[Isolation of vacuoles from Candida tropicalis]

Intact vacuoles were isolated from the homogenate of the Candida tropicalis protoplasts by flotation in a stepwise gradient of phycoll density. New parameters of protoplast decomposition in hypotonic and isotonic conditions yielding intact vacuoles have been found, as well parameters of density gradient and conditions of centrifugation which differ essentially from parameters used by other authors for isolating the vacuoles from the Saccharomyces cells. The purity of the vacuole preparations was confirmed by using enzymes, light and electron microscopy. The specific content of the carbon of protein, lipids, and free amino acids in the vacuoles and protoplasts of the yeast cells cultivated on 14C-labelled glucose was determined by the radioindicator technique.