Infrared Thermography Applied to the Study of Enzyme Kinetics

Abstract

Kinetic analysis of enzyme activity typically relies on spectrophotometric detection of changes in product or substrate concentrations. Spectrophotometric detection can directly monitor enzyme activity when a suitable chromophore is involved in the reaction itself, or reaction progress can be tracked via a coupled reaction that involves a chromophore with quantifiable signal changes. An alternative, potentially universal strategy for tracking reaction rates involves monitoring the evolution of heat from the enzyme‐catalyzed reaction. The heat generated by a reaction can then be used to calculate the moles of product formed by taking into account the molar enthalpy change for conversion of substrate into product. As with other enthalpimetric approaches, e.g. isothermal titration calorimetry, analysis of the reaction is not dependent on the use of a specific chromophore, nor is it challenged by optically‐dense or heterogeneous samples. Such methodologies have been in use for decades using near‐adiabatic instrumentation to quantify the total heat change. More recently, infrared imaging technologies such as the 200USD FLIR ONE camera open new opportunities to analyze kinetics on the basis of the heat evolved by the enzyme‐catalyzed reaction. We report our preliminary work to demonstrate the proof‐of‐principle use of infrared thermography to track enzyme reaction rates. In addition, the unique advantages of IR imaging are well suited for applications of this approach in a biochemistry laboratory course setting.Support or Funding InformationNational Science Foundation DUE Award Number:1813313; “Collaborative Research: Visualizing Chemistry with Infrared Imaging”This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.