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Pneumolysin was found to be produced by 112 of 113 clinical isolates of Streptococcus pneumoniae and to be an intracellular hemolysin. A 10-liter-scale fermentor production and purification procedure was developed for this hemolysin. The culture was concentrated by filtration 10 times before centrifugation. The cellular content was purified by ion-exchange chromatography, covalent thiopropyl gel chromatography, and gel filtration. One batch operation resulted in 6 mg of highly purified pneumolysin, with a yield of 66% and a specific activity of 1,400,000 hemolytic units per mg. The pneumolysin had a molecular weight of 53,000 and an isoelectric point of 5.2. The purification method developed will be of value in future studies on this hemolysin.
Kanclerski et al. (1987) studied this question.