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 Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Happe, B Articles by Timmis, K N Search for Related Content PubMed PubMed Citation Articles by Happe, B Articles by Timmis, K N

research-article

Characterization of 2,2',3-trihydroxybiphenyl dioxygenase, an extradiol dioxygenase from the dibenzofuran- and dibenzo-p-dioxin-degrading bacterium Sphingomonas sp. strain RW1.

Department of Microbiology, GBF-National Research Center for Biotechnology, Braunschweig, Germany.

ABSTRACT

A key enzyme in the degradation pathways of dibenzo-p-dioxin and dibenzofuran, namely, 2,2',3-trihydroxybiphenyl dioxygenase, which is responsible for meta cleavage of the first aromatic ring, has been genetically and biochemically analyzed. The dbfB gene of this enzyme has been cloned from a cosmid library of the dibenzo-p-dioxin- and dibenzofuran-degrading bacterium Sphingomonas sp. strain RW1 (R. M. Wittich, H. Wilkes, V. Sinnwell, W. Francke, and P. Fortnagel, Appl. Environ. Microbiol. 58:1005-1010, 1992) and sequenced. The amino acid sequence of this enzyme is typical of those of extradiol dioxygenases. This enzyme, which is extremely oxygen labile, was purified anaerobically to apparent homogeneity from an Escherichia coli strain that had been engineered to hyperexpress dbfB. Unlike most extradiol dioxygenases, which have an oligomeric quaternary structure, the 2,2',3-trihydroxybiphenyl dioxygenase is a monomeric protein. Kinetic measurements with the purified enzyme produced similar Km values for 2,2',3-trihydroxybiphenyl and 2,3-dihydroxybiphenyl, and both of these compounds exhibited strong substrate inhibition. 2,2',3-Trihydroxydiphenyl ether, catechol, 3-methylcatechol, and 4-methylcatechol were oxidized less efficiently and 3,4-dihydroxybiphenyl was oxidized considerably less efficiently.

This article has been cited by other articles:

• 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]  
• Gai, Z., Yu, B., Li, L., Wang, Y., Ma, C., Feng, J., Deng, Z., Xu, P. (2007). Cometabolic Degradation of Dibenzofuran and Dibenzothiophene by a Newly Isolated Carbazole-Degrading Sphingomonas sp. Strain. Appl. Environ. Microbiol. 73: 2832-2838 [Abstract] [Full Text]  
• Fortin, P. D., Lo, A. T.-F., Haro, M.-A., Kaschabek, S. R., Reineke, W., Eltis, L. D. (2005). Evolutionarily Divergent Extradiol Dioxygenases Possess Higher Specificities for Polychlorinated Biphenyl Metabolites. J. Bacteriol. 187: 415-421 [Abstract] [Full Text]  
• Andujar, E., Santero, E. (2003). Site-directed mutagenesis of an extradiol dioxygenase involved in tetralin biodegradation identifies residues important for activity or substrate specificity. Microbiology 149: 1559-1567 [Abstract] [Full Text]  
• McKay, D. B., Prucha, M., Reineke, W., Timmis, K. N., Pieper, D. H. (2003). Substrate Specificity and Expression of Three 2,3-Dihydroxybiphenyl 1,2-Dioxygenases from Rhodococcus globerulus Strain P6. J. Bacteriol. 185: 2944-2951 [Abstract] [Full Text]  
• Gobel, M., Kassel-Cati, K., Schmidt, E., Reineke, W. (2002). Degradation of Aromatics and Chloroaromatics by Pseudomonas sp. Strain B13: Cloning, Characterization, and Analysis of Sequences Encoding 3-Oxoadipate:Succinyl-Coenzyme A (CoA) Transferase and 3-Oxoadipyl-CoA Thiolase. J. Bacteriol. 184: 216-223 [Abstract] [Full Text]  
• Riegert, U., Heiss, G., Kuhm, A. E., Müller, C., Contzen, M., Knackmuss, H.-J., Stolz, A. (1999). Catalytic Properties of the 3-Chlorocatechol-Oxidizing 2,3-Dihydroxybiphenyl 1,2-Dioxygenase from Sphingomonas sp. Strain BN6. J. Bacteriol. 181: 4812-4817 [Abstract] [Full Text]  
• Armengaud, J., Timmis, K. N., Wittich, R.-M. (1999). A Functional 4-Hydroxysalicylate/Hydroxyquinol Degradative Pathway Gene Cluster Is Linked to the Initial Dibenzo-p-Dioxin Pathway Genes in Sphingomonas sp. Strain RW1. J. Bacteriol. 181: 3452-3461 [Abstract] [Full Text]  
• Romine, M. F., Stillwell, L. C., Wong, K.-K., Thurston, S. J., Sisk, E. C., Sensen, C., Gaasterland, T., Fredrickson, J. K., Saffer, J. D. (1999). Complete Sequence of a 184-Kilobase Catabolic Plasmid from Sphingomonas aromaticivorans F199. J. Bacteriol. 181: 1585-1602 [Abstract] [Full Text]  
• Vaillancourt, F. H., Han, S., Fortin, P. D., Bolin, J. T., Eltis, L. D. (1998). Molecular Basis for the Stabilization and Inhibition of 2,3-Dihydroxybiphenyl 1,2-Dioxygenase by t-Butanol. J. Biol. Chem. 273: 34887-34895 [Abstract] [Full Text]  
• Armengaud, J., Happe, B., Timmis, K. N. (1998). Genetic Analysis of Dioxin Dioxygenase of Sphingomonas sp. Strain RW1: Catabolic Genes Dispersed on the Genome. J. Bacteriol. 180: 3954-3966 [Abstract] [Full Text]