A simple and rapid high recovery protocol for the purification of arginase

Summary A protocol of purification is devised that gave high recovery of homogeneous arginase from ox erythrocytes. The protocol involved haemolysis, heat treatment, CM- and DEAE-Sepharose chromatography, arginine AH-Sepharose chromatography and molecular sieving through Biogel P-150, all in the presence of 2-mercaptoethanol. It afforded (a) elution of arginase as a single peak, (b) almost 2900 fold purification, (c) 30% recovery and (d) electrophoretically homogeneous arginase. The purified arginase exhibited (a) M.W. of 115,000, (b) dissociation upon SDS treatment into homologous monomers of 30 kDa, (c) optimum pH and temperature 11.5 and 55° C, respectively and (d) Km value for L-arginine-HCl 2.2–2.3 mM, characteristic of the ureotelic type.