Open AccessComparison of methods for the production and purification of toxin A from Clostridium difficile
Author(s) -
Torres Javier F.,
Lönnroth Ivar
Publication year - 1988
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1988.tb02569.x
Subject(s) - toxin , papain , chemistry , chromatography , clostridium difficile toxin a , clostridium difficile , polyacrylamide gel electrophoresis , size exclusion chromatography , trypsin , dialysis , microbiology and biotechnology , molecular mass , gel electrophoresis , biochemistry , biology , enzyme , antibiotics , medicine
The production and purification of toxin A from Clostridium difficile were studied. When the toxin was produced in dialysis culture it preicipitated quantitatively at pH 5.5 and after purification it appeard homogeneous in polyacrylamide gel electrophoresis (PAGE). The toxin probably consists of two noncovalently bound peptides, each with a molecular mass of about 250 dDa. It is resistant to trypsin but sensitive to papain and chymotrypsin. In contrast, toxin A produced in anaerbic chamber culture precipitated poorly at pH 5.5 (yield 14%) and easily formed aggregates as observed in gel filtration and PAGE Accordingly, dialysis culture seems to be a better method for producing and purifying toxin A.