Bacteriolytic enzymes from Staphylococcus aureus. Purification of an endo-beta-N-acetylglucosaminidase.
On cultivation of Staphylococcus aureus in a complex liquid medium, bacteriolytic activity is found extracellularly. The maximal amount was found at the end of the exponential growth phase in batch culture, but in continuous culture run under similar conditions the yield was doubled. Isoelectric focusing of dialysed crude culture supernatants showed that the bacteriolytic activity of all four strains studied (M18, 524, Wood 46 and Duncan) was heterogeneous. The most alkaline peak of activity (isoelectric point 9.5+/-0.1) was assayed against Micrococcus lysodeikticus turbidimetrically. This bacteriolytic activity was purified more than 70-fold after continuous dialysis by adsorption on CM-Sephadex, precipitation with ethanol, heat purification, isoelectric focusing and Sephadex G-100 chromatography. The purified enzyme (isoelectric point 9.6+/-0.1) was found to give a single band on polyacrylamide-gel and cellulose acetate electrophoresis and was devoid of all 14 staphylococcal enzymes and toxins assayed for. The molecular weight is 70000+/-5000 as estimated by Sephadex G-100 and G-200 chromatography. The marked instability of the partially and highly purified enzyme was investigated. The mode of action and some properties of this enzyme are given in the following papers (Wadström & Hisatsune, 1970; Wadström, 1970). These results indicate that this extracellular enzyme which is produced by several strains of S. aureus is not a ;lysozyme' (endo-beta-N-acetylmuramidase) as previously suggested, but an endo-beta-N-acetylglucosaminidase.
D000327:Adsorption; D001431:Bacteriological Techniques; D001433:Bacteriolysis; D001681:Biological Assay; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D003720:Densitometry; D003956:Dialysis; D004586:Electrophoresis; D000431:Ethanol; D006026:Glycoside Hydrolases / Q000302:isolation & purification* / Q000494:pharmacology; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007525:Isoelectric Focusing; D008837:Micrococcus / Q000187:drug effects; D008970:Molecular Weight; D013210:Staphylococcus / Q000201:enzymology*
D003470:Culture Media; D000431:Ethanol; D006026:Glycoside Hydrolases