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 Fuenmayor, S. L.
Right arrow Articles by Williams, P. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fuenmayor, S. L.
Right arrow Articles by Williams, P. A.

 Previous Article  |  Next Article 

J Bacteriol, May 1998, p. 2522-2530, Vol. 180, No. 9
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

A Gene Cluster Encoding Steps in Conversion of Naphthalene to Gentisate in Pseudomonas sp. Strain U2

Sergio L. Fuenmayor,1 Mark Wild,2 Alastair L. Boyes,2 and Peter A. Williams1,*

School of Biological Sciences1 and Department of Chemistry,2 University of Wales, Bangor, Gwynedd LL57 2UW, Wales, United Kingdom

Received 14 October 1997/Accepted 23 February 1998

Pseudomonas sp. strain U2 was isolated from oil-contaminated soil in Venezuela by selective enrichment on naphthalene as the sole carbon source. The genes for naphthalene dioxygenase were cloned from the plasmid DNA of strain U2 on an 8.3-kb BamHI fragment. The genes for the naphthalene dioxygenase genes nagAa (for ferredoxin reductase), nagAb (for ferredoxin), and nagAc and nagAd (for the large and small subunits of dioxygenase, respectively) were located by Southern hybridizations and by nucleotide sequencing. The genes for nagB (for naphthalene cis-dihydrodiol dehydrogenase) and nagF (for salicylaldehyde dehydrogenase) were inferred from subclones by their biochemical activities. Between nagAa and nagAb were two open reading frames, homologs of which have also been identified in similar locations in two nitrotoluene-using strains (J. V. Parales, A. Kumar, R. E. Parales, and D. T. Gibson, Gene 181:57-61, 1996; W.-C. Suen, B. Haigler, and J. C. Spain, J. Bacteriol. 178:4926-4934, 1996) and a naphthalene-using strain (G. J. Zylstra, E. Kim, and A. K. Goyal, Genet. Eng. 19:257-269, 1997). Recombinant Escherichia coli strains with plasmids carrying this region were able to convert salicylate to gentisate, which was identified by a combination of gas chromatography-mass spectrometry and nuclear magnetic resonance. The first open reading frame, designated nagG, encodes a protein with characteristics of a Rieske-type iron-sulfur center homologous to the large subunits of dihydroxylating dioxygenases, and the second open reading frame, designated nagH, encodes a protein with limited homology to the small subunits of the same dioxygenases. Cloned together in E. coli, nagG, nagH, and nagAb, were able to convert salicylate (2-hydroxybenzoate) into gentisate (2,5-dihydroxybenzoate) and therefore encode a salicylate 5-hydroxylase activity. Single-gene knockouts of nagG, nagH, and nagAb demonstrated their functional roles in the formation of gentisate. It is proposed that NagG and NagH are structural subunits of salicylate 5-hydroxylase linked to an electron transport chain consisting of NagAb and NagAa, although E. coli appears to be able to partially substitute for the latter. This constitutes a novel mechanism for monohydroxylation of the aromatic ring. Salicylate hydroxylase and catechol 2,3-dioxygenase in strain U2 could not be detected either by enzyme assay or by Southern hybridization. However growth on both naphthalene and salicylate caused induction of gentisate 1,2-dioxygenase, confirming this route for salicylate catabolism in strain U2. Sequence comparisons suggest that the novel gene order nagAa-nagG-nagH-nagAb-nagAc-nagAd-nagB-nagF represents the archetype for naphthalene strains which use the gentisate pathway rather than the meta cleavage pathway of catechol.

* Corresponding author. Mailing address: School of Biological Sciences, Memorial Building, University of Wales, Bangor, Gwynedd LL57 2UW, Wales, United Kingdom. Phone: (44) 1248 382363. Fax: (44) 1248 370731. E-mail: P.A.Williams{at}bangor.ac.uk.

