B.

DIGESTION IN THE STOMACH

1. Peptic Digestion:

OBJECTIVES:
1. To determine the optimum conditions for the digestive enzymes;  2. To investigate the activity of these digestive enzymes  3. To illustrate their efficiency; and  4. To perform methods that will determine enzymatic activity

INTRODUCTION:

METHODOLOGY:

¼ cu. 0. 0.cm) .5 N ml ml ml ml 1 5 _ _ _ _ * 2 _ 5 _ _ _ * 3 4 _ 1 _ _ * Hard-boiled egg white* (*equal slices. 4 N Na2CO3.         Test tube Number Neutral Pepsin. 2% ml HCL. 2. SHAKE EACH TUBE AND DETERMINE THE PH USING PH PAPER.1.01 N HCL.05 N Lactic Acid. 0. PREPARE TEST TUBES ACCORDING TO THE PLAN PRESENTED.

5 N ml Hard-boiled egg white* (*equal slices. 0. 2% ml HCL. 0.        Test tube Number Neutral Pepsin.01 N ml HCL. 4 N ml Na2CO3.05 N ml Lactic Acid. 0.cm) 4 4 _ _ 1 _ * 5 4 _ _ _ 1 * . ¼ cu.

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Test tube Number 1 5) (Neutral Pepsin. 2% ml Slightly Acidic .

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Test tube Number 2 (HCL. 0.01 N ml 5) Acidic .

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0.05 N ml 1) Acidic .Test tube Number 3 (HCL.

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Test  tube Number 4 Acidic (Neutral Pepsin. 2% ml 4)  (Lactic Acid. 4 N ml 1) .

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Test  tube Number 5 1) (Neutral Pepsin. 0.5 N ml Basic . 2% ml 4)  (Na2CO3.

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 3.  5.  4. Cover and incubate in your locker till the next laboratory meeting.  Determine the extent of digestion visually by noting the size of the hardboiled egg white. Add 4 drops of toluene. . Avoid shaking the tubes after the addition of the hard-boiled egg white. Add equal slices of hard-boiled egg white to each tube.

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TEST TUBE NUMBER  1 NO REACTION  NO VISIBLE CHANGE IN SIZE .

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 SLIGHT ENLARGEMENT OF THE HARD BOILED EGG WHITE .TEST TUBE NUMBER  2 THERE IS VISIBLE CHANGE IN SIZE.

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TEST TUBE NUMBER 3 THERE IS VISIBLE CHANGE IN SIZE.  SLIGHT ENLARGEMENT OF THE HARD BOILED EGG WHITE  .

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 SLIGHT ENLARGEMENT OF THE HARD BOILED EGG WHITE  .TEST TUBE NUMBER 4 THERE IS VISIBLE CHANGE IN SIZE.

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TEST TUBE NUMBER 5 NO REACTION  NO VISIBLE CHANGE IN SIZE  .

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2.  To each 1 ml supernatant liquid. .   7. Mix well and observe any resulting color. 3 and 4 with solid NA2CO3 first. Determine the extent of digestion by means of the Biuret test as follows. let stand and withdraw 1 ml of the supernatant liquid.6. add 1 ml of 10% NaOH and 2 drops of 1% CuSO4.  Neutralize those from the test tubes no.  Shake the tubes.

RESULTS: .

•TEST TUBE NUMBER 1 .

TEST TUBE NUMBER 2 .

TEST TUBE NUMBER 3 .

TEST TUBE NUMBER 4 .

TEST TUBE NUMBER 5 .

 (+) test = purple (presence of proteins)  (−) test = blue to pink when combined with shortchain polypeptides (it will not cleave on all peptide chains)  .BIURET TEST: A chemical test used to detect the presence of peptide bonds.

5%peptone (Pinkish solution). Compare colors obtained in each test tube with that given by 0. NO AVAILABLE 0. If there is no digestion yet. incubate longer.5% PEPTONE  .

5 to 2). its primary site of synthesis and activity is the stomach (pH 1. WHAT ARE THE OPTIMUM CONDITIONS FOR THE ACTIVITY OF PEPSIN? RENNIN? Pepsin is most active in acidic environments between 37°C and 42°C.QUESTIONS: 1. Pepsin exhibits maximal activity at pH 2.  .Accordingly.0.

. as this closely resembles the acidic environment of the stomach where rennin is most commonly found. although at a much lower level of reaction.5. Optimal pH for rennin is around 6. This is necessary because sometimes the stomach is diluted by outside materials.0-6. However rennin also works at neutral pH.

this enzyme is said to be absent in the stomach of adults. On the other hand. In this case. Calcium paracasenate is necessary in the process.  Infant curdling is catalyzed by the enzyme rennin acting on the hydrolysis of milk protein “Casein”. DIFFERENTIATE INFANT CURDLING FROM ADULT CURDLING. . curdling is catalyzed by hydrochloric acid and pepsin.2.

5–2. At pH 5. .0. In the intestine the gastric acids are neutralized (pH 7). WHY IS THERE NO DIGESTION OF PROTEINS IN GASTRIC JUICE WITH PH ABOVE 7?  The digestive power of pepsin is greatest at the acidity of normal gastric juice (pH 1.5).5 and above. and pepsin is no longer effective.0 activity begins to decline as the pepsin protein denatures and is inactive at pH 6. however pepsin is not fully denatured or irreversibly inactivated until pH 8.3.

4. Aside from. WHAT SUBSTANCES OTHER THAN HYDROCHLORIC ACID CONTRIBUTE TO THE ACIDITY OF GASTRIC CONTENTS?  Hydrogen ions contribute to the acidity of the gastric contents. There are no exact proportions between the different acid types. resulting in the pH fluctuation. Lactic acid produced by fermentation is another contributing factor. . the stomach also consists of large quantities of potassium chloride (KCl) and sodium chloride (NaCl).

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