Engineering of Catalytic Activity into THI3 Protein

Indianapolis, IN

Serine hydrolases are a broad class of enzymes that catalyze a range of biological and chemical reactions. With over 100 serine hydrolases present in humans, the biological functions of many hydrolases are still undiscovered. The goal of my research was to measure the hydrolase activity in the simple eukaryotic organism, Saccharomyces cerevisiae, and to begin to assign the chemical reactivity of all serine hydrolases in yeast. A library of fluorogenic hydrolase substrates with diverse chemical structures was incubated with S. cerevisiae cultures. The relative activation of the fluorophores by serine hydrolases was then compared across various growth conditions, including in the presence of general serine hydrolase inhibitors. In future work, yeast deletion strains for serine hydrolases will be used to assign the measured enzyme activity to specific serine hydrolases and potential biological reactions in yeast.