Year 11 Pre-Diploma Biology

ENZYME ACTIVITY AND TEMPERATURE

Temperature should affect the activity of enzymes in a predictable way, increasing their activity until
the enzyme (a protein) is denatured. At 0c we should expect a reaction which is catalysed by an
enzyme to be almost at a standstill. An interesting question which we can investigate is how fast does
the rate of a reaction (ie enzyme activity) increase as we raise the temperature. Many biological
reactions have a Q10 factor of 2, meaning that for every 10c increase in temperature, the speed of the
reaction doubles. Most proteins denature when the temperature reaches about 50c, which is when a
reaction would thus stop. Dont forget also the concept of limiting factors - a rate of a reaction may be
limited by other factors, such as the amount of enzyme present or the amount of substrate present.
All cells release the enzyme catalase in order to remove poisonous H 2O2, formed during respiration, as
quickly as possible. The H2O2 is converted into oxygen and water.
An important aspect of this investigation, which in itself is very simple, is to see if we can quantify
errors and uncertainties, and therefore give a degree of precision to the results.

Equipment, materials and chemicals

A potato
80+c
Scalpel
Cutting board
Balance with accuracy 0.1 gm
25 ml Hydrogen peroxide in a beaker

4 water baths at 0c, Room temp, 40c and

Pestle and mortar
10 ml measuring cylinder
5 test tubes

Procedure
1. Collect the apparatus, materials and H2O2. (The water baths should be ready for you and will be
shared by all groups.)
2. Remove the skin from a potato and cut about 10 gm into small pieces. Put these in the mortar.
3. With the pestle, grind up the potato in the mortar until it is a mush without big pieces.
4. Weigh out 4 EQUAL masses of the ground-up potato. An optimum mass is between 1.0 and 1.5
gm. Put one mass of potato in each of the test tubes.
5. Label the test-tubes 0, RT, 40 and 80.
6. When all 4 test tubes are ready, add 5 ml 10% H2O2 to each test tube
7. Quickly measure the height of the mixture in the test tube.
8. Quickly place the appropriate test tube in each of the water baths.
As the catalyse in the potato cells reacts with the H 2O2, foam will be given off which will rise up
inside the test tube.
8. Let the foam rise for a maximum of 10 minutes, and measure the new height of the foam. Record
these measurements in a table. Make any other observations which might be useful.
Results
Show the results graphically a very simple line graph would be sufficient. Your plot should indicate the
extent of uncertainties a whisker plot. Can you suggest a Q10 factor?
Conclusions and evaluation
Make a conclusion. This would be a reference of the results obtained, against a prediction.
Evaluate the procedure and the investigation, try to quantify the errors and uncertainties where
possible, and suggest ways in which the investigation might be improved if you were to repeat it.
John Osborne
October 2009