Isolation problems and structural organization of membrane units in Mycobacterium sp. smegmatis

Summary Membrane units from lysed spheroplasts induced by lysozyme or glycine from Mycobacterium spec. smegmatis were isolated in a biological active state by differential centrifugation and by density gradient technique. They were compared morphologically with membraneous fractions obtained from mycobacterial cells disintegrated under a high hydrostatic pressure.Higher homogeneity of membraneous structures isolated from spheroplasts was confirmed. Three types of membraneous structures could be distinguished. They include empty ghosts of spheroplasts, tubular structures containing cytoplasmic material and fragments of typical membraneous structures relatively free of contaminants. By studying protoplasts in the process of lysis it was determined that these structures correspond with cytoplasmic membranes and mesosomes.Differences between lysozome and glycine induced spheroplasts as a starting material for isolation of membraneous structures include the proportion of contamination by other cellular components, reasons of which are discussed.