Microcalorimetry as a tool in enzymatic kinetics research

Abstract

Enzymes, as biocatalysts, are an important target of many therapies and are also of great industrial importance, which is why repeatable and accurate parameterization of enzymatic catalysis is very important. The most popular spectrophotometric detection method in enzymology, despite its low cost and speed, often cannot be used directly due to the inappropriate spectral properties of substrates and products. It is then necessary to use auxiliary enzymes, chemical modification of substrates or post-reaction analysis, which may increase the cost of measurement, extend its time or affect the accuracy. Isothermal titration calorimetry is a method widely used mainly for the characterization of inter-molecular interactions, however, its use in enzyme kinetics is gaining more and more recognition due to the direct measurement of the reaction rate using the universal parameter of heat, high sensitivity and low reagent consumption. This work discusses two strategies for conducting a kinetic calorimetric experiment and their applications.