Overexpression and initial characterization of the chromosomal aminoglycoside 3'-O-phosphotransferase APH(3')-IIb from Pseudomonas aeruginosa.
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 0.4x10(4) to 36x10(4) M-1 s-1.
Hainrichson
Mariana
M
The Edith and Joseph Fischer Enzyme Inhibitors Laboratory, The Schulich Faculty of Chemistry, Department of Biotechnology and Food Engineering, Technion-Israel Institute of Technology, Haifa 32000, Israel.
Yaniv
Orit
O
Cherniavsky
Marina
M
Nudelman
Igor
I
Shallom-Shezifi
Dalia
D
Yaron
Sima
S
Baasov
Timor
T
D001426:Bacterial Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D019868:Kanamycin Kinase / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D007700:Kinetics; D010766:Phosphorylation; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics
D001426:Bacterial Proteins; D019868:Kanamycin Kinase