Purification of peroxidases and separation of isoperoxidases from various plant species

Abstract

The partial purification of peroxidase (EC.1.11.1.7) and separation of isoperoxidases by disc electrophoresis from Cucurbita Pepo L., Phaseolus vulgaris L., Cicer arietinum L. and Hordeum, Secale and Triticum sp., have been studied. Peroxidase from fruit of pumpkin and from 6-day-old coleoptiles of French bean and chick pea has been partially purified, 128-, 174-, and 140-fold, respectively. The apparent Km at the optimum pH were: pumpkin (epicarp.), 2.7 X 10(-4) M; barley, common rye and wheat (primary leaves, in all cases), 1.4 X 10(-5), 1.2 X 10(-5) and 3.1 X 10(-5) M, respectively. Isoperoxidases have been separated by disc electrophoresis on 7% polyacrylamide gel and stained with p-phenylenediamine. Differences in patterns of anodic and cathodic isozymes were observed: 3 isozymes from fruits of pumpkin, 4 from French bean, 4 from chick pea, 11 from leaves of barley, 10 from leaves of common rye and 9 from leaves of wheat.