This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bertoni, G. Articles by Barbieri, P. Search for Related Content PubMed PubMed Citation Articles by Bertoni, G. Articles by Barbieri, P. Agricola Articles by Bertoni, G. Articles by Barbieri, P.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., 10 1996, 3704-3711, Vol 62, No. 10
Copyright © 1996, American Society for Microbiology

Cloning of the genes for and characterization of the early stages of toluene and o-xylene catabolism in Pseudomonas stutzeri OX1

G Bertoni, F Bolognese, E Galli and P Barbieri
Dipartimento di Genetica e di Biologia dei Microrganismi, Universita degli Studi di Milano, Italy.

In order to study the toluene and o-xylene catabolic genes of Pseudomonas stutzeri OX1, a genomic library was constructed. A 28-kb EcoRI restriction endonuclease DNA fragment, cloned into the vector plasmid pLAFR1 and designated pFB3401, permitted Pseudomonas putida PaW340 to convert toluene and o-xylene into the corresponding meta-ring fission products. Physical and functional endonuclease restriction maps have been derived from the cloned DNA fragment. Further subcloning into and deletion analysis in the Escherichia coli vector pGEM-3Z allowed the genes for the conversion of toluene or o-xylene into the corresponding catechols to be mapped within a 6-kb region of the pFB3401 insert and their direction of transcription to be determined. Following exposure to toluene, E. coli cells carrying this 6-kb region produce a mixture of o-cresol, m-cresol, and p-cresol, which are further converted to 3-methylcatechol and 4-methylcatechol. Similarly, a mixture of 2,3-dimethylphenol and 3,4-dimethylphenol, further converted into dimethylcatechols, was detected after exposure to o- xylene. The enzyme involved in the first step of toluene and o-xylene degradation exhibited a broad substrate specificity, being able to oxidize also benzene, ethylbenzene, m-xylene, p-xylene, styrene, and naphthalene. Deletions of the 6-kb region which affect the ability to convert toluene or o-xylene into the corresponding methylphenols compromise also their further oxidation to methylcatechols. This suggests that a single enzyme system could be involved in both steps of the early stages of toluene and o-xylene catabolism.

This article has been cited by other articles:

