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PURIFICATION OF TYPE B STAPHYLOCOCCAL ENTEROTOXIN¹

^a Departments of Bacteriology and Biochemistry, University of Wisconsin, Madison, Wisconsin

ABSTRACT

FREA, JAMES I. (University of Wisconsin, Madison), ELIZABETH MCCOY, AND F. M. STRONG. Purification of type B staphylococcal enterotoxin. J. Bacteriol. 86:1308–1313. 1963.—A procedure for the production and isolation of type B staphylococcal enterotoxin in yields up to 4 mg per liter of toxic culture is presented. Staphylococcus aureus S6 was grown in shake flask cultures at 37 C for 24 hr in a medium consisting of 2% Hy-Case S.F., 0.3% yeast extract, calcium pantothenate, nicotinic acid, and thiamine. Cells were removed by centrifugation and the supernatant fluid was concentrated by dialysis against polyethylene glycol. Enterotoxin was isolated from the concentrate by ethanol precipitation, gel filtration on a G-100 Sephadex column, and Sephadex-column electrophoresis. The purified toxin was identical to type B reference enterotoxin on both disc electrophoresis and Ouchterlony gel diffusion. It contained no carbohydrates, lipids, or nucleic acids, gave positive tests for protein; and contained 11.6% N. The preparation showed no coagulase, alpha-hemolysin, or deoxyribonuclease activity. Emesis in monkeys resulted when the experimental toxin was injected at the rate of 0.26 µg of N per kg of body weight.

² National Science Foundation predoctoral fellow. Present address: First USA Medical Laboratory, 90 Church Street, New York, N.Y.

¹ Published with the approval of the Director of the Wisconsin Agricultural Experiment Station.