Kinetic Studies on Alkaline Phosphatase from ECHINOPLUTEI1

摘要: Using mass culture of the sea urchin Tripneustes gratilla (L.) eggs, a method was worked out for extracting fairly large quantities phosphomonoesterase alkaline phosphatase. The extracted enzyme showed single ultraviolet absorbance band after elution from Dowex 2 column, and material that absorbed most strongly at this wave length contained It two characteristic bands in starch-gel electrophoresis. When 6.25 mM p-nitrophenyl phosphate hydrolyzed glycine buffer 38C, optimum pH 10.5. hydrolysis conformed to first order reaction, reaction constant 0.00864 min-1. temperature enzymatic action 25–30C, coinciding with range fluctuation animal's ambient temperature. temperature–activity curve close conformity Arrhenius equation, activation energy 11,250 cal/mole. value Km (Michaelis constant) obtained 10.5 (at 25C) is 2.197 × 10-6 M p-NPP/liter.