| Abstract | Alkaline Phosphatase (ALP) are enzymes that are primarily found in E. coli bacteria, however, it is also found in most eukaryotes and prokaryotes. An example of a eukaryote that carries alkaline phosphatase are human. In humans, alkaline phosphatase plays many significant roles such as fighting bad bacteria in the gut, providing strength to bones and teeth, improving digestion, and sustaining cholesterol levels in pregnant women. Therefore, in this experiment, we are looking at the alkaline phosphatase as it is significant for humans and observing the effects of two inhibitors, L-phenyl alanine, and sodium phosphate, on its activity. We hypothesize that using L-phenyl alanine and sodium phosphate with the reaction of alkaline phosphatase enzyme and PNP substrate will reduce the activity of the alkaline phosphatase enzyme. To test our hypothesis, we first found out the best concentration of the enzyme needed for this reaction by keeping the substrate concentration constant and changing the enzyme concentration while collecting kinetics of the enzyme and plotting it on excel. After finding the best concentration of the enzyme, we shifted to producing a Lineweaver graph of the enzyme-substrate reaction without inhibitor by trying different concentrations of the substrate with a constant concentration, which is the best concentration, of the enzyme, collecting the kinetics data, and plotting them on excel. Finally, on the same graph, similar steps were repeated but with the addition of inhibitors, each one at a time, to check the effects of the inhibitors of the kinetics of alkaline phosphatase. As a result, our hypothesis was proved based on the collected data and both inhibitors seem to be competitive inhibitors.
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