PURIFICATION OF STAPHYLOCOCCAL COAGULASE

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Abstract
Nonhemolytic mutants of 2 coagulase producing strains of S. aureus were selected for production of purified coagulase. The enzyme was concentrated by acid precipitation from broth culture supernatant at pH 3.8 followed by 2 cycles of precipitation with ethanol (final concentration 70%). Further purification was achieved by starch column electrophoresis and anion exchange cellulose chromatography. Electrophoretically purified coagulase, which was not completely homogeneous by electrophoresis, contained one minimal clotting dose per 0.05 mg N. Overall purification factor was 387.5, yield 20.8%. Final product was antigenic for rabbits, which produced coagulase inhibiting antibodies.