Cell Lysis Optimization to Measure Catalase Activity

Catalase is an enzyme that is found in almost all living organisms that are exposed to oxygen. The purpose of catalase is to break down hydrogen peroxide into its two components, water and oxygen. This process serves a vital role to protect cells from oxidative damage caused by hydrogen peroxide. Catalase is found among many organisms, including bacteria. Escherichia coli (E. coli) is a type of bacteria that is naturally found among the human digestive system where it aids in digestion and vitamin production. This study is focused on optimizing cell lysis for E. coli for catalase retrieval. Different methods of cell lysis include freeze-thaw lysis, thermal lysis, and sonication. Each of these methods were performed along with a colorimetric assay to determine catalase presence. When catalase activity is inhibited it produces a more intense color change as compared to uninhibited catalase which has little color. The intensity of the color reaction was measured at maximum wavelengths of 440 nm and 640 nm. Through this experiment, thermal lysis was shown to be the most effective and can potentially be more effective with optimization such as temperature and time. Future directions include using this E. coli-derived catalase in combination with food-based flavonoids to test inhibition. This inhibition through the consumption of high-flavonoid foods can be beneficial to the body under some pathological conditions.