Lab_Report_Enzyme_Catalysis

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Apr 3, 2024

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Enzyme Catalysis Danielle Fouche 3/13/2024

1 © 2016 Carolina Biological Supply Company Instructions: 1. Please read all of the introduction and background information within the investigative manual. a. Once you have done so answer the prelab question BEFORE completing any of the lab’s activities. 2. Once you have completed the prelab questions proceed to the activities of the lab within the investigative manual. a. As you read through the instructions for completing each activity make sure you also: i. Complete any instructions (append photos, etc)/ and answer any questions found in the post lab questions for each activity. ii. Take the photos of your experiments in each activity as directed below . IMPORTANT : Don’t clean -up your lab until you know what portion of the experiment you need to take a picture of. 3. Here is a video that will introduce you to the lab and its main concepts. The student is encouraged to watch it. a. Enzyme Catalysis Prelab Questions 1. Define activation energy. Will activation energy for a reaction be higher or lower when an enzyme is present to catalyze it? Why? The lowest amount of energy required to start a chemical reaction is activation energy. When an enzyme is present to catalyze it, the activation energy is lower. Enzymes bind to a substrate and hold it in a way that lets the reaction happen more effectively. ( Enzymes Review (Article) , n.d.) 2. There are a number of conditions that will impact the rate an enzyme is capable of catalyzing a reaction including: temperature, pH, enzyme concentration, and substrate concentration. In this lab you will investigate the effects of temperature and enzyme concentration on the rate of an enzyme. How though will substrate concentration affect the rate of an enzyme? To investigate this, google “How does substrate concentration affect enzyme activity?” and answer the questions below: a. Does an increase in substrate concentration lead to an increase in enzyme activity?

2 © 2016 Carolina Biological Supply Company An increase in substrate concentration will lead to an increase in enzyme activity at first, but as the enzymes are completely saturated the activity will no longer increase because the substrate has nothing left to bind to. ( Enzymes Review (Article) , n.d.) b. Are there limitations to the answer you gave in “a.”? In other words, can an increase in substrate concentration ever NOT lead to a detectable increase in activity of the enzyme? Why or why not? Yes, there are limitations. The activity will only increase with the increase in substrate concentration initially until all the enzymes are saturated. If all the enzymes are saturated then there is nothing for the substrate to bind to, therefore the enzyme activity will not increase but it will remain the same. ( Enzymes Review (Article) , n.d.) 3. As stated above, in this lab we will be investigating the effect of enzyme concentration (activity 1) and temperature (activity 2) on the rate of an enzyme. BEFORE you begin each activity please fill out the purpose and hypothesis statements found below. Activity 1 Instructions: 1. Open the investigative manual. Locate all the needed materials supplied in the kit and those you will need to supply yourself. 2. Lay them out in your work area. 3. Read through the entire set of instructions found in the investigative manual for the activity to avoid making mistakes when you go to execute the experiment. 4. Once you have read through the instructions go back to step 1 and begin executing the experiment. 5. Please answer the questions below and/or append appropriate representations of data (photos, graphs, etc). REMEMBER don’t clean up until you have taken the appropriate photos of your experiment as described below. Purpose statement: How does the substrate concentration affect the reaction time or activation energy?

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3 © 2016 Carolina Biological Supply Company Hypothesis statement: If the substrate concentration is higher than the enzyme activity will initially increase but will then remain unchanged after a prolonged amount of time. Evidence/Claim statement: According to the results of this experiment the higher the substrate concentration the higher the enzyme activity was. With higher concentration levels of the catalysis on the paper the time that it took for the paper to reach the top of the hydrogen peroxide/water solution was lessened. Reflection statement: The reaction of the paper floating to the top of the solution was able to happen more efficiently the higher the substrate concentration was because the enzymes require a lower activation energy. Photo 1 – Activity 1 Take a picture and insert the image(s) of your medicine cup after step 8. of the “ Enzyme Concentration ” section in activity 1 of the investigative manual:

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7 © 2016 Carolina Biological Supply Company Data Table 1 0% 25% 50% 75% 100% Depth of H 2 O 2 Solution (From step 11.) 20ml 20ml 20ml 20ml 20ml Trial 1 Time 180 seconds 7.64 seconds 5.63 seconds 4.98 seconds 2.57 seconds Trial 2 Time 180 seconds 10.15 seconds 4.29 seconds 4.41 seconds 3.05 seconds Avg. Time 180 seconds 8.90 seconds 4.96 seconds 4.70 seconds 2.81 seconds Rate ( d / t ) (From step 12.) 0.00 mL/sec 2.25 mL/sec 4.03 mL/sec 4.26 mL/sec 7.12 mL/sec Graph 1 – Activity 1 Insert your graph from step 13 . of the “Enzyme Concentration” section in activity 1 of the investigative manual:

