Detection and localization of a peptidoglycan hydrolase in Lactobacillus delbrueckii subsp. 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.
Kang
O J
OJ
Faculty des Sciences de l'Agriculture et de l'Alimentation, Centre de Richerche en Sciences et Technologie du Lait, Université Laval, Québec, Canada, G1K 7P4.
Laberge
S
S
Simard
R E
RE
D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D002473:Cell Wall / Q000201:enzymology; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D016983:Enterococcus / Q000201:enzymology; D007150:Immunohistochemistry; D007778:Lactobacillus / Q000201:enzymology* / Q000254:growth & development / Q000648:ultrastructure; D008837:Micrococcus / Q000254:growth & development / Q000378:metabolism; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D009113:Muramidase / Q000276:immunology; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D009113:Muramidase; D009238:N-Acetylmuramoyl-L-alanine Amidase