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A coupled optical enzyme assay for phosphopentomutase
Authors:Maria Grazia Tozzi  Roberta Catalani  Pier Luigi Ipata  Umberto Mura
Institution:Institute of Biochemistry, Biophysics and Genetics, Faculty of Science, University of Pisa, Via A. Volta 4, 56100 Pisa, Italy
Abstract:Published assays for phosphopentomutase activity are based on acid lability differences between ribose 1-phosphate and ribose 5-phosphate. The present work describes a new method in which the isomerization of ribose 5-phosphate to ribose 1-phosphate is followed spectrophotometrically at 265 nm by coupling it with the following two-stage enzymatic conversion: ribose 1-phosphate + adenine ? phosphate + adenosine (adenosine phosphorylase); adenosine + H2O → inosine + NH3 (adenosine deaminase). The method has been used to show some properties of Escherichia coli phosphopentomutase.
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