Properties of catalase purified from a methanol‐grown yeast, Kloeckera sp. 2201

Abstract

Catalase, a marker enzyme of peroxisomes, was purified to homogeneity from whole cells of Kloeckera sp.2201 (a strain of Candida boidinii) grown on methanol by means of ammonium sulfate fractionation followed by hydroxyapatite, Sephacryl S‐300 and DEAE‐Sepharose column chromatographies. Crystallized catalase was brown‐coloured and needle‐like. The molecular mass of the enzyme was about 240000 daltons consisting of four identical subunits of 62000 daltons. The minimum size of catalase molecule was estimated to be about 6x10 nm from an electron micrograph. Judging from the absorption spectrum, the enzyme seemed to belong to a group of T‐type catalase. The Km value of the enzyme for hydrogen peroxide (catalatic activity) was 25 mM, while that for methanol (peroxidatic activity) was 83 mM. Catalase from Kloeckera sp. cells showed a certain degree of ‐similarity to the enzyme purified from alkane‐grown Candida tropicalis [T. Yamada et al. (1982) Eur. J. Biochem. 125, 517–521 and 129, 251–255] in its immunochemical properties. Copyright © 1986, Wiley Blackwell. All rights reserved