PURIFICATION OF PIG-HEART BENZYLAMINE OXIDASE

A benzylamine oxidase (E.C. 1.4.3.6) has been purified from pig heart. Western blot analysis showed that the enzyme cross-reacts with a poly clonal antibody raised against homogeneous, crystalline pig plasma ben zylamine oxidase (BAO). A subunit molecular mass of 97 kDa obtained by SDS electrophoresis is identical to the plasma enzyme. The purificati on procedure consisted of sequential DEAE-cellulose, DEAE-Sephadex, Co n A-Sepharose, Sephadex G 200 and hydroxyapatite columns. The specific activity of the purified enzyme was 0.037 mu mol min(-1)mg(-1) at 37 degrees C and the K-m for benzylamine was estimated to be 29 mu M The enzyme was inhibited by carbonyl reagents such as semicarbazide and al pha-aminoguanidine. Phenylhydrazine reacts mole to mole with the enzym e giving a peak at 425 nm. The copper content was 2 g-atom/mole of enz yme.