J Bacteriol, May 1998, p. 2522-2530, Vol. 180, No. 9
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Shin, K. A., Spain, J. C. (2009). Pathway and Evolutionary Implications of Diphenylamine Biodegradation by Burkholderia sp. Strain JS667. Appl. Environ. Microbiol. 75: 2694-2704 [Abstract] [Full Text]  
  • Gomes, N. C. M., Borges, L. R., Paranhos, R., Pinto, F. N., Krogerrecklenfort, E., Mendonca-Hagler, L. C. S., Smalla, K. (2007). Diversity of ndo Genes in Mangrove Sediments Exposed to Different Sources of Polycyclic Aromatic Hydrocarbon Pollution. Appl. Environ. Microbiol. 73: 7392-7399 [Abstract] [Full Text]  
  • Pumphrey, G. M., Madsen, E. L. (2007). Naphthalene metabolism and growth inhibition by naphthalene in Polaromonas naphthalenivorans strain CJ2. Microbiology 153: 3730-3738 [Abstract] [Full Text]  
  • Park, M., Jeon, Y., Jang, H. H., Ro, H.-S., Park, W., Madsen, E. L., Jeon, C. O. (2007). Molecular and Biochemical Characterization of 3-Hydroxybenzoate 6-Hydroxylase from Polaromonas naphthalenivorans CJ2. Appl. Environ. Microbiol. 73: 5146-5152 [Abstract] [Full Text]  
  • Poonthrigpun, S., Pattaragulwanit, K., Paengthai, S., Kriangkripipat, T., Juntongjin, K., Thaniyavarn, S., Petsom, A., Pinphanichakarn, P. (2006). Novel Intermediates of Acenaphthylene Degradation by Rhizobium sp. Strain CU-A1: Evidence for Naphthalene-1,8-Dicarboxylic Acid Metabolism. Appl. Environ. Microbiol. 72: 6034-6039 [Abstract] [Full Text]  
  • Feng, J., Che, Y., Milse, J., Yin, Y.-J., Liu, L., Ruckert, C., Shen, X.-H., Qi, S.-W., Kalinowski, J., Liu, S.-J. (2006). The Gene ncgl2918 Encodes a Novel Maleylpyruvate Isomerase That Needs Mycothiol as Cofactor and Links Mycothiol Biosynthesis and Gentisate Assimilation in Corynebacterium glutamicum. J. Biol. Chem. 281: 10778-10785 [Abstract] [Full Text]  
  • Ju, K.-S., Parales, R. E. (2006). Control of substrate specificity by active-site residues in nitrobenzene dioxygenase.. Appl. Environ. Microbiol. 72: 1817-1824 [Abstract] [Full Text]  
  • Jeon, C. O., Park, M., Ro, H.-S., Park, W., Madsen, E. L. (2006). The Naphthalene Catabolic (nag) Genes of Polaromonas naphthalenivorans CJ2: Evolutionary Implications for Two Gene Clusters and Novel Regulatory Control. Appl. Environ. Microbiol. 72: 1086-1095 [Abstract] [Full Text]  
  • Shen, X.-H., Jiang, C.-Y., Huang, Y., Liu, Z.-P., Liu, S.-J. (2005). Functional Identification of Novel Genes Involved in the Glutathione-Independent Gentisate Pathway in Corynebacterium glutamicum. Appl. Environ. Microbiol. 71: 3442-3452 [Abstract] [Full Text]  
  • Keenan, B. G., Leungsakul, T., Smets, B. F., Mori, M.-a., Henderson, D. E., Wood, T. K. (2005). Protein Engineering of the Archetypal Nitroarene Dioxygenase of Ralstonia sp. Strain U2 for Activity on Aminonitrotoluenes and Dinitrotoluenes through Alpha-Subunit Residues Leucine 225, Phenylalanine 350, and Glycine 407. J. Bacteriol. 187: 3302-3310 [Abstract] [Full Text]  
  • Dionisi, H. M., Chewning, C. S., Morgan, K. H., Menn, F.-M., Easter, J. P., Sayler, G. S. (2004). Abundance of Dioxygenase Genes Similar to Ralstonia sp. Strain U2 nagAc Is Correlated with Naphthalene Concentrations in Coal Tar-Contaminated Freshwater Sediments. Appl. Environ. Microbiol. 70: 3988-3995 [Abstract] [Full Text]  
