Studies on extracellular proteins from Staphylococcus aureus: VI. Production and purification of β-haemolysin in large scale

Abstract

Staphylococcal β-haemolysin was purified to a 40 000-fold increase in specific activity by CM-Sephadex C-25 chromatography, isoelectric focusing, and Biogel P-10 chromatography. Lyophilization was found to be the method of choice for storage of the unstable purified β-haemolysin. The high degree of purity obtained was investigated by acrylamide electrophoresis in continuous and discontinuous buffer systems and different gel concentrations. The microheterogeneity of the purified β-haemolysin with an isoelectric point ( pI ) of 9.4 was studied by electrophoresis and isoelectric focusing in acrylamide gels. The gels were sliced and the activity eluted. No further resolution in separable subcomponents was achieved. The main component ( pI 9.4 ± 0.1 ) of β-haemolysin was also devoid of all twelve enzymatic and toxic activities assayed for except sphingomyelinase, which gives further evidence for the identity of these two activities. Some of the biological properties, like the lethal effect and cytotoxicity, have been investigated (trö and ö, Biochim. Biophys. Acta), (1971).