Experiment #: 3

TITLE: Enzymes

AIM: To determine the effect of pH on the action of the enzyme Catalase in chicken liver.

APPARATUS/MATERIALS: Petri dishes, beakers, dropper, hydrogen peroxide, distilled water, mortar, pestle,
test tubes, spatula, HCL, NaOH, balance, liver, Boiling tubes, Scalpel

PROCEDURE: Effect of pH:

1. three boiling tubes were labeled based on the different pH conditions that were investigated (neutral,
acidic, alkaline
2. three (3) drops of HCL were added to the test tube labelled acidic.
3. 2g of fresh liver was weighed
4. the liver was crushed using the mortar and pestle and then added to the tube containing HCL
5. 2cm3of hydrogen peroxide was added to the liver and HCL.
6. On a scale of 0-5 (0=no, 1=very slow, 2=slow, 3=fairly fast, 4=fast and 5=very fast) the rate of the
reaction was estimated based on how fast effervescence appeared in the tubes
7. The height of the bubbles in the tube were measured
8. steps 2-7 were repeated using NaOH (alkaline) and water (neutral condition)
9. Observations were then tabulated

BACKGROUND INFORMATION:

Enzymes are macromolecular biological catalysts. Enzymes accelerate chemical reactions. The molecules upon
which enzymes may act are called substrates and the enzyme converts the substrates into different molecules
known as products. Enzymes are proteins that act as catalysts for chemical reactions that take place in cells.
Sometimes reactions occur but they occur too slowly to be of benefit in living cells; catalysts speed them up so
they are useful. Catalysts play a role in the reaction but are not used up in the reaction. Enzymes are organic
catalysts which aid in facilitating chemical reactions in the body. Enzymes are needed for metabolic pathways
in the body, respiration, digestion and other important life processes. When enzymes function properly,
homeostasis is maintained. However, if an enzyme is lacking or has an incorrect shape due to a genetic
mutation, this can lead to disease within an organism. An example of this is the disease phenylketonuria or
PKU. Changes in pH may not only affect the shape of an enzyme but it may also change the shape or charge
properties of the substrate so that either the substrate cannot bind to the active site or it cannot undergo
catalysis. In general enzymes have a pH Optimum; however the optimum is not the same for each enzyme.
Initially, an increase in substrate concentration leads to an increase in the rate of an enzyme-catalyzed reaction.
... At higher temperatures, the protein is denatured, and the rate of the reaction dramatically decreases. An
enzyme has an optimum pH range in which it exhibits maximum activity.

OBSERVATIONS/RESULTS:

pH Observation Estimated rate of reaction Height of foam (cm)

condition (0-5 scale)
Neutral hot 5 16.5
Acidic warm 4 14.1
Alkaline cold 3 15.9
DISCUSSION:

In light of the outcomes acquired, it very well may be said that the Catalase had an optimum pH around 7.That
is on the grounds that the reaction was quickest when the pH was neutral in this way the chemicals would have
a situation appropriate to join with the individual substrate to form the products needed. Due to the substrate
meeting with the enzyme (at the active site) at such a fast rate the test tube felt hot. The second ph condition
was acidic and it had a rate of reaction of 4.Therefore the catalyst functioned admirably in this condition
however not in the same class as how they functioned in the neutral condition. Due to the substrate meeting
with the enzyme(at the active site) at such a fast rate the test tube felt warm. The alkaline condition had a
normal response time of 3.The catalysts worked at the slowest rate in this pH condition. Due to the substrate
meeting with the enzyme (at the active site) at such a slow rate the test tube felt cold.

SOURCES OF ERROR:

1. Ensure the scale is zeroed after the petri dish is placed on it.
2. Ensure liver piece are crushed properly.
3. Ensure liver pieces are crushed similar each time (because there will be great difference in surface area
to volume ratio which can also speed up the reaction).

CONCLUSION:

An experiment was done to determine the effect of pH on the action of the enzyme Catalase in chicken liver, the
results where as expected. In the alkaline solution the reaction was fairly fast, the acidic solution the reaction
was fast and finally in the neutral solution the reaction was very fast. The optimum pH is neutral as it is 7 which
is the pH of water.