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J Bacteriol. 1964 February; 87(2): 401-407
Copyright © 1964 by the American Society for Microbiology. All Rights Reserved.

COMPARATIVE GEL FILTRATION OF TOXIN PRECURSOR AND TRYPSIN-ACTIVATED TOXIN OF CLOSTRIDIUM BOTULINUM TYPE E

Department of Food Control, National Institute of Health, Tokyo, Japan

ABSTRACT

SAKAGUCHI, GENJI (National Institute of Health, Tokyo, Japan), SUMIKO SAKAGUCHI, AND NOBUKO IMAI. Comparative gel filtration of toxin grecursor and trypsin-activated toxin of Clostridium botulinum type E. J. Bacteriol. 87:401–407. 1964.—Precursor of type E botulinus toxin was highly purified from bacterial cells by extraction, ammonium sulfate precipitation, ribonuclease digestion, and chromatography on CM-Sephadex. The sample free from ribonucleic acid had a toxicity of 5.1 x 10⁵LD₅₀ per mg of nitrogen before activation and 8.3 x 10⁷LD₅₀ per mg of nitrogen after activation. This precursor and its activated product were subjected to gel filtration on a column of Sephadex G-200. No evidence for smaller fractions was obtained. Both precursor and trypsin-activated toxin were eluted in the void volume with 0.05 or 1 M acetate buffer (pH 6.0) or with 0.05 or 0.5 M phosphate buffer (pH 7.5). Intact trypsin and its degradation products were separated from toxin. The toxins eluted with the acetate buffers had potencies of 1.2 x 10⁸ and 1.3 x 10⁸LD₅₀ per mg of N, while those eluted with the phosphate buffers showed lower toxicities. Possible mechanisms involved in the activation process are discussed.