EXERCISE 8 : CHEMICAL AND PHYSICAL PROCESSES OF

DIGESTION
(Praktikum Fisiologi Modul Gastrointestinal)

Buat Laporan untuk Praktikum Fisiologi berikut dan jawablah

pertanyaan ini dalam Bahasa Indonesia

1. Activity 1 : Assesing Starch Digestion by Salivary Amylase

Experiment Data:

Tube Reagent Reagent Treatme Benedic

Reagent 3 Time Temp. IKI
No. 1 2 nt t's
pH 7.0
1 Amylase Starch Boiled 60 37 + -
Buffer
pH 7.0
2 Amylase Starch Frozen 60 37 - ++
Buffer
pH 7.0
3 Amylase Starch None 60 37 - ++
Buffer
Deionize
pH 7.0
4 Amylase d None 60 37 - -
Buffer
Water
Deionize
pH 7.0
5 d Starch None 60 37 + -
Buffer
Water
Deionize
pH 7.0
6 d Maltose None 60 37 - ++
Buffer
Water
pH 2.0
7 Amylase Starch None 60 37 + +
Buffer
pH 9.0
8 Amylase Starch None 60 37 + +
Buffer

Post Lab Quiz

1. Tubes 3,7, and 8 reveal that ...
a. Amylase activity was unaffected by pH
b. Amylase activity was highest at pH 9
c. Amylase activity was highest at pH 2
d. Amylase activity was highest at pH 7

2. This activity includes a number of negative controls. Which tube indicates

that the amylase solution was not contaminated with maltose?
a. Tube 1
b. Tube 4
c. Tube 6
d. Tube 7
3. Which tubes indicate that the deionized water did not contain contaminating
starch or maltose?
a. Tube 4 and 5
b. Tube 4 and 6
c. Tube 4,5 and 6
d. Tube 1,4 and 5
4. Explain where and why salivary amylase would be most active
a. Salivary amylase would be active in the mouth and the stomach because
pH 2 and pH 7 are where its peak activity occurs
b. Salivary amylase would be most active in the stomach because pH 2 is
where its peak activity occurs
c. Salivary amylase would be most active in the small intestine because pH
9 is where its peak activity occurs
d. Salivary amylase would be most active in the mouth because pH 7 is
where its peak activity is

Review Sheet
1. List the substrate and the subunit product of amylase!
2. What effect did boiling have on enzyme activity? Why?
3. At what pH was the amylase activity the most active? Describe the
significance of this result!
4. Briefly describe the need for controls and give an example used in this
activity!
5. Describe the significance of using a 37oC incubation temperature to test
salivary amylase activity!

2. Activity 2 : Exploring Amylase Substrate Specificity

Experiment Data:

Tube Reagent Benedic

Reagent 2 Reagent 3 Time Temp. IKI
No. 1 t's
pH 7.0
1 Amylase Starch 60 37 - ++
Buffer
pH 7.0
2 Amylase Glucose 60 37 - ++
Buffer
pH 7.0
3 Amylase Cellulose 60 37 + -
Buffer
Cellulos pH 7.0 Deionized
4 60 37 + -
e Buffer Water
Peptidas pH 7.0
5 Starch 60 37 + -
e Buffer
pH 7.0
6 Bacteria Cellulose 60 37 - ++
Buffer

Post Lab Quiz

1. The substrate(s) for amylase is/are..
a. Starch
b. Cellulose
c. Peptides
 d. Starch and cellulose
e. Starch and peptides

2. The result of tube 5 demonstrated that..

a. Peptidase can digest starch
b. Peptidase does not digest starch
c. Peptidase can digest cellulose
d. Peptidase can digest glucose

3. Why was the cellulose in tube 6 hydrolized to glucose?

a. The bacterial suspension contained the enzyme amylase, which digest the
cellulose
b. The bacterial suspension contained reducing sugars
c. The bacterial suspension contained the enzyme peptidase, which
digested the cellulose
d. The bacterial suspension contained the enzyme cellulase, which digested
the cellulose

4. Can you detect the presence of contaminating amylase from your

experiments?
a. There was no contaminating amylase because tube 4 was negative
b. There was contaminating amylase because tube 4 was positive
c. It is not possible to determine whether contaminating amylase is present
because amylase doesnt digest cellulose
d. There was no contaminating amylase because tube 4 was positive

Review Sheet
1. Describe why the result in tube 1 and tube 2 are the same!
2. Decsribe the result in tube 3!
3. Describe the usual substrate for peptidase!
4. Explain how bacteria can aid in digestion!

