Proteoytlic enzyme of the stomach of
Tilapia nilotica was purified by ammonium sulfate precipitation, followed by ion-exchange chromatography (DEAE-cellulose), chromatofocusing (polyexchanger PBE 94) and gel filtration (Sephadex G-100). The enzyme was found to be in pure forms when examined by electrophoresis.
The specific activity of the purified enzyme was 22 times that of the crude extract.
The protease had a molecular weight of 54, 000, and showed the highest activity at pH3.5 and 50°C. This was stable at pH3.5-5.5 and below 50°C.
The
Km value of the enzyme for hemoglobin was calculated to be 5.4mg/ml.
The enzyme activity was inhibited by 1, 2-epoxy-3-(
p-nitrophenoxy) propane, diazoacetyl-DL-norleucine methyl ester, and pepstatin.
The effect of various inhibitors on enzyme activity was examined. As a result, it was presumed that this enzyme may be classified into an asparatic protease. The enzyme specifically digested carbobenzoxy-glycyl-tyrosine, carbobenzoxy-glutamyl-tyrosine, benzoxy-glycyl-phenylalanine, and carbobenzoxy-glutamyl-phenylalanine.
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