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Applied and Environmental Microbiology, August 2002, p. 3716-3723, Vol. 68, No. 8
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.8.3716-3723.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Plasmid-Borne Genes Code for an Angular Dioxygenase Involved in Dibenzofuran Degradation by Terrabacter sp. Strain YK3

Toshiya Iida,1* Yuki Mukouzaka,1,2 Kaoru Nakamura,1 and Toshiaki Kudo1,3,4

Microbiology Laboratory, RIKEN (The Institute of Physical and Chemical Research),1 Bio-Recycle Project, Japan Science and Technology Corporation (JST), Wako, Saitama 351-0198,3 Graduate School of Science and Engineering, Saitama University, Saitama 338-0825,2 Science of Biological Supramolecular Systems, Graduate School of Integrated Science, Yokohama City University, Suehiro, Tsurumi-ku, Yokohama 230-0045, Japan4

Received 23 January 2002/ Accepted 30 April 2002

The genes responsible for angular dioxygenation of dibenzofuran in actinomycetes were cloned by using a degenerate set of PCR primers designed by using conserved sequences of the dioxygenase alpha subunit genes. One sequence of alpha subunit genes was commonly amplified from four dibenzofuran-utilizing actinomycetes: Terrabacter sp. strains YK1 and YK3, Rhodococcus sp. strain YK2, and Microbacterium sp. strain YK18. A 5.2-kb PstI fragment encoding the alpha and beta subunits of the terminal dioxygenase, ferredoxin, and ferredoxin reductase (designated dfdA1 to dfdA4, respectively) was cloned from the large circular plasmid pYK3 isolated from Terrabacter sp. strain YK3. We confirmed that transcription of the dfdA1 gene was induced by dibenzofuran in Terrabacter sp. strain YK3. Southern blot hybridization analysis revealed that this type of dioxygenase gene is distributed among diverse dibenzofuran-utilizing actinomycetes. However, genes homologous to dfdA1 were not detected in dibenzofuran utilization-deficient mutants of Terrabacter, Rhodococcus, and Microbacterium species. When the dfdA1 to dfdA4 genes were introduced into a non-dibenzofuran-degrading mutant of Rhodococcus sp. strain YK2, strain YK2-RD2, which had spontaneously lost the gene homologous to dfdA1, the ability to degrade dibenzofuran was restored. Analysis of the breakdown products indicated that DfdA has angular dioxygenase activity. This dfdA transformant degraded several aromatic compounds, indicating that the novel angular dioxygenase possesses broad substrate specificity.

* Corresponding author. Mailing address: Microbiology Laboratory, RIKEN (The Institute of Physical and Chemical Research), Wako, Saitama 351-0198, Japan. Phone: 81-(48)-462-1111, ext. 5725. Fax: 81-(48)-462-4672. E-mail: tiida{at}postman.riken.go.jp.

Applied and Environmental Microbiology, August 2002, p. 3716-3723, Vol. 68, No. 8
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.8.3716-3723.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Iida, T., Waki, T., Nakamura, K., Mukouzaka, Y., Kudo, T. (2009). The GAF-Like-Domain-Containing Transcriptional Regulator DfdR Is a Sensor Protein for Dibenzofuran and Several Hydrophobic Aromatic Compounds. J. Bacteriol. 191: 123-134 [Abstract] [Full Text]  
  • Aly, H. A. H., Huu, N. B., Wray, V., Junca, H., Pieper, D. H. (2008). Two Angular Dioxygenases Contribute to the Metabolic Versatility of Dibenzofuran-Degrading Rhodococcus sp. Strain HA01. Appl. Environ. Microbiol. 74: 3812-3822 [Abstract] [Full Text]  
  • Schuler, L., Ni Chadhain, S. M., Jouanneau, Y., Meyer, C., Zylstra, G. J., Hols, P., Agathos, S. N. (2008). Characterization of a Novel Angular Dioxygenase from Fluorene-Degrading Sphingomonas sp. Strain LB126. Appl. Environ. Microbiol. 74: 1050-1057 [Abstract] [Full Text]  


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