Add time:07/22/2019         Source:sciencedirect.com

Publisher SummaryThis chapter discusses the preparation and purification of adenylosuccinase and adenylosuccinic acid. The product of the reversible enzymatic reaction between fumaric acid and adenosine 5′-phosphate, adenylosuccinic acid, is prepared in millimole quantities by use of purified yeast adenylosuccinase. Fumarase was removed from the preparation at this stage by heat inactivation. Ten minutes of beating at 60° resulted in complete inactivation of fumarase with about 90% retention of adenylosuccinase activity. After chilling of the preparation to 4°, denatured protein was removed by centrifugation. The supernatant solution was dialyzed against 0.1 M phosphate buffer, pH 6.5, and this fraction, which was stable for several months at -10°, was employed for the synthesis of adenylosuccinate. One millimole of adenosine 5′-phosphate and 9 millimoles of fumaric acid were dissolved in 50 ml of distilled water by adjusting the pH of the solution to 6.0. It is found that adenylosuccinic acid is hydrolyzed to 6-succinoaminopurine and ribose 5′-phosphate by heating in 1N HCl at 90° for 15 minutes.

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