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A SIMPLE PROCEDURE FOR THE PURIFICATION OF STAPHYLOCOCCAL α ‐TOXIN
Author(s) -
Dalen A. B.
Publication year - 1976
Publication title -
acta pathologica microbiologica scandinavica section b microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0304-131X
DOI - 10.1111/j.1699-0463.1976.tb01948.x
Subject(s) - toxin , chemistry , urea , chromatography , polyacrylamide , electrophoresis , hydrolysis , casein , precipitation , molecular mass , polyacrylamide gel electrophoresis , lysis , biochemistry , enzyme , physics , meteorology , polymer chemistry
Staphylococcal α ‐toxin was produced in a fluid medium based on acid hydrolysed casein using strain Wood 46. α ‐Toxin and several other proteins were precipitated from bacteria‐free culture supernatants by heating at 60° C for 20 min. The process was influenced by the pH of the solution. The toxin was completely inactivated and the precipitate contained a number of proteins if the pH of the solution was adjusted to 4.0–5.0. Heat precipitation of solutions having a pH of 6.0–7.0‐ resulted in a partial inactivation of α ‐toxin. The precipitates at this pH contained less of the additional proteins and had higher relative amounts of α ‐toxin than precipitates formed at a lower pH. The precipitate was dissolved in 8 M urea with the resultant activation of the haemolysin. Pure α ‐toxin with a molecular weight of 39,000 was obtained by electrophoresis in 8 M urea at pH 8.6 in ordinary tubes for polyacrylamide electrophoresis. The separation time was 45 min. The minor component of α ‐toxin with a pI of 7.4 could be demonstrated by the same method. A non‐haemolytic protein with a molecular weight of 27,500 which existed in at least two charged forms, was shown to have an antigenic relationship to the toxin with a molecular weight of 39,000.