Abstract
Sporogenic and asporogenicBacillus megaterium strains, as well asBacillus cereus degraded the murein component of the cell wall labelled with14C-diaminopimelic acid to TCA-soluble compounds during growth. The rate of murein turnover was about 15% during one generation in all three cases. The addition of chloramphenicol instantaneously markedly decreased the degradation rate, whereas in the presence of penicillin the degradation proceeded at the beginning at a rate comparable with that in the control and decreased only after a certain time interval. The cell wall degradation was considerably or completely stopped during the stationary phase of growth. In sporogenic strains ofBacillus megaterium andBacillus cereus the release of mature spores was associated with a new wave of the wall degradation, during which the wall of the sporangial cell was completely digested to TCA-soluble fragments. Free spores contained practically no mucopeptide component (cortex or spore wall) originating from the wall of the vegetative cell. A possible existence of a stable fraction of the cell wall not subject to turnover was investigated by measuring the3H/14C ratio in cells labelled simultaneously with3H (or14C)-diaminopimelic acid and14C (or3H)-leucine. The ratio changed during five generations, remaining constant later. This indicates that a certain portion of murein could be stable. The murein degradation during growth was not associated with secretion or release of a significant quantity of autolytic enzymes into the medium. The wall was apparently attacked from the inside. On the other hand, the release of the spore was accompanied by an increasing autolytic activity in the medium. This latter activity reached values corresponding to 3–8 μg lysozyme/ml.
Similar content being viewed by others
References
Chaloupka, J.:Synthesis and degradation of surface structures by growing and non-growing Bacillus megaterium. Fol. microbiol. 12: 264, 1967.
Chaloupka, J.:Surface structures in the biology of the bacterial cell. Rozpravy ČSAV 81: 1, 1971.
Chaloupka, J., Křečková, P.:Regulation of the formation of protease in Bacillus megaterium. I. The influence of amino acids on the enzyme formation. Fol. microbiol. 11: 82, 1966.
Chaloupka, J., Křečková, P., Říhová, L.:The mucopeptide turnover in the cell walls of growing cultures of Bacillus megaterium KM. Experientia 18: 362, 1962.
Chaloupka, J., Říhová, L., Křečková, P.:Degradation and turnover of bacterial cell wall mucopeptides in growing bacteria. Fol. microbiol. 9: 9, 1964.
Chatterjee, A. N., Mirelman, D., Singler, H. J., Park, J. T.:Properties of a novel pleiotropic bacteriophage-resistant mutant of Staphylococcus aureus H. J. Bacteriol. 100: 846, 1969.
Conover, H. J., Thompson, J. S., Shockman, G. D.:Autolytic enzyme of Streptococcus faecalis: release of soluble enzyme from cell walls. Biochem. biophys. Res. Commun. 23: 713, 1966.
Ellwood, D. C., Tempest, D. W.:Control of teichoic acid and teichuronic acid biosynthesis in chemostat cultures of Bacillus subtilis var. niger. Biochem J. 111: 1, 1969.
Fan, D. P.:Autolysin (s) of Bacillus subtilis as dechaining enzyme. J. Bacteriol. 103: 494, 1970.
Higgins, M. L., Pooley, H. M. Shockman, G. D.:Site of initiation of cellular autolysis in Streptococcus faecalis as seen by electron microscopy. J. Bacteriol. 103: 504, 1970.
Hisatsune, H., De Courcy, S. J. Jr., Mudd, S.:Studies on the carbohydrate—peptide fraction of the centrifuge supernatants of Staphylococcus aureus cultures: The immunologically active cell wall peptide polymer of Staphylococcus aureus. Biochemistry 6: 595, 1967.
Lowry, H. O., Rosenbrough, N. J., Farr, A. L., Randall, R. J.:Protein measurement with the folin phenol reagent. J. biol. Chem. 193: 265, 1951.
Mauck, J., Glaser, L.:Turnover of the cell wall of Bacillus subtilis during logarithmic growth. Biochem. biophys. Res. Commun. 39: 699, 1970.
McQuillen, K.:Bacterial protoplasts: growth and division of protoplasts of Bacillus megaterium. Biochem. biophys. Acta 18: 485, 1955.
Mitchell, P., Moyle, J.:Autolytic release and osmotic properties of “protoplasts” from Staphylococcus aureus. J. gen. Microb. 16: 184, 1957.
Park, J. T., Hancock, R.:A fractionation procedure for studies of the synthesis of cell wall mucopeptide and of other polymers in cells of Staphylococcus aureus. J. gen. Microb. 22: 249, 1960.
Richmond, M. H.:Formation of a lytic enzyme by a strain of Bacillus subtilis. Biochem. biophys. Acta 33: 78, 1959.
Rogers, H. J.:The outer layers of bacteria: The biosynthesis of structure. In “Function and structure in micro-organisms”. Cambridge Univ. Press, p. 186, 1965.
Rogers, H. J.:The structure and biosynthesis of the walls of gram positive bacteria. Fol. microbiol. 12: 191, 1967.
Schwarz, U., Weidel, W.:Zum Wirkungsmechanisms von Penicillin. II. Nachweis eines Penicillin-induzierten hydrolytischen Abbaus von Murein and Mureinvorstufen in E. coli B. Z. Naturforsch. 20 B/153, 1965.
Shockman, G. D., Pooley, H. M., Thompson, J. S.:Autolytic enzyme system of Streptococcus faecalis. III. Localization of autolysin at the sites of the cell wall synthesis. J. Bacteriol. 94: 1525, 1967.
Shockman, G. D., Thompson, J. S., Conover, M. J.:The relationship of autolysin to lysozyme sensitivity of Streptococcus faecalis 9790. In L. B. Guze: “Microbiol. protoplasts, spheroplasts and L-forms”. William and Wilkins Comp., Baltimore p. 248, 1968.
Weidel, W., Pelzer, H.:Bagshaped mocromolecules—a new outlook on bacterial cell walls. Adv. Enzymology 26: 193, 1964.
Young, F. E.:Autolytic enzyme associated with cell walls of Bacillus subtilis. J. biol. Chem. 241: 3462, 1966.
Urbà, R. C.:Protein breakdown in Bacillus cereus. Biochem. J. 71: 513, 1959.
Vinter, V.:Spores of microorganisms. VIII. The synthesis of specific calcium and cystine containing structures in sporulating cells of bacilli. Fol. microbiol. 5: 217, 1960.
Vinter, V.:Spores of microorganisms. XII. Nonparticipation of the preexisting sporangial cell wall in the formation of spore envelopes and gradual synthesis of DAP-containing structures during sporogenesis of bacilli. Fol. microbiol. 8: 147, 1963.
Author information
Authors and Affiliations
Additional information
The results published here were presented at the 2nd Harden Conference “Cell walls and cell membranes”, Wye, Kent, England in September 1970.
Rights and permissions
About this article
Cite this article
Chaloupka, J., Křečková, P. Turnover of mucopeptide during the life cycle ofBacillus megaterium . Folia Microbiol 16, 372–382 (1971). https://doi.org/10.1007/BF02875757
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02875757