Glucosamine substitution and muramidase susceptibility in Bacillus anthracis.

Cell walls of Bacillus anthracis were found to be resistant to lysozyme, and partially resistant to mutanolysin, a muramidase from Streptomyces globisporus. Following treatment with acetic anhydride, it was observed that the walls were highly susceptible to hydrolysis by lysozyme or mutanolysin. Analyses of cell walls, prior to and following derivatization with fluorodinitrobenzene, revealed that approximately 88% of the glucosamine residues and 34% of the muramic acid residues of the peptidoglycan contained unsubstituted amino groups, thereby providing an explanation for the resistance of the walls to lysozyme. The walls of B. anthracis were approximately 19% cross-linked, based on the findings that 81% of the diaminopimelic acid residues could be modified by fluorodinitrobenzene. Walls of B. thuringiensis 4040 and B. cereus ATCC 19637 also contained high percentages of unsubstituted amino sugars, and unless acetylated, were also relatively resistant to lysozyme and mutanolysin. When B. anthracis, B. cereus, or B. thuringiensis were grown in the presence of 100 micrograms/mL lysozyme, there was a decrease in the average number of cells per chain, but there was no decrease in growth rates, suggesting that the enzyme was acting at septa. It is unlikely that lysozyme and autolysins act synergistically in Bacillus, because azide anion, which activates autolysins, did not enhance the lytic action of lysozyme in B. anthracis, B. cereus, or B. thuringiensis.