Investigating Inhibitors of a Phosphatase Enzyme

Alkaline phosphatases are enzymes responsible for dephosphorylation of phosphate monoesters. They are involved in numerous biological processes but play a particularly important role in bone development and liver function. Abnormal levels of these enzymes can have detrimental effects on the corresponding processes. This study involved enzyme kinetic studies on bovine alkaline phosphatase to test the effects of substrate and enzyme concentrations, pH, and the addition of known and novel inhibitors. Increased substrate and enzyme concentrations increased the rate of catalysis and allowed for the determination of kinetic constants; Km and Vmax. The optimal pH was determining to be approx. 10, and from comparison of rates of catalysis between uninhibited and inhibited assays, the mode of inhibition was determined to be competitive for the known inhibitors: EDTA and phosphate. Several novel thiazole-based inhibitors with different substitution patterns were also synthesized and tested for their efficacy. Department: Biology Faculty Mentor: Dr. Tina Bot