  • Rhee, S.-K., Liu, X., Wu, L., Chong, S. C., Wan, X., Zhou, J. (2004). Detection of Genes Involved in Biodegradation and Biotransformation in Microbial Communities by Using 50-Mer Oligonucleotide Microarrays. Appl. Environ. Microbiol. 70: 4303-4317 [Abstract] [Full Text]  
  • Keenan, B. G., Leungsakul, T., Smets, B. F., Wood, T. K. (2004). Saturation Mutagenesis of Burkholderia cepacia R34 2,4-Dinitrotoluene Dioxygenase at DntAc Valine 350 for Synthesizing Nitrohydroquinone, Methylhydroquinone, and Methoxyhydroquinone. Appl. Environ. Microbiol. 70: 3222-3231 [Abstract] [Full Text]  
  • Miyazawa, D., Mukerjee-Dhar, G., Shimura, M., Hatta, T., Kimbara, K. (2004). Genes for Mn(II)-dependent NahC and Fe(II)-dependent NahH located in close proximity in the thermophilic naphthalene and PCB degrader, Bacillus sp. JF8: cloning and characterization. Microbiology 150: 993-1004 [Abstract] [Full Text]  
  • Ishiyama, D., Vujaklija, D., Davies, J. (2004). Novel Pathway of Salicylate Degradation by Streptomyces sp. Strain WA46. Appl. Environ. Microbiol. 70: 1297-1306 [Abstract] [Full Text]  
  • Zhuang, Z., Song, F., Takami, H., Dunaway-Mariano, D. (2004). The BH1999 Protein of Bacillus halodurans C-125 Is Gentisyl-Coenzyme A Thioesterase. J. Bacteriol. 186: 393-399 [Abstract] [Full Text]  
  • Jeon, C. O., Park, W., Ghiorse, W. C., Madsen, E. L. (2004). Polaromonas naphthalenivorans sp. nov., a naphthalene-degrading bacterium from naphthalene-contaminated sediment. Int. J. Syst. Evol. Microbiol. 54: 93-97 [Abstract] [Full Text]  
  • Van Hamme, J. D., Singh, A., Ward, O. P. (2003). Recent Advances in Petroleum Microbiology. Microbiol. Mol. Biol. Rev. 67: 503-549 [Abstract] [Full Text]  
  • Kasai, Y., Shindo, K., Harayama, S., Misawa, N. (2003). Molecular Characterization and Substrate Preference of a Polycyclic Aromatic Hydrocarbon Dioxygenase from Cycloclasticus sp. Strain A5. Appl. Environ. Microbiol. 69: 6688-6697 [Abstract] [Full Text]  
  • Jones, R. M., Britt-Compton, B., Williams, P. A. (2003). The Naphthalene Catabolic (nag) Genes of Ralstonia sp. Strain U2 Are an Operon That Is Regulated by NagR, a LysR-Type Transcriptional Regulator. J. Bacteriol. 185: 5847-5853 [Abstract] [Full Text]  
  • Chang, H.-K., Mohseni, P., Zylstra, G. J. (2003). Characterization and Regulation of the Genes for a Novel Anthranilate 1,2-Dioxygenase from Burkholderia cepacia DBO1. J. Bacteriol. 185: 5871-5881 [Abstract] [Full Text]  
  • Lessner, D. J., Parales, R. E., Narayan, S., Gibson, D. T. (2003). Expression of the Nitroarene Dioxygenase Genes in Comamonas sp. Strain JS765 and Acidovorax sp. Strain JS42 Is Induced by Multiple Aromatic Compounds. J. Bacteriol. 185: 3895-3904 [Abstract] [Full Text]  
  • Baldwin, B. R., Nakatsu, C. H., Nies, L. (2003). Detection and Enumeration of Aromatic Oxygenase Genes by Multiplex and Real-Time PCR. Appl. Environ. Microbiol. 69: 3350-3358 [Abstract] [Full Text]  
  • Wilson, M. S., Herrick, J. B., Jeon, C. O., Hinman, D. E., Madsen, E. L. (2003). Horizontal Transfer of phnAc Dioxygenase Genes within One of Two Phenotypically and Genotypically Distinctive Naphthalene-Degrading Guilds from Adjacent Soil Environments. Appl. Environ. Microbiol. 69: 2172-2181 [Abstract] [Full Text]  
  • Peng, X., Misawa, N., Harayama, S. (2003). Isolation and Characterization of Thermophilic Bacilli Degrading Cinnamic, 4-Coumaric, and Ferulic Acids. Appl. Environ. Microbiol. 69: 1417-1427 [Abstract] [Full Text]  
