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Antimicrobial Agents and Chemotherapy, February 2007, p. 774-776, Vol. 51, No. 2
0066-4804/07/$08.00+0     doi:10.1128/AAC.01034-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Overexpression and Initial Characterization of the Chromosomal Aminoglycoside 3'-O-Phosphotransferase APH(3')-IIb from Pseudomonas aeruginosa{triangledown},{dagger}

Mariana Hainrichson,1 Orit Yaniv,1,2 Marina Cherniavsky,1 Igor Nudelman,1 Dalia Shallom-Shezifi,1 Sima Yaron,2 and Timor Baasov1*

The Edith and Joseph Fischer Enzyme Inhibitors Laboratory, The Schulich Faculty of Chemistry and Institute of Catalysis Science and Technology,1 Department of Biotechnology and Food Engineering, Technion—Israel Institute of Technology, Haifa 32000, Israel2

Received 18 August 2006/ Returned for modification 5 September 2006/ Accepted 24 October 2006

The chromosomal gene aph(3')-IIb, encoding an aminoglycoside 3'-phosphotransferase in Pseudomonas aeruginosa, was cloned and overexpressed in Escherichia coli. The APH(3')-IIb enzyme was purified as a monomer in a two-step procedure and was shown to phosphorylate its substrates at the C-3'-OH position, with kcat/Km values of 0.4 x 104 to 36 x 104 M–1 s–1.

* Corresponding author. Mailing address: The Schulich Faculty of Chemistry, Technion—Israel Institute of Technology, Haifa 32000, Israel. Phone: (972) 4-829-2590. Fax: (972) 4-829-5703. E-mail: chtimor{at}tx.technion.ac.il.

{dagger} Supplemental material for this article may be found at http://aac.asm.org/.

{triangledown} Published ahead of print on 6 November 2006.

Antimicrobial Agents and Chemotherapy, February 2007, p. 774-776, Vol. 51, No. 2
0066-4804/07/$08.00+0     doi:10.1128/AAC.01034-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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