The purpose of activity C is o determine the rate at which H2O spontaneously decomposes when exposed to room temperatures and ambient light for 24 hours. The purpose of activity D is to determine the rate at which catalane decomposes H2O. After adding HOSTS for different time lashes, etc. , the resulting data will be graphed at which the catalane decomposed by catalane. Background: The four different activities to the enzyme catalyst lab have similar but different backgrounds. Activity Ass background is to investigate the specific reaction of the decomposition of hydrogen peroxide by the enzyme, catalane.
Hydrogen Peroxide decomposes lowly into water and oxygen, and the addition of catalane lowers the activation energy of the reaction until it proceeds to a room temperature. The catalane will is unchanged and is usable for to catalyst the reaction of more hydrogen peroxide. The background of activities A, B, and C is to determine the concentration of H2O in a solution. To do this, the concentration of an unknown will be used, such as a solution of 0. 1 N, which is normal. When titration is done, 5 ml of acid will go in a beaker. When pH indicator paper is added, the paper will turn yellow if it is an acid and green if the solution is neutral.
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This is the endpoint of titration. In the activities, 2% potassium permanganate will be used to titrate H2O. When the H2O reacts with the potassium permanganate, which is dark purple, the solution will become persistent or neutral pink or pale light brown. This ends the titration for the part. Materials: Four H2O in labeled cup Three HOSTS in labeled cup Catalane on ice Three Km Unlabeled 60 ml cup Test tubes Test tube holders Test tube racks Four syringe labeled H2O Transfer pipettes Glass rods Scalpels Potato Cube Hot plate Cup labeled Baseline Two Cup labeled titration Syringe labeled HOSTS
Syringe labeled transfer Titration syringe Distilled water in cup labeled 20th Cup labeled unconcealed decomposition Cup labeled H2O overnight Cups labeled respectively 10 sec, 30 sec, 60 sec, 120 sec, and 180 sec. Procedure: Activity A 1 . With the syringe labeled H2O and transfer 10 ml of H2O into the unlabeled 60 ml cup. Add 1 ml of catalane to the unlabeled cup. Observe. 2. With a transfer pipette, transfer 5 ml of catalane to a test tube. Place the test in water filled beaker on the hot plate, and let it boil. 3. Transfer 10 ml of H2O into clean cup. Add 1 ml of catalane to the cup. Observe.
Wash out beaker. Cut a potato cube about LLC on the sides. Place the cube in an unlabeled cup and crush with a glass rod. Don’t use too much force as the rod will break. Activity B 1. Use H2O syringe and put 10 ml of H2O in 60 ml cup labeled baseline. 2. Add 1 ml of distilled water from 20th cup to baseline cup with transfer pipette. 3. Use H2O syringe and add 10 ml of 1 M HOSTS from HOSTS cup to baseline cup. 4. Mix contents of baseline cup by swirling the cup. 5. Use 5 ml syringe labeled transfer and remove 5 ml of baseline cup solution and put it in the cup labeled titration. Rinse the syringe afterwards. Titrate the 5 ml sample to determine the baseline amount of H2O. Activity C 1 . Use H2O syringe and transfer 10 ml of H2O from the H2O overnight cup to the 60 ml cup labeled unconcealed decomposition. 2. With the transfer pipette, add 1 ml of distilled water from the 20th cup to the unconcealed decomposition cup. 3. With the 10 ml syringe labeled HOSTS, add 10 ml of M HOSTS from HOSTS cup to the unconcealed decomposition. 4. Swirl the unconcealed decomposition cup to mix the contents. 5. With the 5 ml syringe labeled transfer, remove 5 ml of the reaction mixture and put it in the titration cup. Rinse the transfer syringe. 6.
Titrate the sample to determine the amount of H2O left in the solution after 24 hours. Activity D 1. Line up the 60 ml cups with the labels 10 sec; 30 sec, 60 sec, 120 sec, and 180 sec. Use the H2O syringe and add 10 ml of H2O to each cup. 2. Before starting each test, pressure 10 ml of HOSTS in the HOSTS syringe, so the reaction can be stopped right away. 3. One person should add the reagents while another person keeps time. 4. For the 10 second time trial. With the transfer pipette, add 1 ml of catalane extract to the 10 sec cup. Right after hat, swirl the cup to mix the contents. The reaction will take place after adding catalane.
At 10 seconds, add the HOSTS to the cup. 5. Repeat with 30 sec, 60 sec, 120 sec, and 180 sec. Data/Results: Activity A results Testing for enzyme activity – The reaction was an enzymatic reaction because the chemicals and substances reacted. There was fizzing and bubbling after the reaction. Right when the chemical was added to the solution, the solution started fizzing right away. After about 5 seconds, small bubbles started forming, and they all moved towards the center of the cup. As a result, one big bubble formed hen they came together, and when the bubble got too big, it popped and the process would repeat over again.
