D metabolism: enzymatic conversion of chloromaleylacetic acid to succinic acid. J Agric Food Chem 18(2):199–201CrossRefPubMedGoogle Scholar
  6. Johnson GR, Jain RK, Spain JC (2000) Properties of the trihydroxytoluene oxygenase from Burkholderia cepacia R34: an extradiol dioxygenase from the 2,4-dinitrotoluene pathway. Arch Microbiol 173(2):86–90CrossRefPubMedGoogle Scholar
  7. Kadiyala V, Spain JC (1998) A two-component monooxygenase catalyzes both the hydroxylation of p-nitrophenol and the oxidative release of nitrite from 4-nitrocatechol in Bacillus sphaericus JS905. Appl Environ Microbiol 64(7):2479–2484PubMedGoogle Scholar
  8. Kitagawa W, Kimura N, Kamagata Y (2004) A novel p-nitrophenol degradation gene cluster from a gram-positive bacterium, Rhodococcus opacus SAO101. J Bacteriol 186(15):4894–4902CrossRefPubMedGoogle Scholar
  9. Liu H, Zhang JJ, Wang SJ, Zhang XE, Zhou NY (2005) Plasmid-borne catabolism of methyl parathion and p-nitrophenol in Pseudomonas sp. strain WBC-3. Biochem Biophys Res Commun 334(4):1107–1114CrossRefPubMedGoogle Scholar
  10. Moonen MJ, Synowsky SA, van den Berg WA, Westphal AH, Heck AJ, van den Heuvel RH, Fraaije MW, van Berkel WJ (2008) Hydroquinone dioxygenase from Pseudomonas fluorescens ACB: a novel member of the family of nonheme-iron(II)-dependent dioxygenases. J Bacteriol 190(15):5199–5209CrossRefPubMedGoogle Scholar
  11. Patridge EV, Ferry JG (2006) WrbA from Escherichia coli and Archaeoglobus fulgidus is an NAD(P)H: quinone oxidoreductase. J Bacteriol 188(10):3498–3506CrossRefPubMedGoogle Scholar
  12. Spain JC, Gibson DT (1991) Pathway for biodegradation of p-nitrophenol in a Moraxella sp. Appl Environ Microbiol 57(3):812–819PubMedGoogle Scholar
  13. Spain JC, Wyss O, Gibson DT (1979) Enzymatic oxidation of p-nitrophenol. Biochem Biophys Res Commun 88(2):634–641CrossRefPubMedGoogle Scholar
  14. Sun L, Dong Y, Zhou Y, Yang M, Zhang C, Rao Z, Zhang XE (2004) Crystallization and preliminary X-ray studies of methyl parathion hydrolase from Pseudomonas sp. WBC-3. Acta Crystallogr D Biol Crystallogr 60(Pt 5):954–956CrossRefPubMedGoogle Scholar
  15. Takeo M, Murakami M, Niihara S, Yamamoto K, Nishimura M, Kato D, Negoro S (2008) Mechanism of 4-nitrophenol oxidation in Rhodococcus sp. Strain PN1: characterization of the two-component 4-nitrophenol hydroxylase and regulation of its expression. J Bacteriol 190(22):7367–7374CrossRefPubMedGoogle Scholar
  16. Zenno S, Koike H, Kumar AN, Jayaraman R, Tanokura M, Saigo K (1996) Biochemical characterization of NfsA, the Escherichia coli major nitroreductase exhibiting a high amino acid sequence homology to Frp, a Vibrio harveyi flavin oxidoreductase. J Bacteriol 178(15):4508–4514PubMedGoogle Scholar
  17. Zhang JJ, Liu H, Xiao Y, Zhang XE, Zhou NY (2009) Identification and characterization of catabolic para-nitrophenol 4-monooxygenase and para-benzoquinone reductase from Pseudomonas sp. strain WBC-3. J Bacteriol 191(8):2703–2710CrossRefPubMedGoogle Scholar
  18. Zhou NY, Fuenmayor SL, Williams PA (2001) nag genes of Ralstonia (formerly Pseudomonas) sp. strain U2 encoding enzymes for gentisate catabolism. J Bacteriol 183(2):700–708CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Min Wei
    • 1
  • Jun-Jie Zhang
    • 1
  • Hong Liu
    • 1
  • Ning-Yi Zhou
    • 1
  1. 1.State Key Laboratory of Virology, Wuhan Institute of VirologyChinese Academy of SciencesWuhanChina

Personalised recommendations