On the Physiological Significance of the Isoenzyme Groups of Peroxidase from Tabacco Demonstrated by Biochemical Properties I. Separation, Purification, Chemical and Physical Properties

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1977

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Abstract

Separation and purification of three peroxidase-isoenzyme groups (GI, fast migrating anodic; GH, slow migrating anodic; GIII, slow migrating cathodic) was accomplished by ion-exchange chromatography. Chromatographic separation of the groups was controlled by disk-electrophoresis and by isoelectric focusing. Molecular weights were estimated by SDS-gelelectrophoresis and by thin layer chromatography. The results of both methods are similar. GI has a molecular weight of about 27,000, GH of 43,000 and GIII of 47,000. By affinity chromatography with Coocanavalin A-sepharose it was shown that the enzymes of the three peroxidase groups are all glycoproteins. While heat inactivation of GI was measured at 50 °C, inactivation of GII and GIII did not take place before 70 °C. Absorption spectra of the three groups were typical for ferriperoxidases with the maximum of the Soret band at 401 nm.

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peroxidase, izoenzymes, biochemical characterization, callus cultures, nicotiana tabacum

Citation

Z.Pflanzenphysiol.Bd.82.S, 235-246, 1977

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