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54 PhysioEx™ Exercise 8
Activity 2: Assessing Cellulose Digestion
Tubes #4, 5, and 6 showed that starch or cellulose was still present.
Tubes #1, 2, 3, and 7 showed positive tests for the Benedict’s reagent, indicating the pres-
ence of reducing sugar.
Freezing had no effect.
Freezing does not restrict enzyme activity, unlike boiling.
Amylase had no effect on the cellulose in tube #4.
Cellulose is digestible by bacteria.
Peptidase had no effect on animal starch. Peptidase does not work on carbohydrate sub-
strates so has no effect on digestion of these molecules.
Activity 4: Assessing Fat Digestion by Pancreatic Lipase and the Action of Bile
7. Tube 1 investigated the action of bile on enzyme activity, and tube 2 examined lipase
activity without bile. Bile enhances fat digestion by lipase.
Yes, you can determine if activity occurred in tube 6 because the pH would drop below
pH 9.0. A small amount of fat digestion occurred because pH decreased from 9.0 to 8.97.
The optimim pH for lipase activity was pH 7.0.
Using a pH method to assay for activity at pH 2.0 does not work because the buffer is
already quite acidic. There could have been activity at pH 2.0 that was not detectable by
this method.
In theory, lipase would be active in the mouth because its pH optimum is relatively neu-
tral. However, it would not be active in the stomach because of the acidic pH condition.
The substrate is vegetable oil (fat). The subunit formed is fatty acid (and monoglycerides).
PhysioEx™ Exercise 8 55
8
R E V I E W S H E E T
EXERCISE
NAME ____________________________________
Chemical and Physical
LAB TIME/DATE _______________________
Processes of Digestion
Carbohydrate Digestion
The following questions refer to Activity 1: Assessing Starch Digestion by Salivary Amylase.
1. At what pH did you see the highest activity of salivary amylase? 7 Why?
Salivary amylase is active in the mouth, which has a pH of 7.
2. How do you know that the amylase did not have any contaminating maltose?
3. What effect did boiling have on enzyme activity? Why? Boiling caused the protein salivary amylase to be denatured, thus
4. Describe the substrate and the subunit product of amylase. The substrate is starch; the product is maltose.
5. Does amylase use cellulose as a substrate? Explain. No; amylase is an enzyme that does not digest cellulose, only starch.
6. Did freezing have an effect on the activity of amylase? Explain. No; the enzyme activity was the same at freezing as it was at 37°.
7. Do you think that the bacterial suspension contained the enzyme cellulase (an enzyme that digests cellulose)? Why or why not?
8. What is the substrate of peptidase? Explain, based upon your results. In this simulation, BAPNA was the substrate of peptidase
because the optical density (indicating enzyme activity) increased in the solutions containing both BAPNA and peptidase.
57
Protein Digestion by Pepsin
The following questions refer to Activity 3: Assessing Protein Digestion by Pepsin.
9. At which pH did you see the highest activity of pepsin? 2 How does this correlate to the location of pepsin in the
10. What effect did boiling have on pepsin? Pepsin did not digest BAPNA in tube 1 so it can be concluded that boiling denatures pepsin.
11. Was there any digested BAPNA contaminating the pepsin or deionized (DI) water? No
How can you tell? Tubes 3 and 4 had an optical density reading of 0.0.
What is the usual substrate for pepsin, and what subunits are formed with pepsin activity?
Substrate—proteins
Subunits formed—proteoses, peptones, peptides (all small protein fragments), and free amino acids
13. What was the effect of decreasing the incubation time on the optical density results?
14. What effect would decreased incubation temperature have on pepsin activity? Why?
Decreased incubation temperature reduced pepsin activity because the optimal temperature for pepsin activity is body temperature.
15. What was the significance of using 37°C for the incubation? 37°C is body temperature.
16. Describe the activity of lipase with and without the addition of bile salts. Refer to Chart 4 for pH values. The activity of
17. Is the activity of bile a chemical or a physical process? Explain. Bile activity is a physical process; it breaks fat particles into
58 Review Sheet 8
18. What pH resulted in the maximum pancreatic lipase activity? 7.0
How does this optimal pH correlate to the enzyme’s location in the body? The pH of the small intestines is neutral.
19. Explain whether or not we can determine fat hydrolysis in tube 5. Why or why not?
We cannot determine if fat hydrolysis occurred in tube 5, which had a buffer of 2.0, and the pH of the solution after incubation was still
2.0. There could have been activity at pH 2.0 that was not detectable by this assay method.
21. Explain the significance of the movement of the tongue during swallowing. The tongue initiates and controls the buccal
22. Describe three events that occur during the pharyngeal-esophageal phase of deglutition. Mouth, nasopharynx, and larynx
are blocked; upper esophageal sphincter relaxes to open esophagus; food moves through esophagus by pressure gradients created
by peristalsis.
23. What was the time interval that you recorded between the first and second sound?
1–2 seconds
Review Sheet 8 59