Crystallization and Characterization of Polyamine Oxidase from Penicillium chrysogenum

Abstract

Polyamine oxidase was purified and crystallized with an overall yield of 35% from mycelial extract of Penicillium chrysogenum by a procedure involving ammonium sulfate fractionation, and DEAE-cellulose and Sephadex G-200 column chromatographies. The crystalline enzyme was homogeneous, as judged by disc gel electrophoresis and ultracentrifugation. The sedimentation coefficient (s⁰_{20, w}) of the enzyme was determined to be 6.9S, and diffusion coefficient (D_{20, w}) to be 4.2×10^-7cm² sec^-1. The enzyme showed a molecular weight of about 160, 000 by gel filtration method and ultracentrifugal analysis, and it was composed of two identical subunits. The enzyme was a flavoprotein with absorption maxima at 275, 375 and 450nm. The prosthetic group was identified to be FAD. The enzyme oxidized spermine, and slightly oxidized spermidine. Diamines and monoamines were not oxidized.