• Lalucat, J., Bennasar, A., Bosch, R., Garcia-Valdes, E., Palleroni, N. J. (2006). Biology of Pseudomonas stutzeri. Microbiol. Mol. Biol. Rev. 70: 510-547 [Abstract] [Full Text]  
• Maruyama, T., Ishikura, M., Taki, H., Shindo, K., Kasai, H., Haga, M., Inomata, Y., Misawa, N. (2005). Isolation and Characterization of o-Xylene Oxygenase Genes from Rhodococcus opacus TKN14. Appl. Environ. Microbiol. 71: 7705-7715 [Abstract] [Full Text]  
• Cafaro, V., Notomista, E., Capasso, P., Di Donato, A. (2005). Regiospecificity of Two Multicomponent Monooxygenases from Pseudomonas stutzeri OX1: Molecular Basis for Catabolic Adaptation of This Microorganism to Methylated Aromatic Compounds. Appl. Environ. Microbiol. 71: 4736-4743 [Abstract] [Full Text]  
• Cafaro, V., Notomista, E., Capasso, P., Di Donato, A. (2005). Mutation of Glutamic Acid 103 of Toluene o-Xylene Monooxygenase as a Means To Control the Catabolic Efficiency of a Recombinant Upper Pathway for Degradation of Methylated Aromatic Compounds. Appl. Environ. Microbiol. 71: 4744-4750 [Abstract] [Full Text]  
• Vardar, G., Wood, T. K. (2005). Alpha-Subunit Positions Methionine 180 and Glutamate 214 of Pseudomonas stutzeri OX1 Toluene-o-Xylene Monooxygenase Influence Catalysis. J. Bacteriol. 187: 1511-1514 [Abstract] [Full Text]  
• Solera, D., Arenghi, F. L. G., Woelk, T., Galli, E., Barbieri, P. (2004). TouR-Mediated Effector-Independent Growth Phase-Dependent Activation of the {sigma}54 Ptou Promoter of Pseudomonas stutzeri OX1. J. Bacteriol. 186: 7353-7363 [Abstract] [Full Text]  
• Sazinsky, M. H., Bard, J., Di Donato, A., Lippard, S. J. (2004). Crystal Structure of the Toluene/o-Xylene Monooxygenase Hydroxylase from Pseudomonas stutzeri OX1: INSIGHT INTO THE SUBSTRATE SPECIFICITY, SUBSTRATE CHANNELING, AND ACTIVE SITE TUNING OF MULTICOMPONENT MONOOXYGENASES. J. Biol. Chem. 279: 30600-30610 [Abstract] [Full Text]  
• Tao, Y., Fishman, A., Bentley, W. E., Wood, T. K. (2004). Oxidation of Benzene to Phenol, Catechol, and 1,2,3-Trihydroxybenzene by Toluene 4-Monooxygenase of Pseudomonas mendocina KR1 and Toluene 3-Monooxygenase of Ralstonia pickettii PKO1. Appl. Environ. Microbiol. 70: 3814-3820 [Abstract] [Full Text]  
• Vardar, G., Wood, T. K. (2004). Protein Engineering of Toluene-o-Xylene Monooxygenase from Pseudomonas stutzeri OX1 for Synthesizing 4-Methylresorcinol, Methylhydroquinone, and Pyrogallol. Appl. Environ. Microbiol. 70: 3253-3262 [Abstract] [Full Text]  
• Roling, W. F. M., Milner, M. G., Jones, D. M., Fratepietro, F., Swannell, R. P. J., Daniel, F., Head, I. M. (2004). Bacterial Community Dynamics and Hydrocarbon Degradation during a Field-Scale Evaluation of Bioremediation on a Mudflat Beach Contaminated with Buried Oil. Appl. Environ. Microbiol. 70: 2603-2613 [Abstract] [Full Text]  
• Cafaro, V., Izzo, V., Scognamiglio, R., Notomista, E., Capasso, P., Casbarra, A., Pucci, P., Di Donato, A. (2004). Phenol Hydroxylase and Toluene/o-Xylene Monooxygenase from Pseudomonas stutzeri OX1: Interplay between Two Enzymes. Appl. Environ. Microbiol. 70: 2211-2219 [Abstract] [Full Text]  
• Jeong, J. J., Kim, J. H., Kim, C.-K., Hwang, I., Lee, K. (2003). 3- and 4-alkylphenol degradation pathway in Pseudomonas sp. strain KL28: genetic organization of the lap gene cluster and substrate specificities of phenol hydroxylase and catechol 2,3-dioxygenase. Microbiology 149: 3265-3277 [Abstract] [Full Text]  
• Kim, D., Kim, Y.-S., Kim, S.-K., Kim, S. W., Zylstra, G. J., Kim, Y. M., Kim, E. (2002). Monocyclic Aromatic Hydrocarbon Degradation by Rhodococcus sp. Strain DK17. Appl. Environ. Microbiol. 68: 3270-3278 [Abstract] [Full Text]  
• Arenghi, F. L. G., Berlanda, D., Galli, E., Sello, G., Barbieri, P. (2001). Organization and Regulation of meta Cleavage Pathway Genes for Toluene and o-Xylene Derivative Degradation in Pseudomonas stutzeri OX1. Appl. Environ. Microbiol. 67: 3304-3308 [Abstract] [Full Text]  
• Scognamiglio, R., Notomista, E., Barbieri, P., Pucci, P., Piaz, F. D., Tramontano, A., Di Donato, A. (2001). Conformational analysis of putative regulatory subunit D of the toluene/o-xylene-monooxygenase complex from Pseudomonas stutzeri OX1. Protein Sci. 10: 482-490 [Abstract] [Full Text]  
• Arenghi, F. L. G., Pinti, M., Galli, E., Barbieri, P. (1999). Identification of the Pseudomonas stutzeri OX1 Toluene-o-Xylene Monooxygenase Regulatory Gene (touR) and of Its Cognate Promoter. Appl. Environ. Microbiol. 65: 4057-4063 [Abstract] [Full Text]  
• Bolognese, F., di Lecce, C., Galli, E., Barbieri, P. (1999). Activation and Inactivation of Pseudomonas stutzeri Methylbenzene Catabolism Pathways Mediated by a Transposable Element. Appl. Environ. Microbiol. 65: 1876-1882 [Abstract] [Full Text]  
• Bertoni, G., Martino, M., Galli, E., Barbieri, P. (1998). Analysis of the Gene Cluster Encoding Toluene/o-Xylene Monooxygenase from Pseudomonas stutzeri OX1. Appl. Environ. Microbiol. 64: 3626-3632 [Abstract] [Full Text]  
• Chauhan, S., Barbieri, P., Wood, T. K. (1998). Oxidation of Trichloroethylene, 1,1-Dichloroethylene, and Chloroform by Toluene/o-Xylene Monooxygenase from Pseudomonas stutzeri OX1. Appl. Environ. Microbiol. 64: 3023-3024 [Abstract] [Full Text]