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Cuyamaca College Enzymes pH and Lactase Physiology Lab Report

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1. In this activity, you will study the effect of pH, temperature, and lactose concentration on lactase
activity. By the end of this activity, you should be able to:
a. design experiments to study the effects of pH, temperature, and substrate
concentration on enzyme activity.
b. analyze data related to enzyme activity.
c. identify the components of the scientific method.
d. identify components of the substrate-enzyme-product system being studied.
e. summarize and communicate your lab data.

To prepare for the lab, watch the following video and think about enzymes, substrates,
products, maximum rate of reaction, and optimal conditions.


To complete the lab activity, review the Zoom link for lab posted on Canvas by Wednesday of
this week and go to the following website (https://sites.google.com/site/biologydarkow/lactaseenzyme-simulation The simulation is found on page 4 of the site or you can use the simulate
tab.

You do not submit this document on Canvas. This document includes the instructions you need
to complete the lab. You will submit the Lactase Enzyme Data document where you will report
your results.
Experiment I: The effect of pH on lactase activity. Click on the Simulate tab. The standard condition
settings are: lactose for 500 mg/dL , pH at 7.0 and temperature at 250C, and 5 mmol lactase enzyme
solution. You will vary one of these conditions in each experiment. In this Experiment I, you are varying
pH. The condition you are varying is the independent variable for that experiment. The outcome you
measure for that experiment is the dependent variable. The dependent variable in Experiment I is
glucose produced and is a measure of the rate of the enzyme activity. The hypothesis of an experiment
is a tentative proposed relationship of the independent and dependent variable of the experiment. The
hypothesis of Experiment I is pH does not affect the amount of glucose produced in a reaction catalyzed
by lactase.
Experiment I has two parts: I. determining the interval of interest and II.
determining the optimum pH for lactase activity.
I.
Determining the interval of interest. Before you can begin your experiment, you must
determine the interval of pH which will give you the most valuable, relevant data. Start by
running a simulation with the standard initial lactose, temperature and lactase enzyme
solution. Run simulations at a pH of 7. Record the glucose
concentration (mg/dL) at 4 (you do not need to record the data for 7 and 11 minutes in this
part) and repeat for a total of 5 runs. Repeat the simulations at a pH of 3, 5, 9, and
11. Make sure to have 5 runs for each pH setting. Record the glucose concentration at 4
minutes for each of the runs. Record all your data in the table.
pH
Run 1
(mg/dL)
Run 2
(mg/dL)
Run 3
(mg/dL)
Run 4
(mg/dL)
Run 5
(mg/dL)
Mean
(mg/dL)
Range
Range (High-Low)
3
5
7
9
11
Using the above data to produce a line graph (See the Zoom recording for this lab). Use the line graph to
help you determine the interval of interest. This will be covered in the Zoom lab.
II.
Use the data and graph to determine the pH interval which will give you the most valuable,
relevant data for determining the optimum pH. Divide this interval into ten (10) equally
spaced intervals (for example if you determine the interval between pH 7 and 11 is where
the peak enzyme rate is occurring, you will collect data at 7.4, 7.8, 8.2, 8.6 ….
10.2, 10.6 and 11). Adapt the data collection table to record the results of these simulations
at 4 minutes for five (5) runs for each pH level. Run the simulation and record the data.
pH
Run 1
(mg/dL)
Run 2
(mg/dL)
Run 3
(mg/dL)
Run 4
(mg/dL)
Run 5
(mg/dL)
Mean
(mg/dL)
Range
Range (High-Low)
Graph the data and determine the optimum pH (where maximum lactase enzyme activity is likely
located).
Experiment II: The effect of temperature on lactase activity. You will now use Experiment I as a model
to design an experiment to test the effect of temperature on lactase activity. Experiment II has two
