Extracellular cell wall lytic enzyme from Staphylococcus aureus: purification and partial characterization.
An autolysin obtained from culture fluid of Staphylococcus aureus strain 8507 was purified 3,000-fold. One milligram of this preparation (S-5DL) will solubilize 12 mg of cell wall in 1 hr. The major activity is N-acetylmuramyl-l-alanine amidase. Recovery of lytic activity in the purified preparation was repeatably only 20% of the starting level. This suggests that other cell wall lytic enzymes may be present in the starting material. The S-5DL enzyme has been compared to freeze-thaw extracted enzyme (AFZ). Both enzymes precipitate in 0.01 m KPO(4) (pH 6.0) and dissolve in 0.1 to 0.7 m NaCl. Fifty per cent of the AFZ activity and 66% of the S-5DL activity bind rapidly to cell walls of S. aureus at 0 C in the presence of magnesium ion. None of the AFZ activity and 66% of the S-5DL activity bind to cell walls at 0 C in the absence of magnesium ion. The cell walls of nine different strains of S. aureus were compared for level of native autolysin activity. These same walls after inactivation of the native autolysin were tested for susceptibility to the S-5DL enzyme.
D000581:Amidohydrolases / Q000378:metabolism; D001329:Autolysis; D001433:Bacteriolysis; D002473:Cell Wall / Q000187:drug effects / Q000201:enzymology*; D011232:Chemical Precipitation; D002855:Chromatography, Thin Layer; D004798:Enzymes / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D005557:Formaldehyde; D005612:Freeze Drying; D006863:Hydrogen-Ion Concentration; D007477:Ions; D008274:Magnesium / Q000494:pharmacology; D010759:Phosphorus Isotopes; D000644:Quaternary Ammonium Compounds; D012995:Solubility; D013045:Species Specificity; D013210:Staphylococcus / Q000201:enzymology*; D013431:Sulfates
D004798:Enzymes; D007477:Ions; D010759:Phosphorus Isotopes; D000644:Quaternary Ammonium Compounds; D013431:Sulfates; D005557:Formaldehyde; D000581:Amidohydrolases; D008274:Magnesium