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Applied and Environmental Microbiology, October 2000, p. 4300-4304, Vol. 66, No. 10
0099-2240/00/$04.00+0

Cloning, Expression, and Characterization of the katG Gene, Encoding Catalase-Peroxidase, from the Polycyclic Aromatic Hydrocarbon-Degrading Bacterium Mycobacterium sp. Strain PYR-1

Rong-Fu Wang,1 David Wennerstrom,2 Wei-Wen Cao,1 Ashraf A. Khan,1 and Carl E. Cerniglia1,*

Microbiology Division, National Center for Toxicological Research, Food and Drug Administration, Jefferson, Arkansas 72079,1 and Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 722052

Received 2 May 2000/Accepted 4 August 2000

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.

* Corresponding author. Mailing address: Microbiology Division, National Center for Toxicological Research, FDA, Jefferson, AR 72079. Phone: (870) 543-7341. Fax: (870) 543-7307. E-mail: CCerniglia{at}nctr.fda.gov.

Applied and Environmental Microbiology, October 2000, p. 4300-4304, Vol. 66, No. 10
0099-2240/00/$04.00+0


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