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Keywords:

  • Instrumentation;
  • fluorescence;
  • isothermal titration calorimetry;
  • lysozyme

Abstract

Our first-semester biochemistry course is accompanied by a project-based laboratory with a focus on the enzyme lysozyme. The first half of the semester involves purification of the enzyme. In the second half, students learn to work with advanced instrumentation techniques, specifically fluorescence spectroscopy and isothermal titration calorimetry. These methods are linked, as much as possible, to the theme of lysozyme: its structure, stability, or binding of ligand. As a final project, students design an experiment involving one of these methods. Examples are determination of the pKa of the catalytic residue and the binding affinity for an inhibitor under different conditions of pH and temperature. Four weeks of laboratory are sufficient for students to learn how to use the instruments and to develop a short project. At the end of the project, students give an oral presentation on the theory of the method and their results and prepare a paper that undergoes peer review. Hands-on experience with these methods reinforces theoretical concepts taught in the lecture portion of the course.