Figure 2

Thermally induced inactivation of free and immobilized preparations of Tv DAO. Time courses of inactivation of TvDAO immobilized on Strep-Tactin MacroPrep material (open squares) and Sepabeads EC-EP (full circles) are compared to the time course of inactivation of the soluble enzyme (full triangles). The inactivation time course of the Sepabeads immobilizate in the presence of 100 μM FAD is shown as open circles. The enzymes were incubated at 50°C on an Eppendorf Thermomixer using orbital shaking at 500 rpm; the protein concentration was 1 μM in 10 mM Tris-HCl buffer, pH 7.5 in all cases. Solid lines represent least squares fits of a single exponential decay function to the data. Note that the suspensions of immobilized biocatalysts were handled like liquids, which explains the observed scattering of the data for the enzyme immobilized on Sepabeads EC-EP.