[Distribution of microbody enzymes from Chlamydomonas on sucrose gradients]

The crude homogenate of cells from Chlamydomonas reinhardii was placed on a linear gradient from 30% to 60% sucrose and centrifuged for 4 hours at 60 000 g. After fractionation of the gradient the distribution of enzymes was determined. Hydroxypyruvate reductase and glycolate dehydrogenase, two markers for peroxisomes, appeared with one sharp peak at density 1.185 g/cm(3) within the gradient. Twenty-five percent of the hydroxypyruvate reductase was particulate and 75% was found as solubles in the top fractions. The peaks of both enzymes matched exactly the peak of the cytochrome oxidase which is a marker for mitochondria. The profile for malate dehydrogenase was the same as that for hydroxypyruvate reductase. Catalase, however, showed two peaks. One coincided with the peak of cytochrome oxidase and the other appeared at density 1.22 g/cm(3). More than 50% of the catalase moved into the gradient during centrifugation.