Purification of Aconitase

The aconitase activity staining method on the polyacrylamide gel disc electrophoresis was devised and applied to the detection of aconitase. Aconitase of swine heart was purified by ammonium sulfate fractionation and column chromatographies. By chromatography on hydroxy-lapatite or by polyacrylamide gel electrophoresis, the ammoniun sulfate fraction of swine heart aconitase was separated into two forms. The elution volumes of both aconitase fractions were equal by column chromatography on Sephadex G-200. One form of aconitase of ammonium sulfate fraction was obtained by column chromatography on DEAE-Sephadex A-50. This form was rechromatographed on CM-Sephadex C-50 and was found to be fairly homogeneous by disc electrophoresis.