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 Martin, V. J. J.
Right arrow Articles by Mohn, W. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Martin, V. J. J.
Right arrow Articles by Mohn, W. W.

 Previous Article  |  Next Article 

Journal of Bacteriology, May 1999, p. 2675-2682, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

A Novel Aromatic-Ring-Hydroxylating Dioxygenase from the Diterpenoid-Degrading Bacterium Pseudomonas abietaniphila BKME-9

Vincent J. J. Martin and William W. Mohn*

Department of Microbiology and Immunology and Pulp and Paper Center, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada

Received 27 October 1998/Accepted 16 February 1999

Pseudomonas abietaniphila BKME-9 is able to degrade dehydroabietic acid (DhA) via ring hydroxylation by a novel dioxygenase. The ditA1, ditA2, and ditA3 genes, which encode the alpha  and beta  subunits of the oxygenase and the ferredoxin of the diterpenoid dioxygenase, respectively, were isolated and sequenced. The ferredoxin gene is 9.2 kb upstream of the oxygenase genes and 872 bp upstream of a putative meta ring cleavage dioxygenase gene, ditC. A Tn5 insertion in the alpha  subunit gene, ditA1, resulted in the accumulation by the mutant strain BKME-941 of the pathway intermediate, 7-oxoDhA. Disruption of the ferredoxin gene, ditA3, in wild-type BKME-9 by mutant-allele exchange resulted in a strain (BKME-91) with a phenotype identical to that of the mutant strain BKME-941. Sequence analysis of the putative ferredoxin indicated that it is likely to be a [4Fe-4S]- or [3Fe-4S]-type ferredoxin and not a [2Fe-2S]-type ferredoxin, as found in all previously described ring-hydroxylating dioxygenases. Expression in Escherichia coli of ditA1A2A3, encoding the diterpenoid dioxygenase without its putative reductase component, resulted in a functional enzyme. The diterpenoid dioxygenase attacks 7-oxoDhA, and not DhA, at C-11 and C-12, producing 7-oxo-11,12-dihydroxy-8,13-abietadien acid, which was identified by 1H nuclear magnetic resonance, UV-visible light, and high-resolution mass spectrometry. The organization of the genes encoding the various components of the diterpenoid dioxygenase, the phylogenetic distinctiveness of both the alpha  subunit and the ferredoxin component, and the unusual Fe-S cluster of the ferredoxin all suggest that this enzyme belongs to a new class of aromatic ring-hydroxylating dioxygenases.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of British Columbia, 300-6174 University Blvd., Vancouver, B.C., Canada V6T 1Z3. Phone: (604) 822-4285. Fax: (604) 822-6041. E-mail: wmohn{at}interchange.ubc.ca.

