Synergistically Teaching Computational and Experimental Approaches to Study Biocatalysis: Kinetic Resolution of Racemic Citronellol with Lipase

Technological advances in computational methods to describe biochemical systems continue to develop at a rapid rate, but cases that use these techniques synergistically with experimentation require more effort. Many research studies involve only dry lab or wet lab techniques but the emphasis of using both approaches together are rare due to the requirement of technical expertise in both areas. A biochemistry laboratory course was taught focusing on enzymatic kinetic resolution, which took advantage of concepts from both computational and experimental approaches. The first six weeks of the semester focused on teaching students dry lab techniques including docking small molecule ligands into the active site of protein structures. The latter half of the course focused on wet lab experiments involving a lipase-catalyzed acetylation of a racemic mixture of citronellol. The assignments during the second half of the semester included developing computational predictions using the free software taught in the beginning of the semester to determine which enantiomer of citronellol (i.e. R-citronellol or S-citronellol) has a higher binding affinity for lipase and matching this theoretical prediction to the experimental results. Experiments that were taught included setting up an enzyme-catalyzed reaction, derivatization of enzymatic products for analysis through thin layer chromatography and NMR spectroscopy. The experiments performed in the latter half of the semester in a problem-based learning manner. The students gained a real-world experience in using both computational and experimental techniques to explore an enzyme-catalyzed reaction.