Effect of penicillin G on release of peptidoglycan fragments by Neisseria gonorrhoeae: characterization of extracellular products.

Abstract

The effect of penicillin G (penG) on the turnover and release of peptidoglycan (PG) by Neisseria gonorrhoeae RD5 was investigated. We previously showed that exponentially growing gonococci (labeled in the glycan moiety with glucosamine and muramic acid and in the peptide with diaminopimelic acid) turn over ca. 35% of their PG per generation. In current studies, addition of penG accelerated the rate of PG hydrolysis by more than twofold and resulted in a corresponding increase in the amount of soluble PG fragments found in the medium. This increase of soluble PG was accounted for mainly by the release of nonreducing (anhydro-muramyl-containing) disaccharide peptide dimers (molecular weight, about 2,000) and trimers that were composed of subunits, linked not by peptide cross-linking bonds, but probably only by glycosidic bonds. The enhanced release of these products suggested that penG, directly or indirectly, stimulates the activity of a glycan-splitting, gonococcal PG:PG-6 muramyl transferase (transglycosylase). PG monomers that were released in the presence of penG were identical, both qualitatively and quantitatively, to the monomers released as a result of turnover in the absence of penG and consisted of anhydro-muramyl-containing disaccharide tripeptides and tetrapeptides. PenG-treated bacteria consistently released slightly less free disaccharide and free peptides than did control cultures, implying a penG-associated depression in the activity of the gonococcal N-acetylmuramyl-L-alanine amidase. In addition to stimulating the release of PG fragments, penG was also associated with greatly enhanced release from cells of glucosamine-containing non-PG macromolecule(s).