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 Smith, D. J.
Right arrow Articles by Mohn, W. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Smith, D. J.
Right arrow Articles by Mohn, W. W.

 Previous Article  |  Next Article 

Journal of Bacteriology, June 2004, p. 3631-3639, Vol. 186, No. 11
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.11.3631-3639.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

A Cytochrome P450 Involved in the Metabolism of Abietane Diterpenoids by Pseudomonas abietaniphila BKME-9{dagger}

Daryl J. Smith,1 Vincent J. J. Martin,2 and William W. Mohn1*

Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada,1 Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 947202

Received 11 December 2003/ Accepted 26 February 2004

Diterpenoids are naturally occurring plant compounds which have pharmaceutical properties. We have sequenced a 10.4-kbp extension of the dit cluster in Pseudomonas abietaniphila BKME-9, containing genes involved in abietane diterpenoid biodegradation. The ditQ gene was found to encode a cytochrome P450 monooxygenase, P450dit, and to be homologous to the tdtD gene of Pseudomonas diterpeniphila A19-6a. Knocking out ditQ had little effect on growth of BKME-9 on abietic acid but severely impaired growth on dehydroabietic acid (DhA) and palustric acid (PaA), increasing doubling times from 3.8 to 15 h on DhA and from 5.6 to 18.5 h on PaA. A xylE transcriptional fusion showed that transcription of ditQ was induced by a range of diterpenoids. Substrate binding assays of P450dit expressed in Escherichia coli revealed that DhA binds to the enzyme and yields a type I binding spectrum with a Kd of 0.4 µM. These results indicate that P450dit is involved in the metabolism of DhA and PaA and are consistent with its putative role in converting DhA to 7-hydroxy-DhA. Finally, an amino acid sequence identity of greater than 72% and conserved gene arrangement support the hypothesis that the dit gene cluster of P. abietaniphila BKME-9 and the tdt cluster of P. diterpeniphila A19-6a contain functional homologues.

* 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.

{dagger} Dedicated to the memory of our colleague, R. C. Wyndham.

Journal of Bacteriology, June 2004, p. 3631-3639, Vol. 186, No. 11
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.11.3631-3639.2004
Copyright © 2004, 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]  
  • Chain, P. S. G., Denef, V. J., Konstantinidis, K. T., Vergez, L. M., Agullo, L., Reyes, V. L., Hauser, L., Cordova, M., Gomez, L., Gonzalez, M., Land, M., Lao, V., Larimer, F., LiPuma, J. J., Mahenthiralingam, E., Malfatti, S. A., Marx, C. J., Parnell, J. J., Ramette, A., Richardson, P., Seeger, M., Smith, D., Spilker, T., Sul, W. J., Tsoi, T. V., Ulrich, L. E., Zhulin, I. B., Tiedje, J. M. (2006). Inaugural Article: Burkholderia xenovorans LB400 harbors a multi-replicon, 9.73-Mbp genome shaped for versatility. Proc. Natl. Acad. Sci. USA 103: 15280-15287 [Abstract] [Full Text]  
  • Denef, V. J., Klappenbach, J. A., Patrauchan, M. A., Florizone, C., Rodrigues, J. L. M., Tsoi, T. V., Verstraete, W., Eltis, L. D., Tiedje, J. M. (2006). Genetic and Genomic Insights into the Role of Benzoate-Catabolic Pathway Redundancy in Burkholderia xenovorans LB400. Appl. Environ. Microbiol. 72: 585-595 [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»