Overexpression and Initial Characterization of the Chromosomal Aminoglycoside 3′-O-Phosphotransferase APH(3′)-IIb from Pseudomonas aeruginosa

ABSTRACT 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 × 104 to 36 × 104 M−1 s−1.

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