[26] Yeast mitochondria and submitochondrial particles

摘要: Publisher Summary Mitochondria and submitochondrial particles have been prepared from several species of yeast, including Candida (Torulopsis) utilis , Saccharomyces cerevisiae closely related such as S. carlsbergensis . The foremost experimental obstacle in the preparation intact yeast mitochondria is need for efficiently breaking refractory cell wall without extensively damaging liberated mitochondria. Two approaches to this problem used: (1) Cells are ruptured mechanically mitochondrial fragments separated more by differential centrifugation careful separation supernatants loosely packed (fluffy) layers (submitochondrial particles) firmer pellets (mitochondria). (2) Yeast cells subjected enzymatic digestion order form osmotically sensitive spheroplasts (protoplasts), which osmotic shock mild homogenization. Several mechanical devices employed various shakers, French pressure cell, blendors with overhead drive, certain types mills. All these under appropriate conditions yielded or capable oxidative phosphorylation, but only a few instances has acceptor control respiration demonstrated preparations. procedures described chapter employ colloid mill; one method yields control.