Purification and Some Properties of Progenitor Toxins of Clostridium botulinum Type B

Abstract

Purification of progenitor toxin of Clostridium botulinum type B strain Okra was undertaken by sequential steps of acid precipitation, extraction, ammonium sulfate precipitation, ribonuclease digestion, acid precipitation, protamine treatment, sulphopropyl-Sephadex chromatography, and Sephadex G-200 gel filtration. Two different molecular-sized toxins, named large (L) and medium (M) toxins, were obtained. L toxin was centrifugally homogeneous but electrophoretically heterogeneous. It contained 2.5 x 108 to 3.0 x 108 mean lethal doses per mg of nitrogen, and its sedimentation constant was 16S. M toxin was centrif'ugally and electrophoretically homogeneous. It contained 5.5 x 108 to 6.0 x 108 mean lethal doses per mg of nitrogen, and its sedimentation constant was 12S. The presence of both L and M toxins in spent culture was demonstrated. It seems justified, therefore, to call both progenitor toxins. Both consisted of toxic and nontoxic components. The toxic components of L and M toxins appeared to be identical with each other. The nontoxic component of L toxin was 12S and

Antigenic structure of Clostridium botulinum type B neurotoxin and its interaction with gangliosides, cerebroside, and free fatty acids

“…Type B progenitor toxin was prepared by a previously published method (16). The neurotoxin was purified by DEAE-Sephadex A-50 (Pharmacia, Uppsala, Sweden) chromatography and converted to toxoid by dialysis against 0.1 M phosphate buffer, pH 7.0, containing 0.4% Formalin (14,17). The purified neurotoxin proved to be unnicked by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (18).…”

Botulogenic Properties of Vegetables With Special Reference to the Molecular Size of the Toxin in Them

“…The existence of heterogenously molecular-sized purified type . A and B toxins, 12, 16 and 19s for type A , and 1 2 and 1 6 s for type B (Kozaki et al 1974) have been demonstrated. Type E and F toxins were reported t o be uniform in molecular size, at 12 and lOS, respectively (Kitamura et a / .…”

“…Type B strains produce both 12 and 16s toxins in culture (Kozaki et al 1974). Schantz and Spero (1967) reported that type A strains produce 1 9 s toxin only in culture, whereas Sugii and Sakaguchi (1975) demonstrated that they produce only 12s toxin in culture.…”

Comparison of progenitor toxins of nonproteolytic with those of proteolytic Clostridium botulinum Type B

“…Nonproteolytic strains of types B, E, and F are known to produce toxins that are activated upon trypsinization (2,3,5). The progenitor toxin of a proteolytic type B strain Okra was obtained in two different molecular forms, 16S (L toxin) and 12S (M toxin) (9). Both L and M toxins consist of neurotoxic and nontoxic components.…”

“…The neurotoxic component of either toxin has a molecular weight of 170,000 (7S) (1,8). The nontoxic component of L toxin is about 12S and contains hemagglutinin activity, whereas that of M toxin is 7S and contains no hemagglutinin activity (9).…”