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Effect_of_pH_and_Temperature_on_Enzyme_Activity - Karim 1...

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Karim 1 Effects of pH and Temperature on Activity of Alkaline Phosphatase Abstract : In order to determine different effects of pH and temperature on the activity of enzymes, an assay was conducted at multiple temperatures and pH levels with an enzyme, alkaline phosphatase. The substrate used was p-Nitro-Phenyl Phosphate, pNPP. In doing so, optimal conditions, or the most favorable conditions of enzymes, could be observed. The absorbance of the solutions was recorded using a Spectronic 20 Colorimeter at 415nm, which proved the enzyme was most efficient in high temperatures and high pH levels, over 7.0. The highest absorbance was recorded at 37 C and pH of 10. These results disprove the hypothesis that it works best at high pH values and high temperature values. It actually works best at basic conditions and at temperature near 37 C, a normal body temperature. Introduction : Enzymes are biological catalysts, having high importance in biological systems. Enzymes speed up reaction rates and catalyze chemical reactions, lowering the activation energy needed to initiate a chemical reaction, without affecting the change in free-energy. The activation energy is the energy required to break bonds and start a reaction. Enzymes form an intermediate enzyme- substrate complex, which is a linkage between the enzyme and a substrate during a chemical reaction. The enzyme binds to the substrate, a substance which it acts upon. The enzymes contain active sites, where the substrate binds, to form the enzyme-substrate complex. Products are released when the complex is broken up, and the original enzyme can be restored (Solomon et al ., 2011). Factors such as pH and temperature, affect the rate of an enzyme-catalyzed reaction (Davis, et al., 2011).
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2 Alkaline phosphatase is an enzyme usually found in the liver and bones. It is also found in the intestines and kidneys (Healthwise Staff, 2010). It is an important enzyme that plays a role in catalyzing phosphate within living cells. Alkaline phosphatase can convert p-Nitro-Phenyl Phosphate (pNPP), the substrate, to p-Nitrophenol (pNP) and inorganic phosphate (Pi) in a reaction that can also be reversed (Davis, et al., 2011). This experiment will test the hypothesis that the enzyme, alkaline phosphatase, is most efficient when pH and temperature are in effect at their optimal conditions, at a high temperature and a high pH. It is possible that the reaction rate may slow down or stop if the conditions are unfavorable. If the pH is too acidic, it will cause the protein to denature, or lose its three-dimensional shape. The higher the pH of a substance, the more effective the enzyme will be because of the increase in the amount of hydrogen ions. If the optimal conditions for temperature are favorable, there will be an increase in molecular motion and increase the rate of the enzyme. If the temperature is too high, the enzymes will reach a certain point where it will begin to denature and change the molecular confirmation. Materials and Methods
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Effect_of_pH_and_Temperature_on_Enzyme_Activity - Karim 1...

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