parts: I. determining the interval of interest for the effect of temperature on lactase activity and II.
determining the optimum temperature for lactase activity.
First determine the independent variable of the temperature experiment. Next determine the
dependent variable of the temperature experiment. Identify the variables you will standardize in the
temperature experiment. Write out the hypothesis of your experiment. Record this information in the
Lactase Data Sheet.
I.
Determine the interval of interest for temperature. Choose a wide range of temperatures.
You can try a few temperatures to see those conditions result in any glucose production and
then work between the temperatures with low lactase activity. Determine these
temperatures and record them in the table below. Use the simulation website to determine
the glucose produced (rate of lactase activity) at 4 minutes for each of these temperatures.
Temp
0
C
Run 1
(mg/dL)
Run 2
(mg/dL)
Run 3
(mg/dL)
Run 4
(mg/dL)
Run 5
(mg/dL)
Mean
(mg/dL)
Range
Range (High-Low)
After you complete the table, use the data to determine the inflection point based on your results. The
interval of interest is the range of temperatures which will give you the most valuable, relevant data for
determining the optimum temperature in the second part of the experiment. Copy and paste the
above data table into the data document you will submit on Canvas.
II.
Use the temperature interval of interest to design the second part of the experiment.
Divide this interval into equally spaced intervals. Use the table below to record the results
of these five simulations at 4 minutes for each temperature. Run the simulation and record
the data.
Temp
0
C
Run 1
(mg/dL)
Run 2
(mg/dL)
Run 3
(mg/dL)
Run 4
(mg/dL)
Run 5
(mg/dL)
Mean
(mg/dL)
Range
Range (High-Low)
Use the data from the table to determine the optimum temperature (where maximum lactase enzyme
activity is likely located). Copy and paste the above data table into the data document you will submit
on Canvas.
Experiment III: The effect of lactose concentration on lactase activity. This experiment is NOT part of
the data sheet you will submit on Canvas. It is part of the quiz you will complete this week. For this
experiment, use Experiment I and Experiment II as models for studying the effect of lactose
concentration on lactase activity. Note that you will need to develop your own data collection tables to
record data. Design an experiment to test the effect of lactose concentration on lactase activity. What
is the independent variable of this experiment? What is the dependent variable of this experiment?
Which variables will you standardize? What is the hypothesis of this experiment? Describe your
experiments. What is the optimum lactose concentration for maximum lactase activity?
Experiment IV: Studying specificity using lactase activity. This experiment is part of the data sheet you
will submit on Canvas (question 5) and it is part of the quiz you will complete this week. Think about
the following question: Are you able to design an experiment to study specificity in lactase using the
Lactase Enzyme Simulation? Yes or No
Explain specificity and the experiment you would design to study this concept in lactase using the
simulation or how you would need to modify the simulation to test this concept. You don’t need to do
this experiment, just explain its design.
You do not submit this document on Canvas. This document includes the instructions you need to
complete the lab, but the data you submit is the completed Lactase Enzyme Data document.
Enzyme Lab Data
Answer the following questions, save this file as Word, Google doc, or PDF file, and submit on
Canvas before 1159PM Sunday. Be careful if you are using a Mac, as the default file extension
is page, not doc or docx. I cannot read .page files so pleas do not submit those files.
Review the module introduction including the video. Use Experiment II (Effect of
Temperature on Lactase Activity) results to complete 1-3. Use Experiment IV to complete 4.
Submit using Canvas prior to 11:59PM Sunday. You do not need to report information or results
