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KMID : 0357319850200020199
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Journal of the Korean Society for Microbiology
1985 Volume.20 No. 2 p.199 ~ p.211
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Purification of Heat-Stable Enterotoxin from Enterotoxigenic Escherichia coli
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Abstract
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Heat-stable enterotoxin (ST) from enterotoxigenic E. coli 0148H28(ST , LT+) that is pathogenic for human was mass-produced in a chemically defined succinate salts medium. The culture supernatant (crude S1) was purified by sequentially applying batch adsorption chromatography on Amberlite XAD-2 resin, ion exchange chromatography on DEAE-Sephacel anion exchanger, gel filtration chromatography on Sephadex G25 and preparative polyacrylamide slab gel electrophoresis. About 60-fold purification was achieved with a yield of about 15% and minimum effective dose of this purified ST was about 3.4 ng in infant mouse assay. Homogeneity of purified ST was demonstrated by showing a single band in analytical SDS polyacrylamide disc gel electrophoresis. Heat-stability of purified ST was tested and 4 mouse units of ST activity was completely lost by treating at 100°C for 30 min. Antiserum obtained from BALB/c mouse immunized with ST-BSA conjugate neutralized the toxic activity of purified ST.
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