Enzymes are Nature's catalysts and often perform very challenging reactions in a highly selective manner. However, accessing and purifying enzymes for synthetic purposes can be challenging and expensive, limiting their use by organic chemists. As an alternative, we can add cells directly to our reactions which harbor the enzyme of interest. Monitoring the progress of a reaction in this format (called whole cell reactions) can also be a challenge, as the cells must first be removed to check reaction progress. In this project, we developed a new spectrophotometric method by which whole cell reactions can be monitored without cell removal. This method uses an integrating cavity spectrophotometer to quantify reaction progress live, offering a streamlined solution to this challenge. We performed kinetic profiling experiments to analyze the reaction of a ring-cleaving dioxygenase in both whole cell and purified enzyme format. We evaluated differences in enzyme behavior in these different preparations. The results of these experiments and the advantages of this approach will be presented.
