Journal of Bacteriology, January 2003, p. 135-141, Vol. 185, No. 1
0021-9193/03/$08.00+0 DOI: 10.1128/JB.185.1.135-141.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
Arsenite Oxidase aox Genes from a Metal-Resistant ß-Proteobacterium
Daniel Muller,1 Didier Lièvremont,1 Diliana Dancheva Simeonova,1,2 Jean-Claude Hubert,1 and Marie-Claire Lett1*
Laboratoire de Dynamique, Évolution et Expression des Génomes de Micro-Organismes, FRE2326 Université Louis-Pasteur/CNRS, 67083 Strasbourg cedex, France,1
Laboratory of Geomicrobiology, The Sofia University "St. Kliment Ohridski," 1421 Sofia, Bulgaria2
Received 31 May 2002/
Accepted 9 October 2002
The ß-proteobacterial strain ULPAs1, isolated from an arsenic-contaminated environment, is able to efficiently oxidize arsenite [As(III)] to arsenate [As(V)]. Mutagenesis with a lacZ-based reporter transposon yielded two knockout derivatives deficient in arsenite oxidation. Sequence analysis of the DNA flanking the transposon insertions in the two mutants identified two adjacent open reading frames, named aoxA and aoxB, as well as a putative promoter upstream of the aoxA gene. Reverse transcription-PCR data indicated that these genes are organized in an operonic structure. The proteins encoded by aoxA and aoxB share 64 and 72% identity with the small Rieske subunit and the large subunit of the purified and crystallized arsenite oxidase of Alcaligenes faecalis, respectively (P. J. Ellis, T. Conrads, R. Hille, and P. Kuhn, Structure [Cambridge] 9:125-132, 2001). Importantly, almost all amino acids involved in cofactor interactions in both subunits of the A. faecalis enzyme were conserved in the corresponding sequences of strain ULPAs1. An additional Tat (twin-arginine translocation) signal peptide sequence was detected at the N terminus of the protein encoded by aoxA, strongly suggesting that the Tat pathway is involved in the translocation of the arsenite oxidase to its known periplasmic location.
* Corresponding author. Mailing address: Laboratoire de Microbiologie, FRE 2326 Université Louis-Pasteur/CNRS, 28 rue Goethe, 67000 Strasbourg, France. Phone: (33) 3 90 24 19 97. Fax: (33) 3 90 24 20 28. E-mail: lett{at}gem.u-strasbg.fr.
Journal of Bacteriology, January 2003, p. 135-141, Vol. 185, No. 1
0021-9193/03/$08.00+0 DOI: 10.1128/JB.185.1.135-141.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
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Copyright © 2003 by the American Society for Microbiology. All rights reserved.