ポリアクリルアミドゲルによるサッカラーゼの不溶化とその性質

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An active water-insoluble saccharase was prepared by means of entrapping a soluble saccharase in a polyacrylamide gel and the enzyme kinetics of the insoluble form were compared with those of the soluble form. Moreover, a continuous hydrolysis of sucrose was carried out by using a column packed with the insoluble form.The activity of the insoluble form retained was 8〜10% of that of the soluble form and there was no significant difference between the two forms of saccharase with regard to the influence of variations in pH and inhibitor concentration. Considerable differences existed between the two forms of saccharase in their behavier toward temperature. The soluble form was no longer active at 65℃, but the insoluble form still retained 8% of its activity at 70℃. And when the two forms of saccharase were treated for 30 min at a specified temperature, the insoluble form had no activity at 60℃, and the isoluble form retained 15% of its activity. However, there was no difference between the two forms of saccharase with regard to the optimum temperature and the apparent activation energy. Michaelis constants were same for the two forms of saccharase, but the maximum rate of sucrose inversion for the insoluble form was decreased to about 1/10 that for the soluble form.The action mechanism of the insoluble form in a column was studied by using a modified form of the integrated Michaelis-Menten equation. As a result, the values of the apparent Michaelis constant of the insoluble form were shown to be dependent on the flow rate in the column, but the values of the rate constant were shown to be independent of the flow rate.
社団法人日本生物工学会の論文
1973-11-25

ポリアクリルアミドゲルによるサッカラーゼの不溶化とその性質

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