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Isolation And Thermodynamic Characterization Of Aminoglycoside Nucleotidyltransferase (2″)-Ia, Roger Edward Wright

Doctoral Dissertations

Aminoglycoside nucleotidyltransferase(2″)-Ia is one of the most important aminoglycoside-modifying enzymes. Because of the difficulties in isolating this enzyme with a high level of purity, very little work has been reported for this enzyme. A procedure for obtaining this enzyme with a purity level of greater than 95% from inclusion bodies was developed. The optimal conditions for isolating ANT(2″) were solubilizing the inclusion bodies in 8 M urea followed by direct dilution of the enzyme into 0.1 M Tris-HCl pH 8.5, 0.2 M KCl, 0.4 ML-arginine and 5 mM reduced glutathione at 4°C. The determination of an effective method of obtaining …