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Title:

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.