Pechatnikova I Sh, Efimova T P, Tereshin I M
Mikrobiologiia. 1976 Sep-Oct;45(5):859-63.
Protoplasts of Actinomyces hygroscopicus were obtained in a hypertonic medium containing 10.3% sucrose. Lysozyme was employed as a lysing agent at a concentration of 1 mg/ml. The cells were subjected to lysis during 3-4 hours at 37 degree C, the concentration of biomass being 0.05 g of dry mycelium per 1 ml of hypertonic solution. Production of protoplasts was controlled by phase-contrast microscopy; the amount of the protoplast was determined by optical density at lambda=530 nm. The highest amount of the protoplasts was obtained during growth of the culture on a medium containing 0.5% glycine. The membrane fraction was isolated after lysis of the protoplasts in a hypotonic medium, treatment with DNase and RNase, and centrifugation at 22,000 g in the cold. Production of the membranes was controlled by electron microscopy. The yield of the membranes was 17-19% of the dry mycelium weight. They contained about 35% of lipids, 50% of protein, and 5% of RNA.
在含有10.3%蔗糖的高渗培养基中获得了吸水链霉菌原生质体。溶菌酶用作裂解剂，浓度为1毫克/毫升。细胞在37℃下进行3 - 4小时的裂解，生物量浓度为每1毫升高渗溶液含0.05克干菌丝体。原生质体的产生通过相差显微镜进行控制；原生质体的数量通过在λ = 530纳米处的光密度来确定。在含有0.5%甘氨酸的培养基上培养时，获得了最高数量的原生质体。在低渗培养基中裂解原生质体、用DNA酶和RNA酶处理并在冷条件下以22,000克离心后，分离出膜部分。膜的产生通过电子显微镜进行控制。膜的产量为干菌丝体重量的17 - 19%。它们含有约35%的脂质、50%的蛋白质和5%的RNA。
