Double-Osmotic-Shock Procedure to Prepare Gentamicin Adenine Mononucleotide Transferase for the Enzymatic Assay of Gentamicin

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Double-osmotic-shock procedure to prepare gentamicin adenine mononucleotide transferase for the enzymatic assay of gentamicin.

The release of gentamicin adenine mononucleotide transferase during single, cold, osmotic-shock treatment of Escherichia coli K-12 W677/HJR66 is not always maximal. The quantity of gentamicin adenine mononucleotide transferase in osmotic extracts could be significantly increased when a double-shock procedure was performed.

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An enzymatic, radiochemical assay procedure for measuring serum gentamicin by use of gentamicin 3-acetyl-transferase and [acetyl-14C]acetyl-Coenzyme A is described and evaluated. The enzyme stoichiometrically and quantitatively transfers a radioactive label to the analyte during a 10-min incubation at 37 degrees C. The labeled gentamicin is then absorbed onto phosphocellulose paper discs, which...

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ژورنال

عنوان ژورنال: Antimicrobial Agents and Chemotherapy

سال: 1977

ISSN: 0066-4804,1098-6596

DOI: 10.1128/aac.11.6.1074