PREPARATION OF CELL-FREE STAPHYLOCOCCAL PENICILLINASE CONCENTRATE

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Abstract
The preparation of staphylococcal intracellular penicillinase as a cell-free concentrate is described and the optimal conditions for its action on penicillin indicated. Cells were disrupted by sonic vibrations and the debris removed by centrifugation at 17000 × g. Cell-free concentrates were prepared from the supernatants and were found to have a higher activity per mg nitrogen than the bacterial suspensions from which they were prepared. Centrifugation at 35000 × g resulted in a partial and as yet unaccountable loss of activity. The enzyme was optimally active at pH 6.0–6.5 and 30 °C. The possible application of cell-free staphylococcal penicillinase to enzymatic and immunological work is discussed.