[Enzymologic characterization of adenosine nucleosidase of medicinal plants (Medicago sativa)]

Adenosine nucleosidase (adenosine ribohydrolase, E C 3.2.2.7) was purified from alfalfa leaf juice. The final preparation shows a single band on polyacrylamide gel electrophoresis; the enzyme activity is stable for 12 hrs between pH 5.5 and pH 8.5, but is completely lost on heating at 55 degrees C for 10 min. Optimal pH for the hydrolysis of adenosine is between pH 5 and pH 6. Among nine purine nucleosides tested, only adenosine, 2'-deoxyadenosine and purine riboside were hydrolyzed by the enzyme preparation. A Km value of 7 x 10(-6) M was found with adenosine as substrate at pH 7.4. Of the two reaction products, adenine exerted a weak inhibitory effect, while D-ribose was without effect on the initial rate of adenosine hydrolysis. The data reported are compared with those obtained on the enzymes from other plant sources.