ISOLATION AND CHARACTERIZATION OF MICROBODIES FROM CANDIDA TROPICALIS PK 233 CELLS GROWN ON NORMAL ALKANES

Abstract

Numerous microbodies developed at the logarithmic phase of growth in cells of Candida tropicalis, when cultures were grown in a medium containing normal alkanes. From the homogenate of protoplasts prepared from 16-hr cells of Candida, biologically active microbodies were isolated by discontinuous sucrose density gradient centrifugation. The fractions recovered in the middle and lower-density regions of the gradient were associated with substantially high activities of three peroxisomal marker enzymes, catalase, D-amino acid oxidase, and urate oxidase, while a mitochondrial marker enzyme, cytochrome oxidase, was only slightly detected. Electron microscopic studies revealed that these fractions were largely comprised of morphologically well-preserved microbodies with a smaller number of more or less degraded organelles, but were virtually devoid of mitochondria. The microbody fractions contained an appreciable amount of DNA.