The effect of temperature on the reaction time of the

enzyme peroxidase
By Denae Pitt-Nynde Wright
What are enzymes?
Enzymes are special types of proteins. Like all proteins, enzymes are
made from strings of amino acids. The function of the enzyme is
determined by the sequence of amino acids, types of amino acids,
and the shape of the string.
What do enzymes do?
Enzymes are responsible for a lot of the work that is going on in
cells. They act as catalysts in order to help produce and speed up
chemical reactions. When a cell needs to get something done, it
almost always uses an enzyme to speed things along.
Enzymes are Specific
Enzymes are very specific. This means that each type of enzyme
only reacts with the specific type of substance that it was made for.
This is important so enzymes don't go around doing the wrong thing
and causing chemical reactions where they are aren't supposed to.
How Enzymes Work
Enzymes have a special pocket on their surface called an "active
site." The molecule that they are supposed to react with fits neatly
right into that pocket. The molecule or substance that the enzyme
reacts with is called the "substrate."
The reaction takes place between the enzyme and the substrate at
the active site. After the reaction is complete, the enzyme releases
the new molecule or substance. This new substance is called the
"product."

Background: Hydrogen peroxidase (catalase) is an enzyme found

in many cells. This enzyme catalyses the breakdown of toxic
peroxide into useful oxygen gas and liquid water.

2H2O2(1) (Hydrogen peroxide) O2(g) (Oxygen) + 2h2O(1)

(Water)
Purpose: To determine the relationship between temperature and
enzyme activity.
Hypothesis: Out of the 3 temperatures of water, room temperature
will show the most amounts of oxygen bubbles due to the fact that
room temperature is the most comfortable temperature for
enzymes.
Variables: I.V: Temperature of the water
D.V: The amount of oxygen bubbles as a result of adding hydrogen
peroxide solution.
Apparatus:
6x Identical test tubes
Dropper bottle of H2o2
Distilled water bottle
Scalpel
Thermometer
4x 250ml beakers
Pasteur pipette
Ruler
Forceps
Ice cubes
Stopwatch
Test tube rack
Boring tool
8x medium test tubes
Hot water
Procedure:
Part A: Preparing hydrogen peroxide solutions
Use a 10ml-measuring cylinder to prepare 6 hydrogen peroxide
solutions as follows:
1. Add twenty drops of 6% hydrogen peroxide solution to a 10ml
measuring cylinder
2. Make up the volume in the measuring cylinder to 5ml with
distilled water using a Pasteur pipette, each time
3. Pour the contents of the measuring cylinder into a test tube
4. Repeat steps 1 to 3 five times
Part B: Varying Substrate Temperature
5. Add crushed ice to a beaker to a depth of 5cm
6. Add tap water to a beaker to a depth of 5cm

7. Add hot water to a beaker to a depth of 5cm

8. Add boiling water to a beaker to depth of 5cm
9. Place a thermometer into the water of each water bath
10.
Place a test tubes of diluted hydrogen hydroxide
solution into each beaker
Part C: Preparing potato cores
11.
Prepare a number of potato cores using the boring tool.
12.
Cut the potato cores into 2cm lengths, making sure each
core has identical dimensions.
Part D: Varying Enzyme Temperature
13.
Place one of the potato cores into each of the three
empty test tubes
14.
place one of these test tubes into each of the water
baths, next to the substrate test tube.
15.
Allow the five minutes for the test tube contents to
equilibrate with its surroundings.
Part E: Reaction
16.
Record the temperature of the ice water bath
17.
Pour the contents of the hydrogen peroxide tube from
the ice water bath into the tube containing the potato core
from the same water bath
18.
Place this test tube back into the ice water bath, and
allow the reaction to progress for 5 minutes
19.
Measure the height of the column of oxygen bubbles in
the test tube in mm
20.
Repeat steps 16 to 19 for the tubes in the other water
baths.
Results:
Beaker
#1
#2
#3

Water Temperature
Cold 6C
Room temperature
17C
Hot 58C

Height of oxgygen
bubbles
25 mm
25 mm
5 mm

Discussion:
Questions:
1. State the name of the enzyme used in this experiment
and what its substrate was. What was the source
(where it came from) of the enzyme?
Hydrogen peroxidase was the enzyme used and the substrate
used was hydrogen peroxide.
2. State all possible experimental errors
Although the beakers with the ice cold and boiling hot water
werent left out in room temperature for long, they were still
left out consequently increasing/decreasing the temperature
of the water therefore altering the final result.

3. State 2 experimental improvements

1. Prepare potato core before pouring the water into the
beakers so the water is as hot/cold as possible.
2. Organize tools before starting the experiment so less time
is wasted searching and preparing.

4. Why was room temperature the best environment for

this particular enzyme?
If the temperature of the system is increased slightly, the
substrates will have more energy and will therefore collide
more frequently, allowing for more reactions. However, if the
temperature increases significantly, such as to the boiling
point of water, they become extremely deformed, and since
their function was dependent on their shape, this cripples
their ability to catalyze reactions. While a few bonding sites
may still be active, many will not, and those that are will have
lost some of their functionality due to the change in shape
Conclusion:
Out
of
the 3

temperatures of
water, room
display the
of bubbles as
hypothesis,
the cold
surprising

temperature did
highest amount
stated in the
although, so did
water which was
because it was
presumed that cold
water would
produce the least
amount
of bubbles as it mightve been the least desired site for the
hydrogen peroxide to set off a reaction

The experiment was a success in more ways than not. We collected

information regarding what peroxide does and what hydrogen is,
however, these results may not have been as accurate as possible
due to letting the hot and ice cold water sit out in room temperature
for a little over 5 minutes before applying the potato core. Overall,
the individuals in the group conducted the experiment to the best of
their abilities and though there were some minor errors the
experiment was still a success.