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J. Dairy Sci. 86:96-104
© American Dairy Science Association, 2003.

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

O. J. Kang*, S. Laberge{dagger} and R. E. Simard*

* Faculté des Sciences de l’Agriculture et de l’Alimentation, Centre de Recherche en Sciences et Technologie du Lait, Université Laval, Québec, Canada, G1K 7P4
{dagger} Agriculture et Agroalimentaire Canada, Centre de Recherche et de Développement sur les Sols et les Grandes Cultures, Ste-Foy, Québec, Canada, G1V 2J3

Corresponding author:
S. Laberge; e-mail:
laberges{at}agr.gc.ca.

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.

Key Words: peptidoglycan hydrolase • electron and fluorescence microscopy • Lactobacillus

Abbreviation key: DIC = differential interference contrast, FITC = fluorescein isothiocyanate, FLM = fluorescence microscopy, GAM = goat anti-mouse, LRW = London resin white, OD = optical density, TEM = transmission electron microscopy






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