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Running head: EFFECT OF pH LEVEL ON ENZYME ACTIVITY 0

LOW pH AND ENZYME ACTIVITY

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Introduction:

Within a cell, enzymes are used as catalyst to increase the rate of chemical reaction. They do not consume themselves but just help in increasing the rate of reaction. Within a body, enzymes are of different types depending upon their specific functions. Hydrogen peroxide is a toxic chemical. It breaks down into harmless oxygen and water. This reaction can be speed up using the enzyme catalyst produced by yeast. Hydrogen peroxide is produced as byproduct in cellular reaction. As it is poisonous it must be broken down. Hence this reaction is important. The speeding up of reaction is shown below:

2H2O (aq) catalyses 2H2O (l) + O2 (g)

This experiment is designed to analyze how does the enzyme catalase activity gets affected by the pH levels. The experiment has also been designed to outline all the directions and the ways by which the observation can be made clearly and to accurately. Yeast, will be used as enzyme and hydrogen peroxide will be used as substrate. This experiment will be used to determine the effects of concentration of the hydrogen peroxide on the rate of reaction of enzyme catalase.

Hypothesis:

Presence of catalyst will cause hydrogen peroxide to break down into water and oxygen. The increase in pH of substrate will cause the increase in the rate of reaction. It will optimize at pH 7 and will diminish/ moderated after pH 9.

The important question here that will help in understanding the directions of the experiment are as below:

In what way does the rate of reaction depend on the pH of substrate?
At what level of the substrate concentration the reaction shows evidence of enzyme the saturation?

The variables

There will be independent and dependent variables:

Independent variable: In this experiment the independent variable are hydrogen peroxide and the amount of catalyses.

Dependent variables: In this experiment the dependent variable is pressure of the oxygen after catalyses reacts with peroxide.

The Experiment

The Materials used in the experiments are:

Beakers
Hydrogen peroxide
pH paper
5 clean test tubes
Thread and ruler or tape measure.
Vinegar
Balloons
Marker
Yeast suspension
Stop watch

The Procedure:

Clean and label 5 test tubes as 1, 2, 3, 4 and 5. Add 2 ml hydrogen peroxide in each of them. Now in test tube 1 add three drops of acetic acid available as vinegar. In test tube labeled as 2 add 5 drops of the vinegar from the vinegar jar. Same way, in test tube labeled 3, pour 7 drops of vinegar, in test tube labeled 4 pour 9 drops of the vinegar and in the test tube labeled 5 pour 11 drops of the vinegar.

Now with the use of pH paper, determine pH values of the each of the 5 solutions.

Now add the catalyst enzyme available as the yeast suspension in the test tube labeled 1 and then cover the tube immediately with a balloon as appropriate.

After this, observe the above reaction for ten minutes and then measure the circumference of the balloon used to cover the above solution.

For test tubes labeled 3 and 5, repeat the above mentioned steps.

Now fill the below table with the numbers obtained from the observation.

Test tube	pH level	Circumference of balloon (cm)

		11
		15
		12
	11	10

Qualitative observation

Test tube	pH Level	Observations
		On adding the yeast suspension to the hydrogen peroxide, there were bubbles seen and observed in the solution and the balloon increased by its volume. The solution changed its color and turned white.
		The solution in the test tube 2 bubbled and reached almost the top of the tube. There was more increase in the volume of the balloon as compared to test tube 1. The solution changed its color and turned white.
		The solution in the test tube 3 bubbled and reached the top of tube, almost spilling out in the balloon. The volume of the balloon did increased further. The Solution changed its color and turned white.
		The solution in test tube 4 bubbled and reached the top of the tube. The volume of the balloon did increase but not as much as it did in tube 3. The solution changed its color and turned white.
	11	The solution in test tube 5 produced bubbles, but they were little The volume of the balloon slightly increased The solution changed its color and turned white.

The Interpretation of observations:

The Enzyme used in the experiment is lactase. The Observations showed that it has the optimal level pH 7. Thus, oxygen was produced at high rate. Therefore, the inflammation of the balloon has a high volume. However, the lactase still functions between pH levels of 3 to 9. Any deviations in the pH outside of this range, specifically between the levels of 10 to 12, it would result to the inactivity of lactase as the lactase enzymes gets denatured at such high basic levels.

Conclusion:

In this experiment the observations that were made are consistent with the hypothesis made. During the experiment it was observed that the pH levels affect the activity of lactase enzyme. The enzymes denatures if the pH present is too high or too low. The pH 7 is optimal pH for lactase enzyme. At 7 the pH enzyme is the most active.

The Errors predicted in this experiment was the method that was used to conduct this experiment. The use of balloon in this experiment for measuring the volume of the oxygen gas which was produced was not that effective, as the pressure sensor for this purpose.

References:

Bennett, T. P., and Frieden, E., (1969). Modern Topics in Biochemistry, pg. 43-45, Macmillan, London

Harrow, B., and Mazur, A., (1958). Textbook of Biochemistry, 109, Saunders, Philadelphia

Holum, J.: Elements of General and Biological Chemistry, 2nd ed., 377, Wiley, NY (1968).

Martinek, R.: Practical Clinical Enzymology: J. Am. Med. Tech., 31, 162 (1969).

Pfeiffer, J.: Enzymes, the Physics and Chemistry of Life,pg 171-173, Simon and Schuster, NY (1954)