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

Microbiology and Immunology
Y Kamezawa, T Nakahara

Abstract

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 10 pg and 1 ng of the toxin, respectively.

References

Nov 1, 1979·Infection and Immunity·P M SchlievertD W Watson
Oct 1, 1987·Zentralblatt Für Bakteriologie, Mikrobiologie, Und Hygiene. Series A, Medical Microbiology, Infectious Diseases, Virology, Parasitology·D GerlachJ J Ferretti
May 1, 1986·Molecular & General Genetics : MGG·L P JohnsonP M Schlievert
Dec 1, 1974·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·P K Nakane, A Kawaoi
Feb 1, 1980·Infection and Immunity·I van de Rijn, R E Kessler

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Citations

Jul 27, 1999·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·H DobashiI Sekine
Sep 1, 1996·FEMS Immunology and Medical Microbiology·E NemotoK Kumagai
Nov 1, 1996·Clinical and Diagnostic Laboratory Immunology·E M MasciniH van Dijk

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