  • Parales, R. E., Bruce, N. C., Schmid, A., Wackett, L. P. (2002). Biodegradation, Biotransformation, and Biocatalysis (B3). Appl. Environ. Microbiol. 68: 4699-4709 [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]  
  • Park, W., Padmanabhan, P., Padmanabhan, S., Zylstra, G. J., Madsen, E. L. (2002). nahR, encoding a LysR-type transcriptional regulator, is highly conserved among naphthalene-degrading bacteria isolated from a coal tar waste-contaminated site and in extracted community DNA. Microbiology 148: 2319-2329 [Abstract] [Full Text]  
  • Lessner, D. J., Johnson, G. R., Parales, R. E., Spain, J. C., Gibson, D. T. (2002). Molecular Characterization and Substrate Specificity of Nitrobenzene Dioxygenase from Comamonas sp. Strain JS765. Appl. Environ. Microbiol. 68: 634-641 [Abstract] [Full Text]  
  • Ferrero, M., Llobet-Brossa, E., Lalucat, J., Garcia-Valdes, E., Rossello-Mora, R., Bosch, R. (2002). Coexistence of Two Distinct Copies of Naphthalene Degradation Genes in Pseudomonas Strains Isolated from the Western Mediterranean Region. Appl. Environ. Microbiol. 68: 957-962 [Abstract] [Full Text]  
  • Hickey, W. J., Sabat, G., Yuroff, A. S., Arment, A. R., Perez-Lesher, J. (2001). Cloning, Nucleotide Sequencing, and Functional Analysis of a Novel, Mobile Cluster of Biodegradation Genes from Pseudomonas aeruginosa Strain JB2. Appl. Environ. Microbiol. 67: 4603-4609 [Abstract] [Full Text]  
  • Khan, A. A., Wang, R.-F., Cao, W.-W., Doerge, D. R., Wennerstrom, D., Cerniglia, C. E. (2001). Molecular Cloning, Nucleotide Sequence, and Expression of Genes Encoding a Polycyclic Aromatic Ring Dioxygenase from Mycobacterium sp. Strain PYR-1. Appl. Environ. Microbiol. 67: 3577-3585 [Abstract] [Full Text]  
  • Zhou, N.-Y., Fuenmayor, S. L., Williams, P. A. (2001). nag Genes of Ralstonia (Formerly Pseudomonas) sp. Strain U2 Encoding Enzymes for Gentisate Catabolism. J. Bacteriol. 183: 700-708 [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]  
  • Dunwell, J. M., Khuri, S., Gane, P. J. (2000). Microbial Relatives of the Seed Storage Proteins of Higher Plants: Conservation of Structure and Diversification of Function during Evolution of the Cupin Superfamily. Microbiol. Mol. Biol. Rev. 64: 153-179 [Abstract] [Full Text]  
  • Miyauchi, K., Adachi, Y., Nagata, Y., Takagi, M. (1999). Cloning and Sequencing of a Novel meta-Cleavage Dioxygenase Gene Whose Product Is Involved in Degradation of gamma -Hexachlorocyclohexane in Sphingomonas paucimobilis. J. Bacteriol. 181: 6712-6719 [Abstract] [Full Text]  
  • Tsoi, T. V., Plotnikova, E. G., Cole, J. R., Guerin, W. F., Bagdasarian, M., Tiedje, J. M. (1999). Cloning, Expression, and Nucleotide Sequence of the Pseudomonas aeruginosa 142 ohb Genes Coding for Oxygenolytic ortho Dehalogenation of Halobenzoates. Appl. Environ. Microbiol. 65: 2151-2162 [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]  
  • Laurie, A. D., Lloyd-Jones, G. (1999). The phn Genes of Burkholderia sp. Strain RP007 Constitute a Divergent Gene Cluster for Polycyclic Aromatic Hydrocarbon Catabolism. J. Bacteriol. 181: 531-540 [Abstract] [Full Text]  
  • Wilson, M. S., Bakermans, C., Madsen, E. L. (1999). In Situ, Real-Time Catabolic Gene Expression: Extraction and Characterization of Naphthalene Dioxygenase mRNA Transcripts from Groundwater. Appl. Environ. Microbiol. 65: 80-87 [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»