Purification of Peroxidase from Chinese Cabbage Roots by the Reverse Micelle System

The basic and optimum conditions for the extraction of peroxidase from Chinese cabbage root applying the reverse micelle system were investigated. In order to purify Peroxidase (POX) from crude extract of Chinese cabbage roots, isooctane containing 110 mM Aerosol OT (AOT) was well mixed with the same volume of crude extracts containing 50 mM NaCl and 30 mM Tris-HCI buffer of pH 8.0. After centrifugation, AOT reverse micelle containing the isooctane phase were mixed with 80 mM Tris-HCI buffer at pH 7.0 containing 1 M KCl. From these operations, POX was purified 20-fold with a 60% yield. For further purification, DEAE-Toyopearl column chromatography was applied, and it showed a single protein band on SDS-polyacrylamide gel electrophoresis. The resulting POX showed 93-fold purification with a 40% yield.