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.
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Adámek, L., Mišoň, P., Mohelská, H., Trnka, L.: Ultrastructural organization of spheroplasts induced in Mycobacterium sp. smegmatis by lysozyme or glycine. Arch. Mikrobiol. (in press).
Akamatsu, Y., Ono, Y., Nojima, S.: Phospholipid patterns in subcellular fractions of Mycobacterium phlei. J. Biochem. (Tokyo) 59, 176–182 (1966).
Brieger, E. M., Glauert, A. M., Allen, J. M.: Cytoplasmic structures in Mycobacterium leprae. Exptl. Cell Res. 18, 418–421 (1959).
Chapman, G. B., Hanks, J. H., Wallace, J. H.: An electron microscope study of the disposition and fine structures of M. lepraemurium in mouse spleen. J. Bact. 77, 205–211 (1959).
Difco manual of dehydrated culture media and reagents for microbiological and clinical laboratory procedures, pp. 105. Detroit: Difco Laboratories, Inc. 1953.
Drews, G.: Elektronenmikroskopische Untersuchungen an Mycobacterium phlei. (Struktur und Bildung der metachromatischen Granula). Arch. Mikrobiol. 35, 53–62 (1960).
Granboulan, P., Leduc, E. H.: Ultrastructural cytochemistry of Bacillus subtilis. J. Ultrastruct. Res. 20, 111–126 (1967).
Fitz-James, P. C.: Participation of the cytoplasmic membrane in the growth and spore formation of bacilli. J. biophys. biochem. Cytol. 8, 507–528 (1960).
Imaeda, T., Ogura, M.: Formation of intracytoplasmic membrane system of Mycobacteria related to cell division. J. Bact. 85, 150–163 (1963).
—, Convit, J.: Electron microscope study of Mycobacterium leprae and its environment in a vesicular leprous lesion. J. Bact. 83, 43–52 (1962).
Iterson, W., van: Bacterial cytoplasm. Symposium on the fine structure and replication of bacteria and their parts. Bact. Rev. 29, 299–325 (1965).
Kleinzeller, A. (ed.): Manometric methods and their use in biology and biochemistry, pp. 365. Prague: SZN 1964.
Koike, M., Takeya, K.: Fine structures of intracytoplasmic organelles of mycobacteria. J. biophys. biochem. Cytol. 9, 597–608 (1961).
Pollard, E. C., Weller, P. K.: The effect of hydrostatic pressure on the synthetic processes in bacteria. Biochim. biophys. Acta (Amst.) 117, 573–580 (1966).
Reynolds, E. S.: The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol. 17, 208–212 (1963).
Salton, M. R. J.: The bacterial cell wall, pp. 215. Amsterdam, London, New York: Elsevier Publ. Comp. 1964.
Sauton, M. B.: Sur la nutrition minérale du bacille tuberculeux. Compt. rend. 155, 860–862 (1912).
Takeya, K., Koike, M., Yuda, Y., Mori, R., Nakashima, N., Tokunaga, T., Hagiwara, Y.: Electron microscopic studies on intracellular structures of Mycobacterium tuberculosis. Ann. Rep. Japan. Soc. Tuberc. 4, 1–5 (1959).
Trnka, L., Staflova, S., Nejedlý, J., Volný, B.: A procedure for disintegration of mycobacteria in cooled pressure chamber. Amer. Rev. resp. Dis. 93, 291–295 (1966).
—, Smith, D. W.: Influence of cell disruption methods on polyuridylic acid dependent polyphenylalanine synthesis by isolated ribosomes of mycobacteria. Experientia (Basel) 24, 1109–1110 (1968).
Weibull, C., Bergström, L.: The chemical nature of the cytoplasmic membrane and cell wall of Bacillus megaterium, strain M. Biochim. biophys. Acta (Amst.) 30, 340–351 (1958).
Yamaguchi, J.: Electron microscopic studies of tubercle bacilli. VI. Isolation and fine structure of cellular components of mycobacteria. Sci. Rep. Res. Inst. Tohoku Univ. Ser. C (Medicine). The Repts. Res. Inst. Tbc & Leprosy. 9, 125–146 (1960).
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Mišoň, P., Trnka, L., Mohelská, H. et al. Isolation problems and structural organization of membrane units in Mycobacterium sp. smegmatis . Archiv. Mikrobiol. 69, 216–226 (1969). https://doi.org/10.1007/BF00408974
- Hydrostatic Pressure
- Membraneous Fraction