3. Activity 3 : Assesing Pepsin Digestion of Protein

Experiment Data:

Boile Optica
Tube d l
Reagent 1 Reagent 2 Reagent 3 Time Temp.
No. Densit
y
pH 2.0 +
1 Pepsin BAPNA 60 37 0.00
Buffer
pH 2.0 -
2 Pepsin BAPNA 60 37 0.40
Buffer
Deionized pH 2.0 -
3 Pepsin 60 37 0.00
Water Buffer
Deionized pH 2.0 -
4 BAPNA 60 37 0.00
Water Buffer
pH 7.0 -
5 Pepsin BAPNA 60 37 0.03
Buffer
pH 9.0 -
6 Pepsin BAPNA 60 37 0.00
Buffer
 Post Lab Quiz
1. Pepsin would be most active
a. In the mouth
b. In the stomach
c. In the small intestine
d. In the mouth and in the stomach

2. Which two tubes validated the result of the experiment?

a. Tubes 1 and 2
b. Tubes 2 and 3
c. Tubes 3 and 4
d. Tubes 4 and 5

3. With more enzyme activity the optical density ?

a. Increased
b. Decreased
c. Stayed the same

4. If pepsin were digesting an actual protein substrate, the product would be

a. peptides
b. amino acids
c. Peptides and amino acids
d. BAPNA

Review Sheet
1. Describe the effect that boiling had on pepsin and how could you tell it had
that effect
2. At which pH do you think pepsin will have the highest activity? Was your
prediction correct about the optimal pH for pepsin activity?
3. What do you think would happen if you reduce the incubation time to 30
minutes for tube 5?

4. Activity 4 : Assesing Lipase Digestion of Fat

Experiment Data:

Tube
Reagent 1 Reagent 2 Reagent 3 Reagent 4 Time Temp. pH
No.
pH 7.0
1 Lipase Vegetable Oil Bile Salts 60 37 6.21
Buffer
Deionized pH 7.0
2 Lipase Vegetable Oil 60 37 7.00
Water Buffer
Deionized pH 9.0
3 Lipase Bile Salts 60 37 9.00
Water Buffer
Deionized pH 7.0
4 Vegetable Oil Bile Salts 60 37 7.00
Water Buffer
pH 2.0
5 Lipase Vegetable Oil Bile Salts 60 37 2.00
Buffer
6 Lipase Vegetable Oil Bile Salts pH 9.0 60 37 9.00
 Buffer

Post Lab Quiz

1. What is the product of lipase hydrolysis?
a. Bile salts
b. Triglycerides
c. Phospholipids
d. Fatty acids

2. From your results, which pH is ideal for pancreatic lipase digestion?

a. pH 2.0
b. pH 7.0
c. pH 9.0

3. Which tube confirms that there is no lipase in bile salts or vegetable oil?
a. Tube 2
b. Tube 3
c. Tube 4
d. Tube 5

4. From your results, where (in theory) would pancreatic lipase be active ?
a. mouth
b. stomach
c. pancreas
d. mouth and pancreas

Review Sheet
1. Explain why you cant fully test the lipase activity in tube 5!
2. Which tube had the highest lipase activity? How well did the results
compare with your prediction? Discuss possible reasons why it may or may
not have matched !
3. Explain why pancreatic lipase would be active in both the mouth and the
pancreas !
4. Describe the process of bile emulsification of lipids and how it improves
lipase activity !

GOOD LUCK !!