福島, 和雄

田村, 五郎

FUKUSHIMA, Kazuo

TAMURA, Goro

(1) The lotus seed protease showed the maximum activity at pH 4.0 for 6M urea denatured casein. The degree of hydrolysis for this urea denatured substrate was higher than that for heat denatured substrate (about 4 times). Accordingly, the enzyme assay system, useing urea denatured casein as the substrate, was employed in order to determine the proteolytic activity. (2) The partially purified preparation, obtained by dialysis and pH treatments, and fractional precipitations with ammonium sulfate and ethanol, showed relatively higher purification degree than that of the Sephadex G-100gel filtration state reported in the previous paper. (3) In order to obtain further purified preparation, the purification procedures of chromatography and batch separation on DEAE cellulose, Sephadex-G-100 and G-200gel filtrations, CMcellulose column chromatography, and starch zone electrophoresis, were examined, By uses of DEAE cellulose batch separation and electrophoresis, about 5 folds increase, in specific activity, compared with the partially purified preparation, was obtained. It seems, however, that this purified enzyme was not so homogenous on starch electrophoresis and also on CM cellulose column chromatography.