This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Moiseeva, O. V.
Right arrow Articles by Schlömann, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Moiseeva, O. V.
Right arrow Articles by Schlömann, M.

 Previous Article  |  Next Article 

Journal of Bacteriology, October 2002, p. 5282-5292, Vol. 184, No. 19
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.19.5282-5292.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

A New Modified ortho Cleavage Pathway of 3-Chlorocatechol Degradation by Rhodococcus opacus 1CP: Genetic and Biochemical Evidence

Olga V. Moiseeva,1,2 Inna P. Solyanikova,1,2 Stefan R. Kaschabek,1,{dagger} Janosch Gröning,1,{dagger} Monika Thiel,1,{dagger} Ludmila A. Golovleva,2 and Michael Schlömann1*

Institut für Mikrobiologie, University of Stuttgart, 70550 Stuttgart, Germany,1 Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Science, Pushchino, Russia2

Received 25 March 2002/ Accepted 27 June 2002

The 4-chloro- and 2,4-dichlorophenol-degrading strain Rhodococcus opacus 1CP has previously been shown to acquire, during prolonged adaptation, the ability to mineralize 2-chlorophenol. In addition, homogeneous chlorocatechol 1,2-dioxygenase from 2-chlorophenol-grown biomass has shown relatively high activity towards 3-chlorocatechol. Based on sequences of the N terminus and tryptic peptides of this enzyme, degenerate PCR primers were now designed and used for cloning of the respective gene from genomic DNA of strain 1CP. A 9.5-kb fragment containing nine open reading frames was obtained on pROP1. Besides other genes, a gene cluster consisting of four chlorocatechol catabolic genes was identified. As judged by sequence similarity and correspondence of predicted N termini with those of purified enzymes, the open reading frames correspond to genes for a second chlorocatechol 1,2-dioxygenase (ClcA2), a second chloromuconate cycloisomerase (ClcB2), a second dienelactone hydrolase (ClcD2), and a muconolactone isomerase-related enzyme (ClcF). All enzymes of this new cluster are only distantly related to the known chlorocatechol enzymes and appear to represent new evolutionary lines of these activities. UV overlay spectra as well as high-pressure liquid chromatography analyses confirmed that 2-chloro-cis,cis-muconate is transformed by ClcB2 to 5-chloromuconolactone, which during turnover by ClcF gives cis-dienelactone as the sole product. cis-Dienelactone was further hydrolyzed by ClcD2 to maleylacetate. ClcF, despite its sequence similarity to muconolactone isomerases, no longer showed muconolactone-isomerizing activity and thus represents an enzyme dedicated to its new function as a 5-chloromuconolactone dehalogenase. Thus, during 3-chlorocatechol degradation by R. opacus 1CP, dechlorination is catalyzed by a muconolactone isomerase-related enzyme rather than by a specialized chloromuconate cycloisomerase.

* Corresponding author. Present address: TU Bergakademie Freiberg, IÖZ, 09596 Freiberg, Germany. Phone: 49-3731-393739. Fax: 49-3731-393012. E-mail: schloemann{at}ioez.tu-freiberg.de.

{dagger} Present address: TU Bergakademie Freiberg, IÖZ, 09596 Freiberg, Germany.

Journal of Bacteriology, October 2002, p. 5282-5292, Vol. 184, No. 19
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.19.5282-5292.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Camara, B., Nikodem, P., Bielecki, P., Bobadilla, R., Junca, H., Pieper, D. H. (2009). Characterization of a Gene Cluster Involved in 4-Chlorocatechol Degradation by Pseudomonas reinekei MT1. J. Bacteriol. 191: 4905-4915 [Abstract] [Full Text]  
  • Tischler, D., Eulberg, D., Lakner, S., Kaschabek, S. R., van Berkel, W. J. H., Schlomann, M. (2009). Identification of a Novel Self-Sufficient Styrene Monooxygenase from Rhodococcus opacus 1CP. J. Bacteriol. 191: 4996-5009 [Abstract] [Full Text]  
  • Kotani, T., Yurimoto, H., Kato, N., Sakai, Y. (2007). Novel Acetone Metabolism in a Propane-Utilizing Bacterium, Gordonia sp. Strain TY-5. J. Bacteriol. 189: 886-893 [Abstract] [Full Text]  
  • Gullon, S., Olano, C., Abdelfattah, M. S., Brana, A. F., Rohr, J., Mendez, C., Salas, J. A. (2006). Isolation, characterization, and heterologous expression of the biosynthesis gene cluster for the antitumor anthracycline steffimycin.. Appl. Environ. Microbiol. 72: 4172-4183 [Abstract] [Full Text]  
  • Pollmann, K., Wray, V., Pieper, D. H. (2005). Chloromethylmuconolactones as Critical Metabolites in the Degradation of Chloromethylcatechols: Recalcitrance of 2-Chlorotoluene. J. Bacteriol. 187: 2332-2340 [Abstract] [Full Text]  
  • Konig, C., Eulberg, D., Groning, J., Lakner, S., Seibert, V., Kaschabek, S. R., Schlomann, M. (2004). A linear megaplasmid, p1CP, carrying the genes for chlorocatechol catabolism of Rhodococcus opacus 1CP. Microbiology 150: 3075-3087 [Abstract] [Full Text]  
  • Seibert, V., Thiel, M., Hinner, I.-S., Schlomann, M. (2004). Characterization of a gene cluster encoding the maleylacetate reductase from Ralstonia eutropha 335T, an enzyme recruited for growth with 4-fluorobenzoate. Microbiology 150: 463-472 [Abstract] [Full Text]  
  • Nikodem, P., Hecht, V., Schlomann, M., Pieper, D. H. (2003). New Bacterial Pathway for 4- and 5-Chlorosalicylate Degradation via 4-Chlorocatechol and Maleylacetate in Pseudomonas sp. Strain MT1. J. Bacteriol. 185: 6790-6800 [Abstract] [Full Text]  
  • Kirchner, U., Westphal, A. H., Muller, R., van Berkel, W. J. H. (2003). Phenol Hydroxylase from Bacillus thermoglucosidasius A7, a Two-protein Component Monooxygenase with a Dual Role for FAD. J. Biol. Chem. 278: 47545-47553 [Abstract] [Full Text]  
  • Solyanikova, I. P., Moiseeva, O. V., Boeren, S., Boersma, M. G., Kolomytseva, M. P., Vervoort, J., Rietjens, I. M. C. M., Golovleva, L. A., van Berkel, W. J. H. (2003). Conversion of 2-Fluoromuconate to cis-Dienelactone by Purified Enzymes of Rhodococcus opacus 1cp. Appl. Environ. Microbiol. 69: 5636-5642 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»