Abstract: A novel cyclic nucleotide phosphodiesterase, which is different from known isozyme families, is isolated and purified from rat cerebrum. This enzyme has the following physicochemical properties (1) to (3). (1) The enzyme acts on cAMP to form 5'-AMP and on cGMP to form 5'-GMP; (2) The Km values for cAMP and cGMP are 0.11 .mu.M and 1.78 .mu.M, respectively, as indicating substrate specificity; and (3) the molecular weight is approximately 298000.