Induction of L-phase variant from protoplast of Staphylococcus aureus

Protoplasts of Staphylococcus aureus 209P and Cowan 1 were induced by treatment with lysostaphin. These protoplasts were sensitive to detergent, a low concentration of sodium chloride and low temperature. Almost all protoplast cells spread on CLYS agar medium (casein hydrolysate, yeast extract, Na-lactate, and NaCl) formed typical L-form colonies. Horse serum (0.25%) and Mg2+ (100 mM) are essential factors for formation of the L-form colonies of 209P. In the case of Cowan 1, Mg2+ was not required. The active factor(s) in horse serum was heat-resistant and protein in nature.