The Effect of Substrate Concentration and pH on Enzyme-Controlled Reactions Lab
Purpose: We had to find out what effect a given amount of substrate concentration and different ph levels had on enzyme-controlled reactions.
Hypothesis: If I add more substrate concentration, then there will be more of a reaction because the higher amount of substrate concentration, the bigger the reaction.
If I make the pH higher, then there will be more of a reaction because the higher the pH the bigger the reaction will turnout.
Materials: A computer Paper (to record results) Pencil (to record results)
Procedures for substrate concentration experiment: 1. Go to the website http://glencoe.mcgraw- hill.com/sites/dl/free/0078695104/383930/BL_11.html Remember: The pH level is adjusted by clicking the up or down arrows below the corresponding test tube To add substrate, click and drag the weighing paper to the appropriate test tube for all trials, assume the enzyme concentration in each test tube is constant (the same for each) 2. Adjust the pH level of each test tube to 7 3. Add .5 g substrate to Test Tube 1, 1.0 g substrate to Test Tube 2, 2.0 g substrate to Test Tube 3, 4.0 g substrate to Test Tube 4, 8.0 g substrate to Test Tube 5 4. Click on the computer screen to observe the calculated data. Record the data (Number of Molecules of Product) in Data Table 1: Substrate Concentration in the column pH 7 5. Click the Reset button 6. Adjust the pH level of each tube to 3 7. Repeat steps 2 and 3, making sure to record all data in the appropriate column 8. Click the Reset button and repeat the experiment one more time after adjusting each test tubes to a pH level of 11
Procedures for pH level experiment: 1. Go to the website http://glencoe.mcgraw- hill.com/sites/dl/free/0078695104/383930/BL_11.html 2. Adjust the pH level of test Tube 1 to 3, Test Tube 2 to 5, Test Tube 3 to 7, Test Tube 4 to 9, and Test Tube 5 to 11. The test tubes should be reset to these pH values for each trial outlined below 3. Add 0.5 g substrate to each of the test tubes 4. click on the computer screen to observe the calculated data. record the data in Data Table 2: pH Level in the column 0.5 g 5. Click the Reset button 6. Adjust the pH levels as stated in step 1 7. Add 2.0 g substrate to each of the test tubes 8. click on the computer screen and record your data in the appropriate column 9. click the Reset button 10. repeat steps 1-3 using 8.0 g substrate
Observation/Data: Data Table 1: Substrate Concentration For Test Tube 1, I added 0.5 g of substrate and the number of molecules of product formed per minute for pH 7 was 72, for pH 3 19, and for pH 11 24. For Test Tube 2, 1.0 g of substrate was added. Set at pH 7, the number of molecules of product formed per minute was 145. At pH 3, it changed to 39, while pH 11 gave the result of 49. Test Tube 3 gave different results, as pH 7 gave 300, pH 3 gave 82, and pH 11 gave 103. The results for Test Tubes 4 and 5, the results were the same with pH 7 giving the results of 350. pH 3 being 96 and pH 11 turning out to be 121. This data shows that for each pH, the number of molecules of product formed per minute increased, unsteadily, and the leveled off after 4.0 g of substrate was added. The variables were the pH level and the number of molecules of product formed per minute, and the control was the amount of substrate.
Data Table 2: pH Level For Test Tube 1, the pH level was set at 3, Test Tube 2s pH level at 5, Test Tube 3s pH level at 7, Test Tube 4s pH level at 9, Test Tube 5s pH level at 11, and with the total number of molecules of product formed per minute resulting to be as follows: Test Tube 1 at 0.5 g of substrate being 19, 2.0 g gave results ended up being 82, and at 8.0 g, 96; Test Tube 2 at 0.5 g resulted to be 39, while 2.0 g of substrate ws 168, and 8.0 g of substrate totaled 198; following was Test Tube 3 at 0.5 g of substrate was 72, 2.0 g of substrate being 300, and 8.0 g of substrate ended up being 350; next was Test Tube 4 at 0.5 g of substrate resulted as 45, while 2.0 g of substrate ended at a total of 189, and 8.0 g of substrate being 223; and lastly Test Tube 5 at 0.5 g of substrate ended as a total of 24, 2.0 g of substrate at 103, and 8.0 g of substrate finishing at 121. The variables were the amount of substrate added to each test tube and the number of molecules of product formed per minute, while the control was the pH level.
Conclusion: The purpose of this experiment was to see, first, how substrate concentration affected enzyme controlled reaction, and how pH affects enzyme controlled reactions. The conclusion was that the amount of substrate, x, would make the enzyme controlled reaction bigger up to a certain point (4.0 g). Another conclusion was that different pH levels would make the reaction peak (at pH 7) and then decrease. I think this was a fair experiment because everything was constant. Nothing could have changed because this was a computer lab. The things used were to be assumed as constant. In real life, these things would be most likely not be as steady. I learned that pH levels and different amounts of substrate go hand in hand. Their results depend on each other. Some good questions for further experimentation could be what would the effect be if amounts of substrate higher than 8.0 g were placed in different levels of pH.