title: [Production of protoplasts and membranes in Actinomyces hygroscopus].
abstract: 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.
mesh_terms: D000190:Actinomyces / Q000302:isolation & purification* / Q000648:ultrastructure; D002462:Cell Membrane / Q000648:ultrastructure; D003851:Deoxyribonucleases / Q000378:metabolism; D006982:Hypertonic Solutions; D009113:Muramidase / Q000378:metabolism; D011523:Protoplasts / Q000648:ultrastructure; D012260:Ribonucleases / Q000378:metabolism; D013395:Sucrose
chemical_list: D006982:Hypertonic Solutions; D013395:Sucrose; D003851:Deoxyribonucleases; D012260:Ribonucleases; D009113:Muramidase
vernacular_title: Poluchenie protoplastov i membran Actinomyces hygroscopus