Peptidoglycan synthesis in Bacillus licheniformis. The inhibition of cross-linking by benzylpenicillin and cephaloridine in vivo accompanied by the formation of soluble peptidoglycan.
The synthesis of peptidoglycan by an autolysin-deficient beta-lactamase-negative mutant of Bacillus licheniformis was studied in vivo in the absence of protein synthesis. Benzylpenicillin and cephaloridine inhibited the formation of cross-bridges between newly synthesized peptidoglycan and the pre-existing cell wall. This inhibition, detected by measurement of the incorporation of N-acetyl[14C]glucosamine into the glycan fraction of the cell wall, was reversed by treatment with beta-lactamase and washing. Inhibition of D-alanine carboxypeptidase by benzylpenicillin was not reversed under similar conditions. Cells in which the initial penicillin inhibition of transpeptidation had been reversed showed an increased sensitivity to a subsequent addition of the antibiotic. Chemical analysis of peptidoglycan synthesized after reversal of penicillin inhibition revealed the presence of excess of alanine resulting from the continued inhibition of D-alanine carboxypeptidase. When the cell walls were digested to yield muropeptides so that the degree of cross-linking could be measured, the product after reversal of penicillin inhibition contained fewer cross-links than did the control preparation. Cultures treated with benzylpenicillin and cephaloridine continued to synthesize uncross-linked soluble peptidoglycan, which accumulated in the medium. This soluble material was all newly synthesized peptidoglycan and did not result from autolysis of the bacteria. The average chain lengths of the glycan synthesized in vivo and released as soluble peptidoglycan in the presence of both benzylpenicillin and cephaloridine were similar to those found previously in this organism.
D000117:Acetylglucosamine / Q000378:metabolism; D001407:Bacillus / Q000187:drug effects / Q000378:metabolism*; D001665:Binding Sites; D002268:Carboxypeptidases / Q000037:antagonists & inhibitors; D002473:Cell Wall / Q000378:metabolism; D002509:Cephaloridine / Q000494:pharmacology*; D002854:Chromatography, Paper; D009154:Mutation; D010400:Penicillin G / Q000494:pharmacology*; D010406:Penicillins / Q000037:antagonists & inhibitors; D010457:Peptidoglycan / Q000096:biosynthesis*
D010406:Penicillins; D010457:Peptidoglycan; D002268:Carboxypeptidases; D002509:Cephaloridine; D010400:Penicillin G; D000117:Acetylglucosamine