Open AccessCharacterization of sodium dodecyl sulfate-stable Staphylococcus aureus bacteriolytic enzymes by polyacrylamide gel electrophoresis
Author(s) -
Motoyuki Sugai,
Tomoko Akiyama,
Hitoshi Komatsuzawa,
Yoichiro Miyake,
Hidekazu Suginaka
Publication year - 1990
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.172.11.6494-6498.1990
Subject(s) - micrococcus luteus , sodium dodecyl sulfate , staphylococcus aureus , gel electrophoresis , biochemistry , sodium , polyacrylamide gel electrophoresis , biology , chromatography , iodoacetic acid , membrane , enzyme , bacteria , chemistry , organic chemistry , genetics
Profiles of the bacteriolytic activities of Staphylococcus aureus culture supernatants, sodium dodecyl sulfate cell extracts, LiCl cell extracts, cell wall extracts, and cell membranes were analyzed in sodium dodecyl sulfate-polyacrylamide gels containing Micrococcus luteus or S. aureus. A total of 20 distinct bands of bacteriolytic activity could be detected in gels containing M. luteus, 8 of these bands were found in culture supernatants. The sodium dodecyl sulfate cell extracts, the LiCl cell extracts, and the cell membranes each contained 20 bands (P1 to P20), but no activity was found in cell wall extracts. Less bacteriolytic activity could be detected in gels containing S. aureus, although three bands were found in culture supernatants and LiCl extracts and cell membranes contained one major band, P13. Crude cell extracts showed five bacteriolytic bands of which the major bacteriolytic bands were distributed in an identical manner in all 10 strains of S. aureus studied. The effects of chemical and physical factors were determined, and it was shown that iodoacetic acid, Hg2+, and Cibacron Blue 3G-A reduced activity, and an optimum pH for enzyme detection was between 7 and 8. Preincubation at 100 degrees C for 30 min reduced the activity of P1 and P2 bands.