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Antimicrobial Agents and Chemotherapy, April 2003, p. 1343-1346, Vol. 47, No. 4
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.4.1343-1346.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Chromosomal aadD2 Encodes an Aminoglycoside Nucleotidyltransferase in Bacillus clausii

Bülent Bozdogan,1,{dagger} Sébastien Galopin,1 Guy Gerbaud,2 Patrice Courvalin,2 and Roland Leclercq1*

Service de Microbiologie, CHU Côte de Nacre, Caen,1 Unité des Agents Antibactériens, Institut Pasteur, Paris, France2

Received 26 August 2002/ Returned for modification 15 November 2002/ Accepted 18 January 2003

Bacillus clausii SIN is one of the four strains of B. clausii composing a probiotic administered to humans for the prevention of gastrointestinal side effects due to oral antibiotic therapy. The strain is resistant to kanamycin, tobramycin, and amikacin. A gene conferring aminoglycoside resistance was cloned into Escherichia coli and sequenced. The gene, called aadD2, encoding a putative 246-amino acid protein, shared 47% identity with ant(4')-Ia from Staphylococcus aureus, which encodes an aminoglycoside 4'-O-nucleotidyltransferase. Phosphocellulose paper-binding assays indicated that the gene product was responsible for nucleotidylation of kanamycin, tobramycin, and amikacin. The aadD2 gene was detected by DNA-DNA hybridization in the three other strains of the probiotic mixture and in the reference strain B. clausii DSM8716, although it did not confer resistance in these strains. Mutations in the sequence of the putative promoter for aadD2 from B. clausii SIN resulted in higher identity with consensus promoter sequences and may account for aminoglycoside resistance in that strain. The aadD2 gene was chromosomally located in all strains and was not transferable by conjugation. These data indicate that chromosomal aadD2 is specific to B. clausii.

* Corresponding author. Mailing address: Service de Microbiologie, CHU de Caen, Avenue de la Côte de Nacre, 14033 Caen Cedex, France. Phone: (33) 2 31 06 48 95. Fax: (33) 2 31 06 45 73. E-mail: leclercq-r{at}chu-caen.fr.

{dagger} Present address: Department of Pathology, Hershey Medical Center, Hershey, PA 17033.

Antimicrobial Agents and Chemotherapy, April 2003, p. 1343-1346, Vol. 47, No. 4
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.4.1343-1346.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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