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Purification and Properties of Staphylococcal -Hemolysin I. Production of -Hemolysin¹

^a Department of Microbiology, University of Illinois at the Medical Center, Chicago, Illinois 60680

ABSTRACT

Concentrated preparations of staphylococcal -hemolysin were obtained by growing selected hemolytic colonies from the 146P strain of Staphylococcus aureus on dialysis membranes laid over Brain Liver Heart agar plates at 37 C for 20 hr under 10% CO₂ and harvesting the growth from five such membranes in 1.0 ml of deionized distilled water. Incubation in a humid environment facilitated this harvesting procedure. Incubation longer than 40 hr or incubation under CO₂ higher than 10 to 20% gave lower yields of -lysin. Addition of a sugar, fermented by the organisms, resulted in lower yields of -hemolysin. Agar, although separated from the growing cells by the dialysis membrane, did potentiate -hemolysin production. Addition of 0.1% agar to the inoculum further enhanced this potentiation. -Hemolysin produced in broth or semisolid cultures was excessively diluted with the media. Dialysis membranes prevented this dilution and thus yielded concentrated preparations of -hemolysin.

¹ Taken in part from a thesis submitted by R. Murphy to the Graduate College of the University of Illinois in partial fulfillment of the requirements for the M.S. degree.