CHARACTERIZATION AND DISTRIBUTION OF AMYLASES DURING VEGETATIVE CELL-GROWTH AND SPORULATION OF CLOSTRIDIUM-PERFRINGENS

Clostridium perfringens produced eight extracellular and two intracell ular amylolytic activities when examined by zymograms following polyac rylamide gel electrophoresis under native conditions. The major intrac ellular amylase was isolated from vegetative cells of C. perfringens. It possessed an estimated molecular mass of 112 kDa. Sulfhydryl and ph enol functional groups were essential to its activity. The amylase was endo-acting on stanch and also hydrolyzed pullulan. Polyclonal antise ra against a purified extracellular amylase did not cross-react with i ntracellular amylase and the two amylases were biochemically different . The distribution of extracellular amylolytic activities of sporulati ng cells was different from thar of vegetative cells, whereas tile dis tribution of intracellular amylolytic activities remained identical. A significant increase of a particular amylase (AS) occurred in the ext racellular fluid during sporulation compared with that during vegetati ve growth. Regulation of the excretion of amylase(s) may be sporulatio n and enterotoxingenicity related.