Summary: Extracts of glucose-grown and methanol-grown were fractionated on sucrose gradients. Mitochondria and microbodies from methanol-grown cells were more fragile than those from glucose-grown cells. Formaldehyde and formate dehydrogenases and antimycin A-insensitive NADPH: cytochrome oxidoreductase were non-sedimentable enzymes. The mitochondrial zone defined by cytochrome oxidase ( = 1·19 to 1·20 g ml) contained sedimentable activities of malate dehydrogenase and citrate synthase. The vacuolar enzyme acid phosphatase was also recovered from this region. The microbody enzyme catalase sedimented to a lower density ( = 1·17 g ml) than mitochondria after growth on glucose but, together with methanol oxidase activity, to higher densities ( = 1·23 and 1·27 g ml) after growth on methanol. Activity of the methanol assimilation enzyme dihydroxyacetone synthase was partially sedimentable and was recovered in the fraction containing mitochondria and membraneous vesicles. Inosine diphosphatase was probably multilocational in the cells.


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