8 © 2016 Carolina Biological Supply Company 1. Having now completed activity 1 and your graph go back and fill in your Evidence/Claim and Reflection statements and then answer the question(s) below. 2. Based on the information in Data Table 1 and your graph, explain the relationship between percent concentration of catalase and reaction rate. From this what can we conclude about the effect that enzyme concentration has on the rate of a reaction? Based on the graph the higher the percent concentration of catalase the faster the reaction rate was. 3. Using the graph, predict the rate if catalase concentrations are 10%, 60% and 85%. Using the graph, my prediction would be that the reaction rate for 10% would be faster than 0% but slower than 25% at 1.10 mL/sec. The 60% would be a quicker reaction than 50% but slower than 75% at 4.10 mL/sec. The 85% would be faster than 75% but slower than 100% at 5.7 mL/sec. 0 1 2 3 4 5 6 7 8 0 25 50 75 100 Rate Percent Concentration of Enzyme Enzyme Concentration

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9 © 2016 Carolina Biological Supply Company Activity 2 Instructions: 1. Open the investigative manual. Locate all the needed materials supplied in the kit and those you will need to supply yourself. 2. Lay them out in your work area. 3. Read through the entire set of instructions found in the investigative manual for the activity to avoid making mistakes when you go to execute the experiment. 4. Once you have read through the instructions go back to step 1 and begin executing the experiment. 5. Please answer the questions below and/or append appropriate representations of data (photos, graphs, etc). REMEMBER don’t clean up until you have taken the appropriate photos of your experiment as described below. Purpose statement: How will temperature affect the activation energy? Hypothesis statement: If the temperature is warmer than the reaction time will be faster and activation energy will be lower than if the temperature is colder. Evidence/Claim statement: The hypothesis was supported in that the higher the temperature was the lower the activation energy was, which is why the paper floated to the top of the solution quicker with each temperature increase. Reflection statement: I learned the not only does changing the solution concentration affect the activation energy, but so does the temperature. This makes me wonder what other circumstances might change the activation energy.

10 © 2016 Carolina Biological Supply Company Data Table 2 10 deg. below 15 o C Room temp. 25 o C 10 deg. Above 35 o C 20 deg. Above 45 o C 30 deg. Above 55 o C Depth of H 2 O 2 Solution 20mL 20mL 20mL 20mL 20Ml Time 4.64 seconds 3.41 seconds 1.11 seconds 1.17 seconds .97 seconds Rate ( d / t ) 4.31 mL/sec 5.87 mL/sec 18.02 mL/sec 17.10 mL/sec 20.62 mL/sec Graph 1 – Activity 2 Insert your graph from step 17 . of the “ Effect of Temperature ” section in activity 2 of the investigative manual: 1. Having now completed activity 2 and your graph go back and fill in your Evidence/Claim and Reflection statements and then answer the question(s) below. 0 5 10 15 20 25 15 25 35 45 55 Rate Temperature In Degrees Celcius Effects of Temperature on Enzymes

11 © 2016 Carolina Biological Supply Company 2. Based on the information in Data Table 2 and your graph, Is there an optimum temperature for catalase that makes it the most productive in terms of reaction rate? At what temperature does the rate decrease? If your data does not show a decrease do you think there is a temperature in which rate would begin to decrease? Explain why. Out of the temperatures observed the ideal temperature for faster activation energy was 55 degrees Celsius. From the temperatures observed the colder temperatures are the ones that had a decreased rate. The reason for this is because the higher the temperature is the faster the molecules move which causes them to collide more frequently. This causes the activation energy to be lower.

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12 © 2016 Carolina Biological Supply Company Carolina Biological Company. (2016). Cell structure and function: eukaryotes, prokaryotes and viruses. investigation manual and lab Enzymes review (article) . (n.d.). Khan Academy. https://www.khanacademy.org/science/ap-biology/cellular- energetics/environmental-impacts-on-enzyme-function/a/hs-enzymes- review#:~:text=Enzyme%20structure%20and%20function&text=Enzymes%20l ower%20the%20activation%20energy

Lab_Report_Enzyme_Catalysis

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