Biochimie 70 (1988) 1759-1763 (~) Soci6t6de Chimie biologique/Elsevier, Paris 1759 Research article One-step purification and properties of catalase from leaves of Zantedeschia aethiopica Helena TRINDADE 1, Amin KARMALI* and Maria S. PAIS .1 LNETI / DTIQ-Bioqufmica, Estrada das Palmeiras 2745 Queluz de Bai.,:.o; and I Falcudade de Ci#ncias de Lisboa, Departamento de Biologica Vegetal, Edificio C2, Campo Grande 1600, Lisbon, Portugal (Received 25-1-1988, acceptedafter revision 11-5-1988) Summary -- Catalase (E.C 1.11.1.6) was purified from leaves of Zandedeschia aethiopica to apparent homogeneity by a one-step hydrophobic interaction chromatography on a phenyl Sepharose CL-4B column. The purified enzyme preparation was obtained with a final recovery of enzyme activity of about 61% and a specific activity of 146 U/mg protein. The purified enzyme ran as a single protein band when analyzed both by native PAGE and SDS-PAGE corresponding to an Mr of 220,000 Da, which consists of 4 subunits with identical Mr of 54,000 Da. The pI of purified enzyme was found to be 5.2 by isoelectric focusing on ultrathin polyacrylamide gels. The purified catalase has an optimum temperature of activity at 40oC, whereas it is stable between 0 ° and 50oC. As regards pH, the enzyme has an optimum activity at pH 7.0 and it is stable in the range pH 6-8. The absorption spectrum of the purified enzyme exhibited 2 peaks at 280 nm and 405 nm. one-step purification/ catalase / Zantedesehia aethiopica / hydrophobic interaction chromatography/ properties Introduction Catalase (EC i.11.1.6) was first isolated by Chance from bovine liver [1]. Since then~ this enzyme has been found to occur in a number of mammalian tissues, plants, and microorganisms [2-5]. The enzyme from plants and animals is located in microbodies (i.e peroxisomes and glyoxysomes) and often serves as a marker for these cellular organelles [6, 7]. The regreening of Zantedeschia aethiopica fruiting spathe follow- ing the action of endogenous cytokinins is accompanied by peroxisome restructuring and proliferation, as well as by an increase in catalase and glycollate oxidase content of these cellular organelles [8, 9]. Therefore, the availability of a highly purified preparation of catalase from Z. aethiopica would be of interest to study the biogenesis and proliferation of the peroxisomes of this plant. Furthermore, this would permit detailed study of the effect of cytokinins on biogenesis and proliferation of such peroxisomes [8, 91. Catalase has been purified from a number of sources to apparent homogeneity by means of a series of steps involving precipitation either with organic solvents or ammonium sulfate and conventional chromatography, which result in low yields of enzyme activity [10-15]. The present work reports a one-step purification of catalase from Z. aethiopica by hydrophobic inter- action chromatography, and some of its physico- chemical properties are presented. Materials and methods Materials Polyvinylpolypyrrolidone (PVP), bovine serum albumin (BSA), ovalbumin, chymotrypsin, alcohol dehydrogenase (yeast), amyloglucosidase, glucose oxidase, catalase (bovine liver), peroxidase (horsera- dish), diaminobenzidine, bovine liver catalase, *Correspondence and reprints.