The effect of boiling on enzyme activity – The reaction when the catalane was boiled, had the same results as the testing for enzyme activity results. The solution still boiled and formed bubbles, as well as the process of the small bubbles joining together. The only difference was that the reaction time was slower. This would repeat over and over slower than at room temperature. Testing for catalane in living tissue – For the last procedure of activity a, we had to find out whether living tissue would react to catalane, such s a potato. We mashed up the potato with the glass rod, and we added H2O for the reaction.
According to observations, the only thing that happened was that bubbles formed and the potato got soggy after 24 hours in the catalane. Activity B Initial Volume Final Volume 1 ml A Volume 4 ml In activity b, we composed a baseline that held H2O, H2O, and 1 M HOSTS. We swirled the contents to mix them together. With the 5 ml syringe labeled transfer, we removed 5 ml of the baseline solution put it in the cup labeled titration. After that, we titrated the solution to determine the baseline amount of H2O. The solution turned a brown color when we added Km to titrate it.
The initial volume was 5 ml, the final volume is 1 ml, and the change is 4 ml. Activity c 5 ml 0. 6 ml 4. 4 ml In activity c, we made unconcealed decomposition with H2O, 20th, and 1 M HOSTS. During this part, we had to take the 5 ml syringe labeled transfer and remove 5 ml of the mixture and put it in the titration cup. We titrated the sample by adding 5 ml of Km to determine the amount of H2O left in the solution after 24 hours. The titration turned brown and over a 24 hour time period it remained the same color when exposed to oxygen. Activity D 10 sec 30 sec 60 sec 120 sec sec 3. 1 ml 6 ml 2 ml 18 ml 2. 8 ml 2. Ml 3. 4 ml 3 ml 3. 2 ml 2. 2 ml . 2 ml 2. 6 ml H2O decomposed 0. 018 ml 0. 2 ml 0. 81 ml 2. 81 ml . 41 ml In activity d, we did time trials of how fast H2O decomposes to exposure of catalane. We gathered all the cups so we could have them ready for each time trial. For 10 seconds, we pre-filled the HOSTS syringe, so it would be ready right at 10 seconds. We added 1 ml of catalane to the 10 second cup, and above are the results. Lab Questions: 1 . Changes that I observed that indicates a chemical change was occurring were hat I observed fizzing and bubbles, so the solution is chemically changing due to a reaction. . The balanced equation for the reaction is E+S t] SEES 0 E+P 3. A. Substrate: S B. Enzyme: E C. Intermediate complex that formed: SEES D. Products: P 1. Procedure – 2. Boiling decreases the activity because Hydrogen bonds absorb excess heat and break the bonds. The enzyme will lose its structure and therefore its ability to bind to its substrate, this is called denomination. 3. Three factors that would affect the activity of catalane are temperature, pH, and concentration. Overall, all three of these will slow down catalane activity.
The temperature for catalane is 37 degrees Celsius, which his human body temperature, and changing the nature would just nature the catalane. Analysis of Results 1 . It is necessary to determine a baseline for H2O because the baseline is the controlled variable. Without the baseline, observations and recordings can’t be made. 2. The addition of HOSTS stops the reaction because HOSTS, or sulfuric acid, denatures the enzyme. Therefore the shape of the active site changes and it no longer matches the substrate, making it a different substance. 3. Pig. 5 | the dependent variable is the decomposed H2O and the independent arable is the times. 10 seconds, 30 seconds, 60 seconds, 120 seconds, and 180 seconds) 4. The line on my graph represents the data on table 3. It shows that at 10 seconds, . 018 ml of H2O was decomposed. For 30 seconds, . 20 ml was decomposed, etc. 5. The graph of table 3 shows the rate over time, and it is not fairly constant at all. From 60 to 1 20 seconds the decomposing rate was about 2 ml, but the 120 to 180 seconds decreases by 2. 6. On lab packet. 7. The initial rate of the catalane is approximately . 0649 ml/seconds. Conclusion: The enzyme catalyst lab’s purpose was to investigate enzyme reactions when exposed to different things.
In this lab, we put in catalane in a cup with H2O and observed, we boiled it, and we used live tissue, such as a potato. We also made mixtures and titrated them to see the amount of H2O in a solution after titration. Evermore, we did timed reactions to see the decomposing rate of the enzymes. In activity A, when we added the catalane to the H2O, bubbles started to form and there was slight fizz. The purpose of this lab is to see whether enzymatic reactions will take place on some substances and/or in certain states, such as boiling water and live potato tissue.
The bubbles got smaller, and they moved into the center. As for the boiling substance, it was the same, but the process was slower. I conclude that this occurred because the solution was changing state when it boils, so the particles move faster causing a slower reaction. In activity B and C we titrated the solutions, but they were slightly different. The point in this lab was to find out the baseline amount of H2O after titration. For activity B, we titrated a baseline consisting of H2O, HOSTS, and 20th. For activity C we did the same thing but we made unconcealed decomposition.