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Leader:
Rachel Yao

Members:
Janine Roselle de Vera

Camille Luis
Jason Aurigue
Kenneth Fontillas

Theoretical Background:
Experiment 7
Factors Affecting Enzyme Activity

Lunuelle Surriane Samonte

February 06, 2010
Group 8

A number of external factors affect the activities of enzymes in speeding the

conversion of reactants to products. These factors include variations in the
temperature, pH, concentration of enzyme, concentration of substrate molecules,
coenzymes and inhibitors.

Materials:

2% starch solution
0.1% a-amylase
0.5M sodium phosphate buffer pH 4.0
0.5M sodium phosphate buffer pH 7.0
0.5M sodium phosphate buffer pH 9.0
3M HCl solution

Methodology:

A. Effect of pH on Enzyme Activity

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B. Effect of Temperature on Enzyme Activity

Results:

A. Effect of pH on Enzyme Activity

Test tube

4
1
Buffer
Water

Sodium phosphate buffer

pH 4.0
Sodium phosphate buffer
pH 5.0
Sodium phosphate buffer
pH 7.0

B. Effect of Temperature on Enzyme Activity

Test tube Temperature of Water

Bath (°c)
20

37

60

80
Absorbance at 650mm
0.903

0.908

0.924

0.928

Absorbance at 650mm

0.982

1.070

0.947

0.945
Discussion:

After monitoring the experiment, we conclude that the temperature and pH of a

substance have a dramatic effect on the enzyme activity because as we know that
enzymes are sophisticated catalysts for biological processes. As they are flexible
polypeptide chains, their activity is affected by the chemistry of their surroundings and
by physical factors such as temperature and pH changes.

Question:

1. Write the chemical equation of the enzymatic reaction in the experiment. Which
species in the experiment contributes to the absorbance of the solution?

Na3PO4 + HCl = NaCl + H2PO4, Hydrochloric acid.

2. What is the effect of the pH of the solution to the enzymatic activity? How would
you explain the phenomenon?

Different pH results is observe (dramatically results) because each enzyme has an

optimal pH range that helps maintain its normal configuration in an environment in which it
operates.

3. What is the effect of temperature of the system to the enzyme activity? How
would you explain this phenomenon?

A higher temperature generally results in an increase in enzyme activity. As the

temperature rises, the movement of enzyme molecules and substrate molecules increases. This
causes more collisions between the enzyme and the substrate. If the temperature rises beyond
a certain point, however, the enzyme activity eventually levels out and then declines rapidly
because the enzyme is denatured by heat. The enzyme shape changes during denaturation and
cannot catalyze the reaction. Therefore there is a sharp drop in the rate of the reaction with the
change in temperature.

4. What is the purpose of adding 3M HCl in the solution?

Hydrochloric acid is a strong acid which means HCl dissociates or ionizes completely in
water. It serves as an acid-catalyst for chemical reactions where anhydrous (water-free)
conditions are desired.

Prepared by:
Janine Roselle P. de Vera