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cec's lab report

cec's lab report - The effects of temperature pH...

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The effects of temperature, pH, substrate-concentration, enzyme-concentration, potassium molybdate, inorganic phosphate, and phenyl phosphonate on the rate of an alkaline phosphatase enzyme-catalyst reaction Abstract The experiment required the use of alkaline phosphatase, an enzyme used to increase the reaction rate for an enzyme-cataly zed reaction. The first part of the experiment was used to determine the semi-quantitative estimates of enzyme activity. The potassium molybdate decreased the rate of the reaction, as did the inorganic phosphate and the phenyl phosphate. The enzyme was tested at different pH levels, ranging from four to twelve. At 0 o C, 25 o C, 37 o C, and 100 o C, the rate of absorbency was recorded to determine optimum temperature for the enzyme to aid in the reaction. Different levels of substrate and enzyme concentration were added to the tubes to determine the effect of different concentrations on the rate of the reaction. The more alkaline phosphatase added to the reaction, the faster the chemicals reacted; the same was true for the additional substrate. The results imply that the enzyme prefers a basic solution, in a specific environment, with high amounts of substrate and enzyme. Introduction In laboratory four, the objectives were to understand the effects of inhibitors, temperature , and pH , as well as the enzyme and substrate concentration on enzyme-catalyst reactions (Bill Davis et al. 2007). There were four different experiments preformed to acquire the answers to these questions: how does the addition of other materials effect the enzyme-catalyst reaction, how do the enzyme or substrate concentrations affect the enzyme-catalyst reaction, how does the pH of the buffer affect the rate of the enzyme-catalyst reaction. An enzyme is “an organic catalyst (usually a protein) that accelerates a specific chemical reaction by lowering
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the activation energy required for that reaction, (Soloman et al. G-15). The enzyme being studied in the laboratory was alkaline phosphatase , which is “purified from a species of shrimp, Pandalus borealis ,” (Bill Davis et al. 2007). Another substance tested was substrate, “a substance on which an enzyme reacts; a reactant in an enzymatically catalyzed reaction,” (Soloman et al. G-42). Each time an experiment was conducted the absorption rate was measured for the product, a “substance formed by a chemical reaction,” (Soloman et al. G-36). The effects in each part of the lab were affected by the potassium molybdate, phenyl phosphonate, and inorganic phosphate, which can be construed as inhibitors, substances that block the reaction. Since the potassium molybdate, inorganic phosphate, and phenyl phosphonate were labeled as inhibitors, it was assumed they would slow down the reaction rate. The potassium molybdate is a competitive inhibitor, due to the fact it changes the composition of the alkaline phosphatase in the reaction. The inorganic phosphate slows down, actually reverses, the reaction because it is an excess in the product of the reaction. The third inhibitor added was phenyl phosphonate
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cec's lab report - The effects of temperature pH...

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