SC BIOL 101 - Catalase Enzyme Activity (8 pages)

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Catalase Enzyme Activity



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Catalase Enzyme Activity

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Pages:
8
School:
University Of South Carolina-Columbia
Course:
Biol 101 - Biological Principles I
Biological Principles I Documents
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Bio 101L Experiment 3 Catalase Enzyme Activity Report Author Cameron G Kahn Report Submitted 8 October 2014 Data Collected 17 September 2014 Laboratory Partners Nilesh Syam Taylor Hollingsworth Teaching Assistant Amanda Havighorst Author s Affiliation Department of Chemistry and Biochemistry University of South Carolina Columbia SC 29208 I pledge that my work meets the standard of the USC Honor Code ABSTRACT This experiment was performed to show the importance of enzyme activity by examining catalase in the decomposition of hydrogen peroxide Enzymes are important catalysis for chemical reactions that happen within living organisms By studying the effects of different concentrations of substrate and environmental changes to the enzyme the rate of reaction can be evaluated and thus allowing one to create the most ideal situation that will make an enzyme the most efficient in catalyzing a certain chemical reaction The results from performing this lab shows the fastest rate of reaction when the catalase is at a warm temperature and hydrogen peroxide at higher concentrations to a certain extent Upon Statistical analysis of both data sets collected only the results from 0 40 0 20 and 0 10 substrate concentration lied within the 95 confidence interval All other results were affected by potential systematic errors thus causing the data to lie outside the 95 confidence interval 1 INTRODUCTION Within the human body many chemical reactions take place in order for our bodies to function Enzymes are protein catalysts which help in speeding up the rate of a reaction 1 One of the more important enzymes in our bodies is a substance called catalase which strictly deals with the breakdown of hydrogen peroxide H 2O2 into water H2O and oxygen O2 The chemical reaction of the decomposition of hydrogen peroxide with catalase as the catalyst can be seen below 2 H 2 O2 2 H 2 O O2 2 Hydrogen peroxide is considered to be cytotoxic meaning toxic to living cells and therefore it is necessary to break down the substance into water and oxygen To do this catalase is introduced to ensure our survival against the harmful oxidizing effects of H2O2 If an enzyme such as catalase did not exist to help with the detoxification of H 2O2 fatal CO2 bubbles would form in our blood 1 At normal body temperature 37 oC catalase is one of the most efficient enzymes within our body one molecule of catalase can break down 6 million molecules of H2O2 in a single minute 2 Catalase is just one of the many enzymes that control chemical reactions within our body but still contributing an important part towards our overall metabolism and ultimately our survival In this experiment we will be examining the effects on the reaction rate of catalase to detoxify hydrogen peroxide using different concentrations of substrate at room temperature and then different temperatures or conditions of catalase with the concentration of the substrate being constant Our hypothesis is that the rate of reaction will occur the fastest when more concentration of substrate is present due to the saturation of the enzyme up until the active site of catalase is completely full Over saturation would cause the reaction rate to stop increasing and remain constant and eventually a decrease in reaction rate would follow if the over saturation of the enzyme was high enough In the case of temperature change of catalase we believe that we will see the fastest rate of reaction in a warm temperature due to the high efficiency catalase has in the internal environment of our bodies which is at a warm 40 oC and we expect a low rate of reaction when at extremely high temperatures due to the molecule denaturing or low temperatures Denaturing is when the bonds holding the protein structure of an enzyme s specific shape breaks causing the substrate to no longer fit in to the active site of the enzyme Once an enzyme is denatured it is permanently inactive within the reaction This denaturing effect happens at 50 60 oC in the human body METHODS Obtain a 600mL beaker and fill it with water Next use a 10mL graduated cylinder and fill it with water Place a thumb over the opening of the graduated cylinder to create an air tight seal Invert the graduated cylinder and place into the 600mL beaker After making 2 sure there are no air bubbles in the top of the graduated cylinder remove your thumb Obtain a glass tube with a U shaped opening at one end and place the U shaped opening into the inverted 10mL graduated cylinder After the beaker is set up take a 50mL Erlenmeyer flask and add 10mL of substrate H2O2 and 10mL of catalase buffer into the flask Immediately cap off the flask by placing a rubber stopper over the opening and then slide the unused end of the glass tube through the stopper which will subsequently connect both glassware systems together This will complete the set up for the apparatus Once the reaction flask chamber has been capped and connected to the beaker system press start on a stop watch and begin to swirl the reaction chamber containing both the substrate and the catalase Record the time it takes for the 10mL graduated cylinder to fill up with 10mL of oxygen The data collection is complete for that particular trial and the apparatus can be disassembled and reset up for the next trial Make sure to rinse out the Erlenmeyer flask before the next trial begins There will be a total of 9 trials to perform The first trial will be the control and one will only add 10mL of a phosphate buffer along with the 10mL of substrate to the reaction chamber The next 5 trials will test for the effect of different concentrations of substrate on the time it takes for 10mL of oxygen to fill the graduated cylinder The 5 trials consists of 0 8 0 4 0 2 0 1 and then 0 0 of substrate added to the reaction chamber Lastly an additional 3 trials will need to be performed in order to test the effect of temperature of the substrate on the reaction rate for the time it takes 10mL of oxygen to fill the graduated cylinder The three trials will consist of 0 8 of boiled catalase 0 8 warm catalase and 0 8 ice cold catalase All recorded values will need to be written in the chart provided in the lab manual Once the all trials are completed the lab bench and all materials used such as glassware will need to be cleaned and properly stored for safety Consult with the TA for any questions regarding clean up disposal of aqueous solutions and or procedural steps T tests will need to be performed on each data


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