Urealytic activity of the cytoplasmic fraction of prepared by digitonin lysis was assayed in a simple buffer system (HEPES plus EDTA) by measuring the release of CO from [C]urea. The of this preparation agreed with our previous observations of the same activity measured in a more complex reaction mixture. The substrate concentration at which maximum velocity occurred was approximately 20 mM. The activity was sensitive to heavy metals and inhibitors which react with sulphydryl groups such as -ethylmaleimide and -chloromercuribenzoate. It was not inhibited by Ca or Mg or by the reaction products, ammonia and carbon dioxide.


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