[Isolation of the individual structural elements of bacteria of the genus Bordetella and a study of their properties. I. The formation of mureinoplasts and true protoplasts from B. pertussis].
B. pertussis suspension was tested by De Voe et al. method (1970) and its modification with the solutions of a definite ionic composition and a lysozyme. The best results were obtained by the following modification elaborated by the authors. The microbes were grown on the casein-carbon agar for 36 hours and were washed with chilled 0.5 M NaCl. The suspension was washed 4 times with the same solution and then the precipitate was suspended in saccharose solution (0.5 M). In 2 hours the saccharose was replaced by a solution of salts with lysozyme. After a 2-hour incubation at 35 degrees C the substance was centrifugated for 20 minutes and the precipitate suspended in the tris-buffer at pH 7.8. The following changes were observed: after the washing and incubation with saccharose there was seen a strong stretching and separation of the cell wall (CW) from the cytoplasmic membrane (CPM); cells without the CW were rarely revealed; 2) after the lysozyme treatment there were many cells of spherical shape (phasic-contrast microscopy) without any CW, limited by the CPM only. Morphologically they were no different from the true protoplasts of the Gram-positive bacteria. The chemical analysis also confirmed a possibility of obtaining the true protoplasts of the Gram-negative bacteria.
D001884:Bordetella / Q000302:isolation & purification*; D001886:Bordetella pertussis / Q000302:isolation & purification*; D002458:Cell Fractionation; D002473:Cell Wall / Q000648:ultrastructure*; D003584:Cytological Techniques; D006863:Hydrogen-Ion Concentration; D009113:Muramidase; D011523:Protoplasts / Q000648:ultrastructure*; D013053:Spectrophotometry; D013535:Suspensions; D014465:Ultrasonics
D013535:Suspensions; D009113:Muramidase
Vydelenie otdelnykh strukturnykh élementov bakteriĭ roda Bordetella i izuchenie ikh svoĭstv. Soobshchenie I. Obrazovanie mureinoplastov i istinnykh protoplastov iz B. pertussis