Belov A P, Davidova E G, Rachinskiĭ V V
Mikrobiologiia. 1976 Sep-Oct;45(5):852-8.
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.
通过在逐步增加的藻酸盐密度梯度中浮选，从热带假丝酵母原生质体的匀浆中分离出完整的液泡。发现了在低渗和等渗条件下产生完整液泡的原生质体分解的新参数，以及密度梯度参数和离心条件，这些参数与其他作者从酿酒酵母细胞中分离液泡所使用的参数有本质区别。通过酶、光学和电子显微镜确认了液泡制剂的纯度。采用放射性指示剂技术测定了在14C标记葡萄糖上培养的酵母细胞的液泡和原生质体中蛋白质、脂质和游离氨基酸的碳的特定含量。
