Add time:09/06/2019         Source:sciencedirect.com

Publisher SummaryThis chapter discusses the preparation and analysis of ribose-5-phosphate. Two methods of preparation of ribose-5-phosphate are presented in this chapter. One of these is based on the hydrolysis of adenylic acid; the other involves the esterification of free ribose with ATP in the presence of ribokinase and is useful for the preparation of ribose-5-phosphate labeled with C14 or p32. For hydrolysis of adenylic acid one gram of adenosine-5- phosphoric acid is dissolved in 10 ml of 1 N hydrochloric acid and heated for 30 minutes in a boiling water bath. The solution is cooled and centrifuged. The precipitate which forms when the extracts are combined is dissolved by the addition of water, and the solution is treated with of ethanol. The precipitate is collected by centrifugation and dried in vacuo water bath. In the second method, ribokinase is isolated and is assayed in a mixture containing 0.05 ml. of 0.02 M D-ribose, 0.05 ml. of 0.10 M MgCl2, 0.05 ml. of 0.045 M ATP, 0.50 ml. of 0.05 M Tris buffer, pH 7.5, and 0.05 to 0.20 ml. of ribokinase, incubated at 25 °.

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