Journal of Bacteriology, May 1999, p. 2675-2682, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Smith, D. J., Patrauchan, M. A., Florizone, C., Eltis, L. D., Mohn, W. W. (2008). Distinct Roles for Two CYP226 Family Cytochromes P450 in Abietane Diterpenoid Catabolism by Burkholderia xenovorans LB400. J. Bacteriol. 190: 1575-1583 [Abstract] [Full Text]  
  • Smith, D. J., Park, J., Tiedje, J. M., Mohn, W. W. (2007). A Large Gene Cluster in Burkholderia xenovorans Encoding Abietane Diterpenoid Catabolism. J. Bacteriol. 189: 6195-6204 [Abstract] [Full Text]  
  • Kim, S.-J., Kweon, O., Freeman, J. P., Jones, R. C., Adjei, M. D., Jhoo, J.-W., Edmondson, R. D., Cerniglia, C. E. (2006). Molecular Cloning and Expression of Genes Encoding a Novel Dioxygenase Involved in Low- and High-Molecular-Weight Polycyclic Aromatic Hydrocarbon Degradation in Mycobacterium vanbaalenii PYR-1. Appl. Environ. Microbiol. 72: 1045-1054 [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]  
  • Mukerjee-Dhar, G., Shimura, M., Miyazawa, D., Kimbara, K., Hatta, T. (2005). bph genes of the thermophilic PCB degrader, Bacillus sp. JF8: characterization of the divergent ring-hydroxylating dioxygenase and hydrolase genes upstream of the Mn-dependent BphC. Microbiology 151: 4139-4151 [Abstract] [Full Text]  
  • Kim, D., Chae, J.-C., Zylstra, G. J., Kim, Y.-S., Kim, S.-K., Nam, M. H., Kim, Y. M., Kim, E. (2004). Identification of a Novel Dioxygenase Involved in Metabolism of o-Xylene, Toluene, and Ethylbenzene by Rhodococcus sp. Strain DK17. Appl. Environ. Microbiol. 70: 7086-7092 [Abstract] [Full Text]  
  • Smith, D. J., Martin, V. J. J., Mohn, W. W. (2004). A Cytochrome P450 Involved in the Metabolism of Abietane Diterpenoids by Pseudomonas abietaniphila BKME-9. J. Bacteriol. 186: 3631-3639 [Abstract] [Full Text]  
  • Krivobok, S., Kuony, S., Meyer, C., Louwagie, M., Willison, J. C., Jouanneau, Y. (2003). Identification of Pyrene-Induced Proteins in Mycobacterium sp. Strain 6PY1: Evidence for Two Ring-Hydroxylating Dioxygenases. J. Bacteriol. 185: 3828-3841 [Abstract] [Full Text]  
  • Moreno-Ruiz, E., Hernaez, M. J., Martinez-Perez, O., Santero, E. (2003). Identification and Functional Characterization of Sphingomonas macrogolitabida Strain TFA Genes Involved in the First Two Steps of the Tetralin Catabolic Pathway. J. Bacteriol. 185: 2026-2030 [Abstract] [Full Text]  
  • Johnson, G. R., Jain, R. K., Spain, J. C. (2002). Origins of the 2,4-Dinitrotoluene Pathway. J. Bacteriol. 184: 4219-4232 [Abstract] [Full Text]  
  • Diaz, E., Ferrandez, A., Prieto, M. A., Garcia, J. L. (2001). Biodegradation of Aromatic Compounds by Escherichia coli. Microbiol. Mol. Biol. Rev. 65: 523-569 [Abstract] [Full Text]  
  • Haddad, S., Eby, D. M., Neidle, E. L. (2001). Cloning and Expression of the Benzoate Dioxygenase Genes from Rhodococcus sp. Strain 19070. Appl. Environ. Microbiol. 67: 2507-2514 [Abstract] [Full Text]  
  • Eby, D. M., Beharry, Z. M., Coulter, E. D., Kurtz, D. M. Jr., Neidle, E. L. (2001). Characterization and Evolution of Anthranilate 1,2-Dioxygenase from Acinetobacter sp. Strain ADP1. J. Bacteriol. 183: 109-118 [Abstract] [Full Text]  
  • Yu, Z., Stewart, G. R., Mohn, W. W. (2000). Apparent Contradiction: Psychrotolerant Bacteria from Hydrocarbon-Contaminated Arctic Tundra Soils That Degrade Diterpenoids Synthesized by Trees. Appl. Environ. Microbiol. 66: 5148-5154 [Abstract] [Full Text]  
  • Martin, V. J. J., Mohn, W. W. (2000). Genetic Investigation of the Catabolic Pathway for Degradation of Abietane Diterpenoids by Pseudomonas abietaniphila BKME-9. J. Bacteriol. 182: 3784-3793 [Abstract] [Full Text]  
  • Saito, A., Iwabuchi, T., Harayama, S. (2000). A Novel Phenanthrene Dioxygenase from Nocardioides sp. Strain KP7: Expression in Escherichia coli. J. Bacteriol. 182: 2134-2141 [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»