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 Fukumori, F Articles by Hausinger, R P Search for Related Content PubMed PubMed Citation Articles by Fukumori, F Articles by Hausinger, R P

research-article

Alcaligenes eutrophus JMP134 "2,4-dichlorophenoxyacetate monooxygenase" is an alpha-ketoglutarate-dependent dioxygenase.

Center for Microbial Ecology, Michigan State University, East Lansing 48824.

ABSTRACT

The Alcaligenes eutrophus JMP134 tfdA gene, encoding the enzyme responsible for the first step in 2,4-dichlorophenoxyacetic acid (2,4-D) biodegradation, was overexpressed in Escherichia coli, and several enzymatic properties of the partially purified gene product were examined. Although the tfdA-encoded enzyme is typically referred to as 2,4-D monooxygenase, we were unable to observe any reductant-dependent activity. Rather, we demonstrate that this enzyme is a ferrous ion-dependent dioxygenase that uses alpha-ketoglutarate as a cosubstrate. The alpha-ketoglutarate is converted to succinate concomitant with 2,4-D conversion to 2,4-dichlorophenol. By using [1-14C]alpha-ketoglutarate, we established that carbon dioxide is the second product derived from alpha-ketoglutarate. Finally, we verified the proposal that glyoxylate is the second product derived from 2,4-D.

This article has been cited by other articles:

