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 Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Iwabuchi, T. Articles by Harayama, S. Search for Related Content PubMed PubMed Citation Articles by Iwabuchi, T. Articles by Harayama, S.

 Previous Article  |  Next Article 

J. Bacteriol., Oct 1997, 6488-6494, Vol 179, No. 20
Copyright © 1997, American Society for Microbiology

Biochemical and genetic characterization of 2-carboxybenzaldehyde dehydrogenase, an enzyme involved in phenanthrene degradation by Nocardioides sp. strain KP7

T Iwabuchi and S Harayama
Marine Biotechnology Institute, Kamaishi Laboratories, Iwate, Japan.

2-Carboxybenzaldehyde dehydrogenase from the phenanthrene-degrading bacterium Nocardioides sp. strain KP7 was purified and characterized. The purified enzyme had a molecular mass of 53 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 205 kDa by gel filtration chromatography. Thus, the homotetramer of the 53-kDa subunit constituted an active enzyme. The apparent Km and kcat values of this enzyme for 2-carboxybenzaldehyde were 100 microM and 39 s(-1), respectively, and those for NAD+ were 83 microM and 32 s(-1), respectively. The structural gene for this enzyme was cloned and sequenced. The length of the gene was 1,455 bp. The nucleotide sequence of the 10,279 bp of DNA around the gene for 2-carboxybenzaldehyde dehydrogenase was also determined, and seven open reading frames were found in this DNA region. These were the genes for 1-hydroxy-2- naphthoate dioxygenase (phdI) and trans-2'-carboxybenzalpyruvate aldolase (phdJ), orf1, the gene for 2-carboxybenzaldehyde dehydrogenase (phdK), orf2/orf3, and orf4. The amino acid sequence of the orf1 product was similar to that of the aromatic hydrocarbon transporter gene (pcaK) in Pseudomonas putida PRS2000. The amino acid sequence of the orf4 product revealed a similarity to cytochrome P-450 proteins. The region between phdK and orf4 encoded orf2 and orf3 on different strands. The amino acid sequences of the orf2 and orf3 products exhibited no significant similarity to the reported sequences in protein databases.

This article has been cited by other articles:

• 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]  
• Chauvaux, S., Chevalier, F., Le Dantec, C., Fayolle, F., Miras, I., Kunst, F., Beguin, P. (2001). Cloning of a Genetically Unstable Cytochrome P-450 Gene Cluster Involved in Degradation of the Pollutant Ethyl tert-Butyl Ether by Rhodococcus ruber. J. Bacteriol. 183: 6551-6557 [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]  
• Segura, A., Bünz, P. V., D'Argenio, D. A., Ornston, L. N. (1999). Genetic Analysis of a Chromosomal Region Containing vanA and vanB, Genes Required for Conversion of Either Ferulate or Vanillate to Protocatechuate in Acinetobacter. J. Bacteriol. 181: 3494-3504 [Abstract] [Full Text]  
• Adachi, K., Iwabuchi, T., Sano, H., Harayama, S. (1999). Structure of the Ring Cleavage Product of 1-Hydroxy-2-Naphthoate, an Intermediate of the Phenanthrene-Degradative Pathway of Nocardioides sp. Strain KP7. J. Bacteriol. 181: 757-763 [Abstract] [Full Text]  
• Werwath, J., Arfmann, H.-A., Pieper, D. H., Timmis, K. N., Wittich, R.-M. (1998). Biochemical and Genetic Characterization of a Gentisate 1,2-Dioxygenase from Sphingomonas sp. Strain RW5. J. Bacteriol. 180: 4171-4176 [Abstract] [Full Text]  
• Iwabuchi, T., Harayama, S. (1998). Biochemical and Molecular Characterization of 1-Hydroxy-2-naphthoate Dioxygenase from Nocardioides sp. KP7. J. Biol. Chem. 273: 8332-8336 [Abstract] [Full Text]  
• Iwabuchi, T., Harayama, S. (1998). Biochemical and Genetic Characterization of trans-2'-Carboxybenzalpyruvate Hydratase-Aldolase from a Phenanthrene-Degrading Nocardioides Strain. J. Bacteriol. 180: 945-949 [Abstract] [Full Text]