細菌毒素の本態に關する生化學的研究 : ヂフテリア毒素の純化とその組成

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Although many reports have described highly purified diphtherial toxin, there is no information about the immunologically and electrophoretically homogeneous preparations. We have obtained the culture filtrates (Lf 90∿100u/cc), consisting of beef extract and beef digested with papain, inoculated with Corynebacterium diphtheriae Park Williams No. 8 TORONTO. And have obtained 0.4g of protein from each 10 litre of the filtrates, by repeated fractionating precipitation with between 50∿60% saturation of ammonium sulfate, further precipitation with acetic acid at pH 4.1,resolution with weak alkali at pH 11,dialyzing against distilled water, and finally drying in cold vacuum. The preparation, a light brownish powder, contains approximately 370,000 Lf units per g, and 0.00044mg nitrogen per Lf. And 0.001γ<M. L. D.<0.01γ (for guinea pigs) was found. It behaves as a single protein in the Tiseliu's electrophoresis cell and RAMON's quantitative flocculation test and antigen-antibody reaction in agar-gel. Chemical analyses of the preparation showed that it is an iron-containing beta-globulin. The pure toxin and formalin-toxoid were compared with by protein color reactions, ultraviolet ray absorption test, and two dimendional paper chromatography with hydrochloric acid hydrolysates. Trytophane, aspartic acid, cystine, α-ε-diaminipimelic acid glutamic acid, serine, glycine, lysine, threonine, alanine, histidine, oxyproline, α or γ-aminobutyric acid, methionine, valine, proline, phenylalanine, leucine, isoleucine, tyrosine, and arginine were found in the pure toxin. The essential difference between the toxin and formalin-toxoid was found on the presence or absence of tyrosine residues in the molecules. Some possible structure concerning the molecules of the toxin and toxoid was suggested.
北海道大学の論文

細菌毒素の本態に關する生化學的研究 : ヂフテリア毒素の純化とその組成

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