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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
Author(s) -
Kamezawa Yukio,
Nakahara Takako
Publication year - 1989
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1989.tb01512.x
Subject(s) - chromatofocusing , toxin , streptococcus pyogenes , biology , microbiology and biotechnology , size exclusion chromatography , sepharose , streptococcus , ultrafiltration (renal) , chromatography , biochemistry , bacteria , chemistry , staphylococcus aureus , genetics , enzyme
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 M r 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.