Investigation Into the Effect of Temperature On the Rate of Respiration of Yeast

Investigation Into the Effect of Temperature On the Rate of Respiration of Yeast

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Investigation Into the Effect of Temperature On the Rate of Respiration of Yeast

Preliminary Work

For my preliminary work, I am working with 35ml of yeast. I think that
this is the best volume to use as it is about ¾ of a test tube full,
and it allows for the yeasts expansion when heated. I am trying to
find out the best range of temperatures to be used in finding out the
respiration of the yeast, and I am also trying to find an
equilibration time that can be used in the main experiment, as the
time taken for the yeast to heat up to the desired temperature.

Apparatus

- One beaker

- Two test tubes

- Delivery tube with bung

- Yeast (35ml)

- Water

- Stopwatch

- Thermometer

- 35ml syringe

Method

1. A beaker was filled with water then heated to the desired
temperature.

2. A test tube was then filled with 35ml of yeast and placed in the
beaker of water.

3. The time taken for the yeast to heat up to the temperature of the
water in the beaker was then measured using a stopwatch and
thermometer.

4. A delivery tube with a bung was then placed into the test tube of
yeast, with the bung end of the tube going into the test tube of
yeast.

5. A second test tube was then filled with water and placed in a test
tube rack.

6. The other end of the delivery tube was then placed into this second
test tube.

7. The number of bubbles coming out of the delivery tube into the
second tube per minute was then measured.

Results – Equilibration time to be used = 4 minutes

Temperature (oC)

No. of Bubbles per min

Equilibration Times

30

55

2 mins 3 secs

40

43

2 mins 36 secs

50

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Investigation Into the Effect of Temperature On the Rate of Respiration of Yeast

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29

3 mins 12 secs

60

24

3 mins 43 secs

70

18

4 mins 1 second

3


Main Experiment

Introduction

In this experiment I am trying to find out what the effect of raising
or lowering the temperature has on the respiration of yeast. I will
use my preliminary work as a guide to my final experiment, by taking
the results from my preliminary work and using the things that I have
found best to use, in this experiment. I am using the equilibration
time of 4 minutes because it is the longest time out of all of them
and it wouldn’t work for the 70o temperature if it was only 2 minutes.

However, I will not be using the bubbles method in my main experiment
as it is far too inaccurate and does not provide the right results.
Instead I will be using a method of displacement. As in the
preliminary work, there will be a second test tube of water, but
instead of counting the bubbles in this water, I will be measuring the
amount of water displaced by the gas (carbon dioxide) coming from the
yeast.

I expect to see two types of respiration from the yeast; aerobic (with
oxygen) and anaerobic (without oxygen). Here are the two equations
representing this:

Aerobic:

Glucose + Oxygen Energy + Carbon Dioxide + Water

C6H12O6 + 6O2 Energy + 6O2 + H2O

Anaerobic:

Glucose A bit of energy + Ethanol + Carbon Dioxide

C6H12O6 A bit of energy + 2C2H5OH + 2CO2

I am going to use the method of displacement for my main experiment
because it provides much more accurate results, and as shown in the
equations, yeast produces the gas carbon dioxide when it respires. I
expect to see both types of respiration because, the yeast at the top
of the test tube will have access to oxygen, and can respire
aerobically, while underneath, the yeast has no access to oxygen and
will have to respire anaerobically. The carbon dioxide produced from
the yeast’s respiration will displace some of the water in the test
tube, and I will be able to measure the amount of water displaced to
get the results for my experiment.

Prediction

From the results of my preliminary work, I predict that between 20o
and 40oC, the yeast enzymes will be respiring fastest because enzymes
work best at room temperature. I also predict that as the temperature
goes up, the respiration will get slower and slower because the
enzymes will start to denature (the yeast enzymes stop working) at
higher temperatures. This is shown in the graph on the next page:

4


40o is the optimum temperature because it is just the right
temperature for the enzymes to be working at maximum rate. Yeast is an
enzyme, which means that it is also a catalyst. Enzymes work using the
‘lock and key’ theory, where the ‘key’ fits into the active part of
the enzyme (the ‘lock’) and the reaction takes place. The key then
unlocks to form one or two more new substances and the enzyme is ready
to bind with another of these substances. An enzyme can only bind with
a substance that fits the shape of the active part of the enzyme, so,
because the enzymes are sensitive to higher temperatures, the active
site on the enzyme changes shape so much that binding can hardly take
place. This is the denaturation. Up until a temperature of about 50o,
as the temperature rises, the yeast works at a faster rate with the
heat, because catalyst speed up reaction rate, but it is heat that
speeds up the catalyst. This is shown in the diagram below:

5


Apparatus

- A test tube

- Two beakers

- Water

- Yeast

- Stopwatch

- Delivery tube

- Thermometer

- Measuring cylinder

- 35ml syringe

Method

1. Heat up the water in the water bath to the desired temperature,
using a tap water to cool, and a thermometer to measure the
temperature.

2. Fill a beaker with water of the desired temperature (30, 40, 50,
60, 70oC) measuring the temperature using a thermometer.

3. Fill the test tube with 35ml of yeast using the syringe and place
into the beaker of water.

4. Measure the equilibration time (4 minutes) using a stopwatch and
check using a thermometer.

5. During the 4 minutes, fill a measuring cylinder to the top mark.

6. When the equilibration time is up, place the bung end of the
delivery tube into the test tube of yeast, and the other end of the
delivery tube into the measuring cylinder. Turn the measuring cylinder
upside-down taking care not to lose water, and place into the other
beaker, filled with cold water.

7. Measure 2 minutes using the stopwatch, and take out the bung from
the test tube of yeast.

8. Measure the displacement of the water in the measuring cylinder.

9. For each of the temperatures, repeat the experiment twice.

See experiment diagram next page.

2


Preliminary Diagram

6



Main Experiment Diagram

7


Results

Temperature (oC)

Displacement

(ml)

Repeat

Temp. (oC)

Displacement

(ml)

Average

Displacement

(ml)

30

2.0

30

2.5

2.25

40

7.5

40

6.5

7.0

50

4.5

50

4.0

4.25

60

3.0

60

3.0

3.0

70

1.0

70

1.5

1.25

Equilibration time used= 4 minutes

See graph on next page

Conclusion

From the graph we can see that at 30oC, the displacement (respiration
indicator) is 2.25ml, and this is quite a low level of respiration.
The respiration reaches its optimum temperature at 40oC and the level
of respiration falls as the temperature rises and the enzymes
denature. At 70oC however, the highest temperature used, there is
still some respiration going on, and this was not expected, as it was
thought that the enzymes would have completely denatured (stopped
working).

At temperatures around 40oC, the optimum temperature, there are more
collisions between the enzymes and glucose molecules, and the
molecules are slowly broken down and used for respiration. When the
temperature is too high, such as 70o, the enzymes start to denature,
which means that there are less and less collisions with the glucose
molecules, thus less respiration.

Enzymes work using the ‘lock and key’ theory, where the ‘key’ fits
into the active part of the substrate, and a reaction takes place. The
key then unlocks to form one or two other substrates, and the enzyme
is ready to bind again with one of these new substrates. An enzyme can
only bind with a substrate that fits the shape of the active part of
it, so, because the enzymes are sensitive to higher temperatures (in
my graph any temperature below around 50oC) the active part of the
enzyme changes shape so much that the binding can hardly take place.
This is the denaturation, and it also means less respiration. As the
temperature rises to 40oC, the yeast enzyme works at a faster and
faster rate, because it is a catalyst and therefore speeds up reaction
rate. This ‘lock and key’ theory is shown in the diagram below:

9

11B

Analysis

My conclusion does support my prediction to quite a large extent. The
graphs both show that I predicted correctly the optimum temperature
would be 40oC, and also that after this temperature, the enzymes would
start to denature. At 40oC, the enzymes are working the fastest,
colliding with the glucose molecules, and breaking them down to be
used for respiration. The only difference between the prediction and
conclusion is that I predicted that the enzymes would have completely
denatured by 70oC, when in fact there was still some respiration. This
shows that my conclusion supports my prediction to quite a large
extent.
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