• Larrain-Linton, J., De la Iglesia, R., Melo, F., Gonzalez, B. (2006). Molecular and Population Analyses of a Recombination Event in the Catabolic Plasmid pJP4.. J. Bacteriol. 188: 6793-6801 [Abstract] [Full Text]  
• Muller, T. A., Fleischmann, T., van der Meer, J. R., Kohler, H.-P. E. (2006). Purification and Characterization of Two Enantioselective {alpha}-Ketoglutarate-Dependent Dioxygenases, RdpA and SdpA, from Sphingomonas herbicidovorans MH.. Appl. Environ. Microbiol. 72: 4853-4861 [Abstract] [Full Text]  
• Muller, T. A., Zavodszky, M. I., Feig, M., Kuhn, L. A., Hausinger, R. P. (2006). Structural basis for the enantiospecificities of R- and S-specific phenoxypropionate/{alpha}-ketoglutarate dioxygenases.. Protein Sci. 15: 1356-1368 [Abstract] [Full Text]  
• Ledger, T., Pieper, D. H., Gonzalez, B. (2006). Chlorophenol Hydroxylases Encoded by Plasmid pJP4 Differentially Contribute to Chlorophenoxyacetic Acid Degradation. Appl. Environ. Microbiol. 72: 2783-2792 [Abstract] [Full Text]  
• Baelum, J., Henriksen, T., Hansen, H. C. B., Jacobsen, C. S. (2006). Degradation of 4-Chloro-2-Methylphenoxyacetic Acid in Top- and Subsoil Is Quantitatively Linked to the Class III tfdA Gene. Appl. Environ. Microbiol. 72: 1476-1486 [Abstract] [Full Text]  
• Muller, T. A., Byrde, S. M., Werlen, C., van der Meer, J. R., Kohler, H.-P. E. (2004). Genetic Analysis of Phenoxyalkanoic Acid Degradation in Sphingomonas herbicidovorans MH. Appl. Environ. Microbiol. 70: 6066-6075 [Abstract] [Full Text]  
• Shaw, L. J., Burns, R. G. (2004). Enhanced Mineralization of [U-14C]2,4-Dichlorophenoxyacetic Acid in Soil from the Rhizosphere of Trifolium pratense. Appl. Environ. Microbiol. 70: 4766-4774 [Abstract] [Full Text]  
• Itoh, K., Tashiro, Y., Uobe, K., Kamagata, Y., Suyama, K., Yamamoto, H. (2004). Root Nodule Bradyrhizobium spp. Harbor tfdA{alpha} and cadA, Homologous with Genes Encoding 2,4-Dichlorophenoxyacetic Acid-Degrading Proteins. Appl. Environ. Microbiol. 70: 2110-2118 [Abstract] [Full Text]  
• Hoffmann, D., Kleinsteuber, S., Muller, R. H., Babel, W. (2003). A transposon encoding the complete 2,4-dichlorophenoxyacetic acid degradation pathway in the alkalitolerant strain Delftia acidovorans P4a. Microbiology 149: 2545-2556 [Abstract] [Full Text]  
• White, A. K., Metcalf, W. W. (2002). Isolation and Biochemical Characterization of Hypophosphite/ 2-Oxoglutarate Dioxygenase. A NOVEL PHOSPHORUS-OXIDIZING ENZYME FROM PSEUDOMONAS STUTZERI WM88. J. Biol. Chem. 277: 38262-38271 [Abstract] [Full Text]  
• Plumeier, I., Perez-Pantoja, D., Heim, S., Gonzalez, B., Pieper, D. H. (2002). Importance of Different tfd Genes for Degradation of Chloroaromatics by Ralstonia eutropha JMP134. J. Bacteriol. 184: 4054-4064 [Abstract] [Full Text]  
• Itoh, K., Kanda, R., Sumita, Y., Kim, H., Kamagata, Y., Suyama, K., Yamamoto, H., Hausinger, R. P., Tiedje, J. M. (2002). tfdA-Like Genes in 2,4-Dichlorophenoxyacetic Acid-Degrading Bacteria Belonging to the Bradyrhizobium-Agromonas-Nitrobacter-Afipia Cluster in {alpha}-Proteobacteria. Appl. Environ. Microbiol. 68: 3449-3454 [Abstract] [Full Text]  
• Kitagawa, W., Takami, S., Miyauchi, K., Masai, E., Kamagata, Y., Tiedje, J. M., Fukuda, M. (2002). Novel 2,4-Dichlorophenoxyacetic Acid Degradation Genes from Oligotrophic Bradyrhizobium sp. Strain HW13 Isolated from a Pristine Environment. J. Bacteriol. 184: 509-518 [Abstract] [Full Text]  
• Clement, P., Pieper, D. H., Gonzalez, B. (2001). Molecular characterization of a deletion/duplication rearrangement in tfd genes from Ralstonia eutropha JMP134(pJP4) that improves growth on 3-chlorobenzoic acid but abolishes growth on 2,4-dichlorophenoxyacetic acid. Microbiology 147: 2141-2148 [Abstract] [Full Text]  
• Hay, A. G., Rice, J. F., Applegate, B. M., Bright, N. G., Sayler, G. S. (2000). A Bioluminescent Whole-Cell Reporter for Detection of 2,4-Dichlorophenoxyacetic Acid and 2,4-Dichlorophenol in Soil. Appl. Environ. Microbiol. 66: 4589-4594 [Abstract] [Full Text]  
• Laemmli, C. M., Leveau, J. H. J., Zehnder, A. J. B., van der Meer, J. R. (2000). Characterization of a Second tfd Gene Cluster for Chlorophenol and Chlorocatechol Metabolism on Plasmid pJP4 in Ralstonia eutropha JMP134(pJP4). J. Bacteriol. 182: 4165-4172 [Abstract] [Full Text]  
• Hogan, D. A., Smith, S. R., Saari, E. A., McCracken, J., Hausinger, R. P. (2000). Site-directed Mutagenesis of 2,4-Dichlorophenoxyacetic Acid/alpha -Ketoglutarate Dioxygenase. IDENTIFICATION OF RESIDUES INVOLVED IN METALLOCENTER FORMATION AND SUBSTRATE BINDING. J. Biol. Chem. 275: 12400-12409 [Abstract] [Full Text]  
• Pérez-Pantoja, D., Guzmán, L., Manzano, M., Pieper, D. H., González, B. (2000). Role of tfdCIDIEIFI and tfdDIICIIEIIFII Gene Modules in Catabolism of 3-Chlorobenzoate by Ralstonia eutropha JMP134(pJP4). Appl. Environ. Microbiol. 66: 1602-1608 [Abstract] [Full Text]  
• Müller, R. H., Babel, W. (2000). A Theoretical Study on the Metabolic Requirements Resulting from alpha -Ketoglutarate-Dependent Cleavage of Phenoxyalkanoates. Appl. Environ. Microbiol. 66: 339-344 [Abstract] [Full Text]  
• McGowan, C., Fulthorpe, R., Wright, A., Tiedje, J. M. (1998). Evidence for Interspecies Gene Transfer in the Evolution of 2,4-Dichlorophenoxyacetic Acid Degraders. Appl. Environ. Microbiol. 64: 4089-4092 [Abstract] [Full Text]  
• Leveau, J. H. J., Zehnder, A. J. B., van der Meer, J. R. (1998). The tfdK Gene Product Facilitates Uptake of 2,4-Dichlorophenoxyacetate by Ralstonia eutropha JMP134(pJP4). J. Bacteriol. 180: 2237-2243 [Abstract] [Full Text]  
• Eichhorn, E., van der Ploeg, J. R., Kertesz, M. A., Leisinger, T. (1997). Characterization of alpha -Ketoglutarate-dependent Taurine Dioxygenase from Escherichia coli. J. Biol. Chem. 272: 23031-23036 [Abstract] [Full Text]