Abstract
The application of immobilized enzymes in flow microcalorimetry using a modified flow cell containing the enzyme preparation is described. The substrate was either injected into the system or applied as continuous flow. The sensitivity of the calorimetric response was studied. For example, injection of only 1 μmole of the substrate benzoylarginine ethyl ester in Tris-HCl buffer, produced an integrated heat response of about 60 mJ when passing through the flow cell containing immobilized trypsin. The enzyme was kept entrapped in uniform-sized spherical polyacrylamide beads showing extremely low flow resistance when packed in the microcolumn, thus giving stable base lines. The potentials of the combination immobilized enzymes-microcalorimetry as analytical tool are discussed.
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