Measuring Enzymatic Stability by Isothermal Titration Calorimetry

J Vis Exp. 2019 Mar 26:(145). doi: 10.3791/59302.

Abstract

This work demonstrates a new method for measuring the stability of enzyme activity by isothermal titration calorimetry (ITC). The peak heat rate observed after a single injection of the substrate solution into an enzyme solution is correlated with enzyme activity. Multiple injections of the substrate into the same enzyme solution over time show the loss of enzyme activity. The assay is autonomous, requiring very little personnel time, and is applicable to most media and enzymes.

Publication types

  • Video-Audio Media

MeSH terms

  • Calorimetry / methods*
  • Catalysis
  • Enzyme Stability
  • Humans
  • Kinetics
  • Lactase / chemistry*
  • Lactase / metabolism*
  • Substrate Specificity
  • Thermodynamics
  • beta-Fructofuranosidase / chemistry*
  • beta-Fructofuranosidase / metabolism*

Substances

  • Lactase
  • beta-Fructofuranosidase