Purification and Characterization of Streptococcal Erythrogenic Toxin Type A Produced by Streptococcus pyogenes Strain NY-5 Cultured in the Synthetic Medium NCTC-135. Comparison with the Dialyzed Medium (TP Medium)-Derived Toxin

Streptococcal erythrogenic toxin type A (ET-A) was purified from culture filtrate of Streptococcus pyogenes strain NY-5 grown in a chemically defined synthetic medium NCTC-135. We succeeded in simplifying the purification procedure, and obtained a highly purified preparation of ET-A. The purificatio...

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Bibliographic Details
Published in:MICROBIOLOGY and IMMUNOLOGY 1989, Vol.33(3), pp.183-194
Main Authors: Kamezawa, Yukio, Nakahara, Takako
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino Acids - analysis
Bacterial Proteins
Bacteriology
Biological and medical sciences
Chemical Phenomena
Chemistry
Chromatography, Gel
Chromatography, High Pressure Liquid
Culture Media - metabolism
Electrophoresis, Polyacrylamide Gel
Exotoxins - genetics
Exotoxins - isolation & purification
Exotoxins - toxicity
Fundamental and applied biological sciences. Psychology
Immunodiffusion
Membrane Proteins
Microbiology
Mitogens - genetics
Mitogens - isolation & purification
Mitogens - metabolism
Molecular Sequence Data
Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
Skin Tests
Streptococcus pyogenes
Streptococcus pyogenes - analysis
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Summary:Streptococcal erythrogenic toxin type A (ET-A) was purified from culture filtrate of Streptococcus pyogenes strain NY-5 grown in a chemically defined synthetic medium NCTC-135. We succeeded in simplifying the purification procedure, and obtained a highly purified preparation of ET-A. The purification procedure was the combination of ultrafiltration with Amicon PM-10 and YM-10 membranes, chromatofocusing with PBE-94 exchanger (pH 4.0-6.0), and gel filtration through Sephacryl S-200. The purified toxin protein showed a single band with Mr 28, 000 on SDS-PAGE and had pI 5.2 on agarose IEF. HPLC chromatography pattern of the toxin revealed one symmetric peak. The result of amino acid analysis of the toxin was in accordance with that of Gerlach et al and with Weeks and Ferretti who reported the nucleotide sequence of the spe A gene. Biological activities of the purified toxin were remarkably potent. The mitogenic activity for rabbit lymphocytes and one skin test dose in rabbit were found at the lower dose of 10pg and 1ng of the toxin, respectively.
ISSN:0385-5600
1348-0421
DOI:10.1111/j.1348-0421.1989.tb01512.x