| Abstract | Alkaline phosphatase is a metalloenzyme that hydrolyzes phosphomonoesters. The activity of alkaline phosphatase is dependent on the environment and factors that are present within a reaction. L-phenylalanine acts as an uncompetitive inhibitor of AP at concentrations equal to 5mM or more. This project studies the kinetic parameters—KM and VMAX—of the enzyme in the presence of L-phenylalanine as well as the mechanism of inhibition by performing an enzyme activity assay. P-nitrophenyl phosphate was used as a substrate, which produces inorganic phosphate and p-nitrophenol upon hydrolysis. P-nitrophenol is yellow coloured and absorbs at 410 nm, thus, product formation was quantified by measuring absorbance at 410 nm. We hypothesized that L-phenylalanine would uncompetitively inhibit Alkaline phosphatase activity, i.e., a decrease in VMAX and KM values was expected. A better understanding of inhibition mechanisms can then be used towards developing drug therapeutics against diseases.
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