Detection and Localization of a Peptidoglycan Hydrolase in Lactobacillus delbrueckiisubsp. bulgaricus

Peptidoglycan hydrolase activities in Lactobacillus delbrueckii subsp. bulgaricus were detected by analysis of bacterial extracts on denaturing polyacrylamide gel electrophoresis containing lyophilized Micrococcus lysodeikticus cells as substrate. A hydrolase with an estimated molecular mass of 80 kDa was found to cross-react on Western blot with monoclonal antibodies raised against muramidase-2 of Enterococcus hirae. These antibodies were also used to demonstrate that the method of cell sample preparation affected protein detection. Slot and Western blots indicate that the peptidoglycan hydrolase from L. bulgaricus is bound to the cell wall. Immuno-labeling followed by optical and electron microscopic observations suggest that this hydrolase is intracellular and restricted mainly to the space between the membrane and the cell wall.