title: Visualization of endo-beta-N-acetylglucosaminidase, lysozyme, and lysostaphin after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate.
abstract: Bacteriolytic enzymes of different bond specificities, denatured by sodium dodecyl sulphate (SDS), were electrophoresed in polyacrylamide gels containing bacterial cells, then renatured after removal of SDS by diffusion. Enzyme activity was seen in sharp transparent bands resulting from bacteriolysis in the gels, while these sections containing bacterial cells appeared cloudy. Bacteriolytic enzymes including staphylococcal endo-beta-N-acetylglucosaminidase, lysozyme (N-acetylmuramidase), and lysostaphin (endopeptidase) were detected. The major bacteriolytic enzymes of Staphylococcus spp. were identified in gels after electrophoresis of crude enzyme preparations. This demonstrates the wide applicability of this method to the study of staphylococcal bacteriolytic enzymes. However, it should be noted that the method will fail to detect activities of bacteriolytic enzymes which are irreversibly inhibited by SDS.
authors.affiliation: Department of Microbiology, Hiroshima University School of Dentistry, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Sugai
authors.forename: H
authors.initials: H
authors.lastname: Komatsuzawa
authors.forename: Y
authors.initials: Y
authors.lastname: Miyake
authors.forename: H
authors.initials: H
authors.lastname: Suginaka
mesh_terms: D001433:Bacteriolysis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006026:Glycoside Hydrolases / Q000302:isolation & purification*; D008248:Lysostaphin / Q000302:isolation & purification*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000302:isolation & purification*; D011487:Protein Conformation; D011489:Protein Denaturation; D012967:Sodium Dodecyl Sulfate; D013194:Staining and Labeling
chemical_list: D012967:Sodium Dodecyl Sulfate; D006026:Glycoside Hydrolases; C026255:endo-N-acetylmuramidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008248:Lysostaphin