Aims:: Experiment: Rates of Enzyme Controlled Reactions

Experiment: Rates of Enzyme controlled reactions
Aims:
 To demonstrate some methods of studying rates of enzyme catalysed reactions.
 To look at the effects of pH, temperature and substrate concentration on the rate of
hydrogen peroxide breakdown in the presence of the enzyme catalase.
Introduction
One of the easiest methods of
measuring rates of reaction is to
measure the production of the products
of that reaction.
A common way to do this in reactions

which produce a gas is by collecting the
gas produced.
A simple inverted measuring cyllinder

filled with water over a water trough
works well.
Hypothesis
The breakdown of hydrogen peroxide into water and oxygen will happen more quickly when:
 the concentration of substrate is higher

 the pH is closest to the optimum pH of the catalase (near pH7.0 in humans)
 the temperature is closest to the optimum temp. (near 37°C in humans)
Apparatus
 A source of catalase; Equation of the breakdown
Safety
powdered yeast
 5 cm3 syringe / pipette Catalase Wear eye protection
 Dropping pipette
 25ml Measuring cylinder Hydrogen peroxide  Water + Oxygen
Tie back long hair
 Water bath set to 80°C 2H2O2 2H2O O2
 Stopwatch
 Solutions of Hydrogen reactant products
peroxide (20Vol, 15Vol,
10Vol, 5Vol, and 0Vol)
 Access to an electronic balance
 Access to buffers of pH 2,4,6,8,10
 Access to water baths of 10, 30, 50, 70, and 90 °C
© David Faure, InThinking www.biology-inthinking.co.uk

1
Method A: Measuring the Rate of the Reaction

a) Measure 0.1g of yeast powder using an electronic balance. The yeast cells contain the
enzyme catalase.
b) Measure 5 cm3 of 20 volume hydrogen peroxide into
a boiling tube.
Safety
c) Set up the inverted measuring cylinder as in the
Wear eye protection.
diagram above.
d) Add the yeast to the boiling tube and quickly
Rinse any splashes off your skin.
connect the tube to the rest of the apparatus.
e) Every 10 seconds, record the volume of oxygen until
the bubbles stop.
Results
If you sketch a quick graph, you may notice that the graph is not a
straight line. The rate of reaction is not constant; the reaction is
slowing down as the hydrogen peroxide is broken down.
The concentration of substrate seems to affect the rate of the

reaction.
Method B: Measuring the Rate with different concentrations of

substrate (Hydrogen Peroxide)
a) Measure five 0.1g samples of yeast powder using an electronic balance and keep them on
carefully folded pieces of paper (or in dry test tubes).
b) Measure 5 cm3 of each of five different concentration of hydrogen peroxide into five
separate boiling tubes. Label each tube.
c) Set up the inverted measuring cylinder as before.
d) Add the yeast to the first boiling tube and quickly connect the tube to the rest of the
apparatus.
e) After 1 minute record the volume of oxygen which has been produced.
f) Repeat for each of the five concentrations of substrate, hydrogen peroxide.
Results
If you sketch a quick graph, is the graph a straight line? Is the rate of
reaction constant?

2
Method C: Measuring the Rate with different pH of solution

a) Measure five 0.1g samples of yeast powder once more.
b) Measure 5 cm3 of 20Vol hydrogen peroxide into five separate boiling tubes.
c) Label each tube; pH2, pH4, pH6, pH8, pH10 and add five drops of buffer for each pH
d) Set up the inverted measuring cylinder as before.
e) Add the yeast to the first boiling tube and quickly connect the tube to the rest of the
apparatus.
f) After just 30 seconds, record the volume of oxygen.
g) Repeat for each of the five different pHs
Results
If you sketch a quick graph, does the shape show an “optimum” peak.
What is the pH of this?
Method D: Measuring the Rate at different temperatures

1. Measure five 0.1g samples of yeast powder once more.
2. Measure 5 cm3 of 20Vol hydrogen peroxide into five separate boiling tubes.
3. Label each tube; 10°C, 30°C, 50°C, 70°C and 90°C and put each boiling tube into a water bath
for 5 minutes. Check the temperature with a thermometer.
4. Set up the inverted measuring cylinder as before.
5. Add the yeast to the first boiling tube and quickly connect the tube to the rest of the
apparatus.
6. After just 10 seconds, record the volume of oxygen.
7. Repeat for each of the five temperatures of hydrogen peroxide.
Results
If you sketch a quick graph, what does the shape show?
Do you notice any change in the hydrogen peroxide solutions during

their treatment in the water bath?

3
Conclusion & Evaluation

Did your experiments show the expected patterns?
………………………………………………………………………………………………………………………………………………………….
………………………………………………………………………………………………………………………………………………………….
………………………………………………………………………………………………………………………………………………………….
………………………………………………………………………………………………………………………………………………………….
Describe any limitations to the accuracy of these experiments.
………………………………………………………………………………………………………………………………………………………….
………………………………………………………………………………………………………………………………………………………….
………………………………………………………………………………………………………………………………………………………….
………………………………………………………………………………………………………………………………………………………….
Suggest improvements for each limitation. The three diagrams below show different apparatus for
the same experiment. Why I each one an improvement on the experimental technique you have just
used?
………………………………………………………………………………………………………………………………………………………….
………………………………………………………………………………………………………………………………………………………….
………………………………………………………………………………………………………………………………………………………….
………………………………………………………………………………………………………………………………………………………….

4

Aims:: Experiment: Rates of Enzyme Controlled Reactions

Uploaded by

Copyright:

Available Formats

Aims:: Experiment: Rates of Enzyme Controlled Reactions

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Aims:: Experiment: Rates of Enzyme Controlled Reactions

Uploaded by

Copyright:

Available Formats

Experiment: Rates of Enzyme controlled reactions

A common way to do this in reactions

A simple inverted measuring cyllinder

 the concentration of substrate is higher

© David Faure, InThinking www.biology-inthinking.co.uk

Method A: Measuring the Rate of the Reaction

The concentration of substrate seems to affect the rate of the

Method B: Measuring the Rate with different concentrations of

© David Faure, InThinking www.biology-inthinking.co.uk

Method C: Measuring the Rate with different pH of solution

Method D: Measuring the Rate at different temperatures

Do you notice any change in the hydrogen peroxide solutions during

© David Faure, InThinking www.biology-inthinking.co.uk

Conclusion & Evaluation

Describe any limitations to the accuracy of these experiments.

© David Faure, InThinking www.biology-inthinking.co.uk

You might also like