Characterization of Purified Shiga Toxin from Shigella dysenteriae 1

• Published in: MICROBIOLOGY and IMMUNOLOGY 1986, Vol.30(11), pp.1115-1127
• Main Authors: Yutsudo, Takashi, Honda, Takeshi, Miwatani, Toshio, Takeda, Yoshifumi
• Format: Article
• Language: English
• Subjects: Amino Acids - analysis; Animals; Bacterial Toxins - analysis; Bacterial Toxins - isolation & purification; Bacteriology; Biological and medical sciences; Cell Line; Cell Survival - drug effects; Chromatography, DEAE-Cellulose; Fundamental and applied biological sciences. Psychology; Haplorhini; Ileum - drug effects; Isoelectric Focusing; Lethal Dose 50; Mice; Microbiology; Molecular Weight; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains; Rabbits; Shiga Toxins; Shigella dysenteriae; Shigella dysenteriae - analysis; toxins
• ISSN: 0385-5600; 1348-0421
• DOI: 10.1111/j.1348-0421.1986.tb03041.x

Shiga toxin was purified from the culture supernatant of Shigella dysenteriae 1 by ammonium sulfate fractionation, DEAE-cellulose column chromatography and repeated chromatofocusing column chromatography. About 1.6mg of purified Shiga toxin was obtained from 15 liters of culture with a yield of about 27%. The molecular weight of purified Shiga toxin was estimated to be 62, 000. The toxin consisted of A and B subunits with molecular weights of about 30, 000 and 5, 000-6, 000, respectively. The isoelectric point of purified Shiga toxin was 7.0. Purified Shiga toxin showed the following biological activities: lethal toxicity to mice when injected intraperitoneally with an LD50 of 28ng per mouse; cytotoxicity to Vero cells, killing about 50% of the cells at 1pg and all of the cells at 10pg; and fluid accumulation in rabbit ileal loops at a concentration of more than 1μg.