from other Experiment I or III.
1. Write a hypothesis for Experiment II.
2. Identify the independent variable (IV) and dependent variable (DV) for Experiment II.
IV _________________________________ DV __________________________________
3.
Insert your completed data collection tables for Experiment II Part I and Part II below.
4. Complete the following summary table. Make sure your range data includes the absolute
difference between the high and low as well as the actual high and low for each of the
temperatures from Part II of Experiment II. Means and standard deviation must be
reported to the nearest 0.1 mg/dL.
Statistics Introduction Summary
Temperature (0C)
Mean (mg/dL glucose)
Range (mg/dL glucose)
SD (mg/dL glucose)
5. Experiment IV: Studying specificity using the lactase activity. Would you be able to
design an experiment to study specificity in lactase using the Lactase Enzyme Simulation
site? Yes or No Explain specificity and the experiment you would design to study this
concept in lactase using the simulation or how you would need to modify the simulation
to test this concept. You don’t need to do this experiment, just explain its design.
Enzyme Lab Data
Answer the following questions, save this file as Word, Google doc, or PDF file, and submit on
Canvas before 1159PM Sunday. Be careful if you are using a Mac, as the default file extension
is page, not doc or docx. I cannot read .page files so pleas do not submit those files.
Review the module introduction including the video. Use Experiment II (Effect of
Temperature on Lactase Activity) results to complete 1-3. Use Experiment IV to complete 4.
Submit using Canvas prior to 11:59PM Sunday. You do not need to report information or results
from other Experiment I or III.
1. Write a hypothesis for Experiment II.
Answer:
The temperature affects the amount of glucose produced in a reaction catalyzed by catalase.
2. Identify the independent variable (IV) and dependent variable (DV) for Experiment II.
IV _Temperature____________________ DV Amount of glucose produced______________
3.
Insert your completed data collection tables for Experiment II Part I and Part II below.
Answer:
Part I
Temp
0
C
Run 1
(mg/dL)
Run 2
(mg/dL)
Run 3
(mg/dL)
Run 4
(mg/dL)
Run 5
(mg/dL)
Mean
(mg/dL)
Range
Range (High-Low)
10
68.9
86.7
99.6
117
82.2
90.88
48.1 (117-68.9)
20
247
189
181
254
209
216
73 (254-181)
30
318
267
285
234
249
270.6
84 (318-234)
40
315
351
326
204
287
296.6
147 (351-204)
50
145
179
124
148
92.5
137.7
86.5 (179-92.5)
60
35.8
31.7
29.3
27.8
25.9
30.1
9.9 (35.8-25.9)
70
6.02
7.51
7.56
7.78
7.07
7.18
1.76 (7.78-6.02)
80
0
0
0
0
0
0
0 (0-0)
90
0
0
0
0
0
0
0 0-0)
100
0
0
0
0
0
0
0 (0-0)
Part II
Temp
0
C
Run 1
(mg/dL)
Run 2
(mg/dL)
Run 3
(mg/dL)
Run 4
(mg/dL)
Run 5
(mg/dL)
Mean
(mg/dL)
Range
Range (High-Low)
31
286
297
315
296
313
301.4
29 (315-286)
32
241
180
281
322
243
253.4
142 (322-180)
33
279
222
195
248
274
243.6
84 (279-195)
34
303
330
267
301
320
304.2
63 (330-267)
35
268
298
316
218
310
282
98 (316-218)
36
284
277
330
296
304
298.2
53 (330-277)
37
288
319
285
263
221
275.2
98 (319-221)
38
228
310
239
308
336
284.2
108 (336-228)
39
221
295
242
287
227
254.4
74 (295-221)
40
190
287
208
279
265
245.8
97 (287-190)
4. Complete the following summary table. Make sure your range data includes the absolute
difference between the high and low as well as the actual high and low for each of the
temperatures from Part II of Experiment II. Means and standard deviation must be
reported to the nearest 0.1 mg/dL.
Statistics Introduction Summary
Temperature (0C)
Mean (mg/dL glucose)
Range (mg/dL glucose)
SD (mg/dL glucose)
5. Experiment IV: Studying specificity using the lactase activity. Would you be able to
design an experiment to study specificity in lactase using the Lactase Enzyme Simulation
site? Yes or No Explain specificity and the experiment you would design to study this
concept in lactase using the simulation or how you would need to modify the simulation
to test this concept. You don’t need to do this experiment, just explain its design.

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