Purification of peroxidase from Amsonia orientalis by three-phase partitioning and its biochemical characterization

The present work describes the purification and characterization of peroxidase from the medicinal plant, Amsonia orientalis, for the first time. The activity recovery for peroxidase was 162% with 12.5-fold purification. Optimal purification parameters were 20% (w/v) (NH4)(2)SO4 saturation at pH 6.0 and 25 degrees C with 1.0:1.0 (v/v) ratio of crude extract to t-butanol ratio for 30 min. The molecular mass of the enzyme was found to be ca. 59 kDa. Peroxidase showed K-m values of 1.88 and 2.0 mM for pyrogallol and hydrogen peroxide, respectively. FeSO4, CuSO4, HgCl2, MnSO4 and MgSO4 did not inhibit the enzyme activity.