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Applied and Environmental Microbiology, February 2002, p. 634-641, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.2.634-641.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Molecular Characterization and Substrate Specificity of Nitrobenzene Dioxygenase from Comamonas sp. Strain JS765

Daniel J. Lessner,1* Glenn R. Johnson,2 Rebecca E. Parales,1 Jim C. Spain,2 and David T. Gibson1

Department of Microbiology and Center for Biocatalysis and Bioprocessing, The University of Iowa, Iowa City, Iowa 52242,1 Air Force Research Laboratory, Tyndall Air Force Base, Florida 324032

Received 16 July 2001/ Accepted 23 October 2001

Comamonas sp. strain JS765 can grow with nitrobenzene as the sole source of carbon, nitrogen, and energy. We report here the sequence of the genes encoding nitrobenzene dioxygenase (NBDO), which catalyzes the first step in the degradation of nitrobenzene by strain JS765. The components of NBDO were designated ReductaseNBZ, FerredoxinNBZ, OxygenaseNBZ{alpha}, and OxygenaseNBZß, with the gene designations nbzAa, nbzAb, nbzAc, and nbzAd, respectively. Sequence analysis showed that the components of NBDO have a high level of homology with the naphthalene family of Rieske nonheme iron oxygenases, in particular, 2-nitrotoluene dioxygenase from Pseudomonas sp. strain JS42. The enzyme oxidizes a wide range of substrates, and relative reaction rates with partially purified OxygenaseNBZ revealed a preference for 3-nitrotoluene, which was shown to be a growth substrate for JS765. NBDO is the first member of the naphthalene family of Rieske nonheme iron oxygenases reported to oxidize all of the isomers of mono- and dinitrotoluenes with the concomitant release of nitrite.

* Corresponding author. Mailing address: Department of Microbiology, 3-730 BSB, The University of Iowa, Iowa City, IA 52242. Phone: (319) 335-7982. Fax: (319) 335-9999. E-mail: daniel-lessner{at}uiowa.edu.

Applied and Environmental Microbiology, February 2002, p. 634-641, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.2.634-641.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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