Peroxidase extractive purification from plant raw material in aqueous two-phase systems

M.V. MIRANDA; H.M. FERNÁNDEZ LAHORE; J. DOBRECKY; O. CASCONE

ACTA BIOTECHNOLOGICA

WILEY-V C H VERLAG GMBH

Año: 1998 vol. 18 p. 179 - 188

0138-4988

The extractive purification of peroxidase from Armoracia rusficana roots and Glycine mnx seed coats in temperature-induced and affinity microsphere-containing aqueous two-phase systems was studied. The extractive purification of peroxidase from Glycine max seed coats was carried out in a temperature-induced aqueous two-phase system formed by Triton X-45, Triton X-100 and sodium acetate at pH 5.5. A 99% yield with a 6-fold purification factor was obtained. When the clear top phase was subiected to concanavalin-A attnity chromatography, the purification factor rose to 41 and the yield dropped to 28%. A two-step purification process for peroxidase from Armoracia rusticana roots was developed by adding concanavalin-A affinity microspheres to a PEG/phosphate aqueous two-phase system. The method allows a 60% recovery of high purity peroxidase (1,860 guaiacol units per mg). A lower recovery rate and degree of purification of this enzyme was achieved after temperature-induced aqueous two-phase partition or acetone precipitation and concanavalin-A affinity column chromatography.