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10 views

jBvNdGgbK7

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akkuy975
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Study of Digestion of Starch by Salivary Amylase and Effect of pH and Temperature on It

Objective

To study the digestion of starch by salivary amylase and observe the effect of pH and temperature on the enzymatic activity.

Introduction

Enzymes are biological catalysts that accelerate chemical reactions in living organisms. Salivary amylase, an enzyme present in saliva, plays a
key role in the digestion of carbohydrates by breaking down starch into maltose. The activity of salivary amylase is influenced by factors such
as pH and temperature. Understanding these effects provides insights into enzymatic behavior and digestion processes.

Requirements

1. Freshly prepared saliva sample


2. Starch solution (1%)
3. Iodine solution
4. Test tubes
5. pH buffer solutions (pH 4, pH 7, pH 9)
6. Thermometer
7. Water bath
8. Beaker
9. Pipettes and droppers
10. Stopwatch

Theory

Salivary amylase catalyzes the hydrolysis of starch into maltose. The reaction is as follows:

Starch+Water AmylaseMaltose\text{Starch} + \text{Water} \xrightarrow{\text{Amylase}} \text{Maltose}

The activity of salivary amylase is optimal at a specific temperature (around 37°C) and pH (near neutral). Extreme variations in pH and
temperature can denature the enzyme, reducing its efficiency.

Procedure

1. Preparation of Materials
Collect a fresh saliva sample and dilute it with distilled water (1:10 dilution).
Prepare a 1% starch solution by dissolving 1 g of starch in 100 mL of water and boiling it until fully dissolved. Cool the solution before use.
2. Effect of pH on Amylase Activity
1. Label three test tubes as pH 4, pH 7, and pH 9.
2. Add 5 mL of starch solution to each test tube.
3. Add 1 mL of the respective pH buffer to each test tube.
4. Add 1 mL of the diluted saliva sample to each tube and mix thoroughly.
5. Place the test tubes in a water bath maintained at 37°C.
6. At intervals of 1 minute, take a small sample from each test tube and mix it with a drop of iodine solution on a white tile.
7. Record the time taken for the blue-black color (indicative of starch) to disappear, showing complete digestion.

3. Effect of Temperature on Amylase Activity


1. Label four test tubes as 0°C, 25°C, 37°C, and 60°C.
2. Add 5 mL of starch solution and 1 mL of pH 7 buffer to each tube.
3. Place the tubes in water baths set to the respective temperatures for 5 minutes to equilibrate.
4. Add 1 mL of diluted saliva sample to each tube and mix thoroughly.
5. At intervals of 1 minute, test the reaction mixture with iodine solution and record the time taken for the blue-black color to disappear.

Observations

pH Time for Complete Digestion (Minutes)


4
7
9
Temperature (°C) Time for Complete Digestion (Minutes)
0
25
37
60

Results

1. The enzymatic activity of salivary amylase was highest at pH 7 and 37°C.


2. At pH levels far from neutral, the enzyme showed reduced activity.
3. Extreme temperatures (very low or very high) negatively affected the enzyme’s ability to digest starch.

Conclusion

Salivary amylase effectively digests starch under optimal conditions of pH (around 7) and temperature (around 37°C). Deviations from these
conditions result in decreased enzymatic activity due to denaturation or reduced efficiency.

Precautions
1. Use freshly prepared solutions to ensure accuracy.
2. Maintain the specified temperatures accurately during the experiment.
3. Rinse all apparatus thoroughly to prevent contamination.
4. Handle iodine solution with care as it can stain.

References

1. NCERT Biology Textbook for Class XII.


2. Laboratory Manual for Biology, Class XII.
3. Online resources on enzymatic activity and digestion processes.

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