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Schindler & Schuhardt (1964) reported on the isolation of an organism of the genus Staphylococcus that produced a lytic agent, lysostaphin, active against all of 59 strains of staphylococci tested. The specificity for organisms of the genus Staphylococcus separated its mode of action from that of lysozyme. The authors also described an assay for determining the quantity of enzyme based upon the decrease in turbidity of a standard staphylococcal strain. Since the rate of lysis of a staphylococcal strain was a function of the concentration of lysostaphin present, it was possible to deviate from the standard assay micro-organism in these experimental procedures. Schindler & Schuhardt (1965) purified and described some of the properties of lysostaphin and reported that it was a basic protein essentially devoid of cystine and cysteine and contained large quantities of aspartic and glutamic acid compared to the total amino acid content. The lytic protein had a sedimentation coefficient (S2,,,J of 2.355' and an isoelectric point between pH 10.4-11.4. The rate of lysis of Staphylococcus aureus by lysostaphin was dependent upon the concentration of NaCl present in the medium. This phenomenon paralleled the salt requirement of the muramidase lysozyme (Salton, 1957). By increasing the concentration of NaCl in the suspending medium from zero to 0.15 M, an increased rate of lysis of staphylococci was observed. Concentrations of salt above physiological values introduced parameters which affected both the bacterial cells and the stability of lysostaphin; thus such levels of salts were not evaluated. This investigation was undertaken to determine in what manner NaCl entered into the dynamics of the lysostaphin-substrate interaction. METHODS Micro-organism. Staphylococcus aureus, strain sv, the mouse-virulent variant of the Smith strain (Hunt & Moses, 1958) was grown in Trypticase soy broth at 37" under conditions of aeration and agitation to give a maximum yield of cocci/ml. 13-2

The Role of NaCl in the Lysis of Staphylococcus aureus by Lysostaphin