INVESTIGATION 4.

5
Anaerobic fermentation in yeast
STUDENT LABORATORY NOTES
Abubaker Nassief

Introduction
Yeast, like all living organisms, must produce ATP to provide the energy for cellular functions. When
oxygen is available, yeast undergo aerobic respiration. When no oxygen is present, yeast undergo
anaerobic fermentation.
In both aerobic respiration and anaerobic fermentation, glucose is required as the initial input. Like plants,
when yeast undergoes anaerobic fermentation, ethanol and carbon are produced. Limewater can be used to
measure carbon dioxide. When limewater and carbon dioxide react, a milky precipitate is produced,
making the solution appear cloudy.

Aim
To observe the rate of fermentation in yeast under different glucose concentrations

Hypothesis

It is hypothesised that as the amount of glucose used increases, the respiration growth will
increase in correlation, it also hypothesised that as fermentation occurs, the

CO2 being produced goes through the connecting tube and produces bubbles

Preparation for task

Location to conduct this task
In the laboratory
Duration
50 minutes
Group size
2 students per group (may be up to 4 students depending on class size and resources)

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 Topic 4 | Photosynthesis, cellular respiration and biotechnological applications

Methodology
Materials (per group)
 Yeast (either 15 mg of Baker’s yeast or 2 packages of dried yeast).
 Warm water
 Glucose (0 g, 5 g, 10 g, 20 g and 30 g)
 Electronic balance
 Test tube or conical flask with stopper and glass tube (a conical flask is recommended as this
is the vessel in which anaerobic fermentation will occur).
 Test tube (a medium to large sized test tube is recommended)
 Test tube rack
 Limewater
 Beaker
 Measuring cylinder
 Thermometer
 Laminated sheet with grades of black line
 Stopwatch
 Kettle (to share)

Health and safety guidelines

 Steam burns can occur from kettles. Keep arms and eyes away from the spout. Run hands
under cold water if burnt, eyes 15 minutes under the eye wash if affected.
 Any food materials should not be ingested in a laboratory.
 Glass tubing can be fragile, handle with care and do not force tubes into stoppers as they may
snap.

Method
1. Add limewater to a test tube. There should be enough limewater in the test tube to match the
height of the laminated card (leaving some space at the top to prevent overflow).
2. Heat water with a kettle, pour approximately 150 mL into a beaker with a thermometer; and
allow to cool to 50 oC.
3. Using a measuring cylinder, carefully measure 100 mL of 50 oC water and place into a
conical flask (Note: a test tube, as shown in the figure on the next page may instead be used.
If a test tube is used, the quantities and volumes should be reduced.).
4. Sprinkle 1 sachet (or 2 tsp) of yeast on top. Measure the appropriate amount of sugar (your
teacher may designate this), add to the yeast solution and stir gently. Attach the stopper with
the glass tube, connecting the glass tube to the test tube containing limewater, as shown in the
figure (Note: while the figure shows a test tube as the vessel for the yeast and glucose, this
may be a conical flask, as outlined in step 4. You may also have more limewater, depending
on the size of the test tube.)

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 Topic 4 | Photosynthesis, cellular respiration and biotechnological applications

5. Start a timer. Hold the laminated colour card behind the test tube with the limewater each
minute for 10 minutes. Record the highest number you can see through the solution in the
results table.

6. Collate class data for the investigation.

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 Topic 4 | Photosynthesis, cellular respiration and biotechnological applications

Results
Complete the following results table showing the data obtained in the investigation.
TABLE 1 The highest number visible through the limewater solution each minute

Number of lines observed in different amount of sugar (g)

Time (mins) 0g 5g 10 g 20 g 30 g

1 5 5

2 5 5

3 5 5

4 5 5

5 5 4

6 4 4

7 3 3

8 3 2

9 2 1

10 1 1

Discussion questions
1. The results were obtained by observing how transparent the solution was using a chart with a
gradient of colour.
a. Explain, with reference to the results, what type of data was collected?

In the results recorded above, the data used is quantitative data because a chart with
quantities was used to represent the results of the experiment, and observe
color/transparency of limewater and the results that were made
under observation. 5 being easiest to see through and 1 being hardest to
see through

b. Identify a limitation in this method of data collection and suggest a way that more
accurate results could be obtained.
Limitations include:
-Temperature doesn’t remain constant as in the beginning of the procedure it was above
50 degrees and we were waiting a along time for it to settle at 50 degrees, furthermore it start
decreasing overtime which could effect the rate of CO2 released.
 - Laminated sheet with grades of black line wasn’t available in the Materials provided when it
might’ve possibly had a significant part of the experiment.
 colour/shade varies for everyone therefore it could be Unreliable due to the method of
observation used therefore it cannot be 100% accurate.

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 Topic 4 | Photosynthesis, cellular respiration and biotechnological applications

Improvements:
 - Change method of observation to something more accurate
 - Keep temperature constant with the help of a water bath or a Bunsen burner


2. What does the colour of the limewater infer about the rate of anaerobic fermentation in yeast?

Lime water is used in fermentation, to effectively, visually determine whether

carbon dioxide was produced and how fast. When lime water reacts with CO2
it becomes a cloudy milky liquid. However, through the observing of how fast
the limewater turns cloudy the rate on anaerobic fermentation can be shown.

3. This practical investigated anaerobic fermentation in yeast. Outline the difference you would
expect in the products if anaerobic fermentation in animal cells was investigated.
Pyruvate broken down after glycolysis would result in lactate or lactic acid if animal cells
were to be tested.. No carbon dioxide would be produced.
They differ due to differences in the enzymes and reaction pathways between
the two types. Animals have an enzyme called lactate dehydrogenase that
directly transforms pyruvate into lactic acid. That enzyme is absent in plants or yeast in this case.

4. This practical investigated the effect of glucose on the rate of anaerobic fermentation. Predict
what effect temperature would have on the rate of anaerobic fermentation.
Temperature is a very important factor in yeast fermentation. Since yeast is a
living organism is must have suitable and optimal conditions to thrive. The
temperature directly affects the type of fermentation prodcued. As the
temperature gets higher, the yeast will produce more carbon dioxide, until at
some point carbon dioxide production will decrease, that is when the yeast cells
have become denatured due to the increase in temperature beyond its optimum
range.
Conclusion
Write a conclusion for this investigation.
The aim of this experiment was to observe the rate of fermentation in yeast under different glucose
concentrations. After conducting this experiment we can say that the hypothesis was successful in
supporting the aim as the time taken for the limewater to go cloudy was observed; from this it was
concluded that the faster the time taken to turn the limewater cloudy, the
faster the rate of respiration. The hypotheses also states that as fermentation occurs, the
CO2 being produced goes through the connecting tube and produces bubbles out
through the lime water was also proven right . Overall, the hypothesis for this
experiment stating, ‘As the amount of glucose is increased, the respiration
rate will increase’ and “ as fermentation occurs, theCO2 being produced goes through the connecting
tube and produces bubbles was supported as can be seen by the data produced in the

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 Topic 4 | Photosynthesis, cellular respiration and biotechnological applications

table above and from the visual results provided.

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