The bactericidal action of beta-lactam antibiotics on an autolysin-deficient strain of Bacillus subtilis

An autolysin-deficient mutant of Bacillus subtilis was completely tolerant to 5 h incubation with 50-100 micrograms cycloserine ml-1 whereas the wild-type was rapidly lysed and killed by 12 micrograms ml-1. Lysis also did not occur when low concentrations of beta-lactams were added to exponentially growing cultures of the mutant, but over 90% of the bacteria were killed within 90-120 min. Protein, lipid and peptidoglycan synthesis as well as growth were inhibited after about 60 min. At this time, but not earlier, small amounts of these three cell components appeared in culture supernatants. Earlier, at about 20-30 min, the intracellular pools of amino acids started to decline rapidly and there was a temporary apparent increase in the rate of lipid synthesis. Neither of the latter phenomena occurred with cycloserine, with which protein and lipid synthesis declined only slowly and the rate of peptidoglycan synthesis was 80% inhibited within 30 min. Only occasional cells with damaged walls were seen 30-90 min after addition of either beta-lactams or cycloserine to the cultures. It thus seems unlikely that wall hydrolysis or penetration by residual autolysins in the mutant are responsible for mass cell death caused by the beta-lactams.