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Unformatted text preview: Melissa Reyes UF ID: 6786-5269 February 9, 2010 Lab Partners: Sarah, Gabby, Gabby L, Elle Lab Report: Enzyme Kinetics Introduction Enzymes are biological catalysts that are substrate specific, that increase the rate of reaction. Enzymes do not change the nature of the reaction or the final product. Several factors affect an enzymes activity, such as temperature, pH, concentration of cofactors and coenzymes, concentration of enzyme and substrate, and stimulatory and inhibitory effects of products. In this experiment, we used temperature and pH to determine the rate of reactions. This type of experiment would be useful for determining the function of certain enzymes within the body. Certain enzymes work best at certain pHs, thus this experiment could determine the optimal pH for certain enzymes. This experiment could also determine the temperature at which enzymes begin to disfigure or denature. The purpose of this lab is to determine the effects of changes in temperature and pH on the activity of lactate dehydrogenase (mLDH). The reason mLDH is important is because it makes NAD, which is responsible for converting pyruvate to lactate. Methods Concentration of reagents: o 0.2 mM NADH in 54 mM phosphate buffer o 16.2 mM pyruvate in dH 2 O o 1:2000 dilution of muscle homogenate Volumes of reagents: o 0.05 mL pyruvate o 0.9 mL NADH o 0.025 mL homogenate cocktail Gloves, cuvettes, parafilm, and inversion were used to minimize error...
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This note was uploaded on 11/27/2010 for the course APK 2105 taught by Professor Brooks during the Spring '07 term at University of Florida.
- Spring '07