Pshezhetskiĭ A V, Kovalysheva G V, Chernoglazov V M, Levashov A V, Kliachko N L
Bioorg Khim. 1989 Apr;15(4):437-43.
Micellar solutions of surfactant in organic solvents with rubber additions are proposed for determination of active enzyme concentration. A kinetic theory of enzymatic reactions in reversed micellar systems is developed, suggesting the intermicellar transport of the substrate to be the limiting step in viscous medium. Under these conditions, it is shown that fraction of the product formed after quick transformation of the substrate located in the enzyme-containing micelles depends upon active enzyme concentration and aggregation number of surfactant molecules. The proposed approach is used for the active-site titration of trypsin and cellobiase and for the determination of the aggregation number of Aerosol OT (AOT) molecules in the ternary system AOT/water/cyclohexane.
提出了在添加橡胶的有机溶剂中制备表面活性剂胶束溶液来测定活性酶浓度。建立了反胶束体系中酶促反应的动力学理论,表明底物在粘性介质中的胶束间传输是限速步骤。在这些条件下,结果表明,位于含酶胶束中的底物快速转化后形成的产物分数取决于活性酶浓度和表面活性剂分子的聚集数。所提出的方法用于胰蛋白酶和纤维二糖酶的活性位点滴定以及三元体系AOT/水/环己烷中气溶胶OT(AOT)分子聚集数的测定。