Rat kidney contains an enzyme which hydrolyzes NG-monomethyl-L-arginine to give rise to the formation of ornithine and N-methylurea. Confirmation of formation of these reaction products is carried out colorimetrically as well as radiochemically employing NG-monomethyl-L-[ornithine-14C(U)]arginine. This pattern of reaction products suggests that the enzyme responsible is an arginase type. However, the kidney enzyme is quite distinct from the hepatic arginase; commercial bovine hepatic arginase (L-arginine amidinohydrolase, EC 3.5.3.1) is completely inactive toward NG-monomethyl-L-arginine. Among various rat tissues examined, the hydrolytic activity is the highest in kidney, followed by the activity in liver and pancreas.