:Immobilization of Enzyme by Radiation-copolymerization and Its Application to Analysis Part II

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Pyruvate kinase (PK) was immobilized by the radiation-copolymerization method at a low temperature. From the mixture of PK solution (1ml), 30% acrylamide +2.7% N, N'-methylene bisacrylamide (1ml), 30% magnesium acrylate (2ml) and 30% sodium acrylate (1ml), the immobilized PK was prepared with good physical qualities and high retained activity of more than 80% of the original. Comparing with native PK the immobilized enzyme was more stable against heat and pH change. Immobilization decreased the Km values both for PEP and ADP, and shifted the optimum pH from 7.5 to 8.0. The immobilized PK was subjected to a 30 min. reaction repeatedly up to 20 times with little activity loss. Determination of ADP and hexokinase activity were successfully performed by use of the immobilized PK.
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