Cloning, Expression, and Characterization of the katG Gene, Encoding Catalase-Peroxidase, from the Polycyclic Aromatic Hydrocarbon-Degrading Bacterium Mycobacterium sp. Strain PYR-1
- 1 October 2000
- journal article
- research article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 66 (10), 4300-4304
- https://doi.org/10.1128/aem.66.10.4300-4304.2000
Abstract
A 81-kDa protein from Mycobacterium sp. strain PYR-1 was expressed in response to exposure of the strain to the polycyclic aromatic hydrocarbon pyrene and recovered by two-dimensional gel electrophoresis. The N-terminal sequence of the protein indicated that it was similar to catalase-peroxidase. An oligonucleotide probe designed from this sequence was used to screen a genomic library of Mycobacterium sp. strain PYR-1, and a positive clone, containing a part of the gene encoding the 81-kDa protein, was isolated. A gene-walking technique was used to sequence the entire gene, which was identified as katG for catalase-peroxidase. The deduced KatG protein sequence showed significant homology to KatGII of Mycobacterium fortuitum and clustered with catalase-peroxidase proteins from other Mycobacterium species in a phylogenetic tree. The katG gene was expressed in Escherichia coli to produce a protein with catalase-peroxidase activity. Since the induction of this catalase-peroxidase occurred in pyrene-induced cultures and the exposure of these cultures to hydrogen peroxide reduced pyrene metabolism, our data suggest that this enzyme plays a role in polycyclic aromatic hydrocarbon metabolism by strain PYR-1.Keywords
This publication has 16 references indexed in Scilit:
- Gapped BLAST and PSI-BLAST: a new generation of protein database search programsNucleic Acids Research, 1997
- Bioremediation of environments contaminated by polycyclic aromatic hydrocarbonsPublished by Cambridge University Press (CUP) ,1996
- PCR Detection of Polycyclic Aromatic Hydrocarbon-Degrading MycobacteriaEnvironmental Science & Technology, 1995
- Hydrogen peroxide-mediated inactivation of microsomal cytochrome P450 during monooxygenase reactionsFree Radical Biology & Medicine, 1994
- Biodegradation of polycyclic aromatic hydrocarbonsCurrent Opinion in Biotechnology, 1993
- Bioremediation of soil contaminated with polynuclear aromatic hydrocarbons (PAHs): A reviewEnvironmental Pollution, 1993
- The catalase—peroxidase gene and isoniazid resistance of Mycobacterium tuberculosisNature, 1992
- ES&T Special Report: Priority pollutants: I-a perspective viewEnvironmental Science & Technology, 1979
- A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye BindingAnalytical Biochemistry, 1976
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976