Lab #11 – Fermentation

Lab #11 Fermentation

Introduction

All cells use glucose as their primary energy source. The hydrolysis of glucose releases energy
that can be used to synthesize ATP (adenosine triphosphate). The endergonic pathways
within the cell depend upon ATP to supply enough energy to keep them running.

Cellular respiration is the complete hydrolysis of glucose to carbon dioxide and water. This
reaction takes place in the mitochondria of eukaryotic cells.

C6H12O6 + 6O2 → 6CO2 + 6H2O + energy (36-38 molecules of ATP)

This reaction should look familiar-it is essentially the reverse of photosynthesis. But oxygen is
one of the reactants, so oxygen is necessary for this reaction to take place. This means it is an
aerobic (oxygen-requiring) reaction. Some organisms do not have oxygen-they live in low
oxygen environments like the deep sea or swamps. And some organisms are actually
poisoned by oxygen (obligate anaerobes). This tells us that there must be other pathways for
breaking down glucose.

Fermentation is an anaerobic reaction; it does not require oxygen. In a fermentation

pathway, glucose is only partially broken down. While there are many types of fermentation,
in this lab we will focus on ethanol fermentation. The yeast Saccharomyces cerevisiae is used
to make bread, beer, and wine. The process of ethanol fermentation goes like this:

C6H12O6 →2CO2 + 2C2H5OH + energy (2 molecules of ATP)

It is clear that fermentation is much less efficient than cellular respiration, and much less ATP is
produced by this process. However, it is the only method available to organisms that cannot
use oxygen.

(Left) S. cerevisiae, or common Baker’s yeast

(right) A fermenting bottle of strawberry wine. Notice the gases that are being produced (seen in the foam on top)

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In today’s lab, you will test the ability of yeast to ferment different sugars. In order to do this,
you will use a volumeter to determine how much carbon dioxide is being released by the yeast
cells. The different sugars we will test include lactose, sucrose, and glucose. Most of these
were previously covered in Lab #2 – Molecular Models. Glucose is a monosaccharide; sucrose
and lactose are disaccharides.

Pre-Lab Questions

1. Which is more efficient, cellular respiration or fermentation? Explain your answer.

2. Why would a cell use fermentation instead of cellular respiration?

3. What organism are we studying today?

4. What gas is produced by the fermentation process?
5. Which of the tubes is the control?
6. Which of the sugars that we are testing is/are disaccharides?
7. Which of the sugars that we are testing is/are monosaccharides?
8. Why are the tubes placed in a warm water bath?

9. What is the purpose of today’s lab?

Materials

• Small test tubes

• Larger test tubes
• Wax pencil
• Metric Ruler
• Suspension of live yeast cells

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• Pipettes
• Pipette Pump
• Distilled water
• 2% sucrose solution
• 2% glucose solution
• 2% lactose solution
• Warm water bath

Lab Procedures and Data

1. Us a wax pencil to mark the large outer tubes with the numbers 1, 2, 3 and 4. Also,
mark your test tubes with a group number or emblem to identify them in the warm
water bath.
2. Fill each SMALL test tube with 1 mL one of the following solutions:
a. Tube 1: glucose
b. Tube 2: lactose
c. Tube 3: sucrose
d. Tube 4: distilled water
3. The yeast suspension will be in the warm water bath. Use the pipette to thoroughly
mix the yeast before using it.
4. One at a time, fill each small test tube to the top with yeast. Slightly overfill
the test tube so that the solution is slightly bulging at the top.
5. Holding the small tube upright, invert the larger tube over it. Slide the small tube
into the large tube all the way until it touches the bottom of the large test tube (you
may need someone with long fingers or a pen or pencil to do this).
6. Quickly invert the tubes while holding them tightly together. This volumeter is
now ready for use.

The steps in setting up a volumeter to measure fermentation

From left to right: Step 4, step 5, and step 6

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7. The volumeter should have a very small air bubble at

the top. Use a wax pencil to mark the level of the air
bubble. Record the initial height of the gas bubble in Measure air
bubble
the table below. height
8. Repeat steps 1 to 7 until all of the volumeters are filled.
9. Place the volumeters in the test tube rack in the
warm water bath.
10. Wait for 20 minutes.
11. Record the final heights of the gas bubbles in the
table below.

Fermentation Experiment Results

Initial height of air Final height of gas Was fermentation
Tube Contents
bubble in mm bubble in mm occurring in this tube?
1 lactose 5mm 24 mm Yes

2 Glucose 4mm 5mm No

3 Sucrose 5mm 10mm Yes

4 Di water 3mm 4mm No

Which sugar was the yeast able to ferment most effectively? lactose
Was there any sugar(s) the yeast wasn’t able to ferment?
What was the purpose of the water?

What would it mean if the air bubble in the water volumeter increased in size?

Post-Lab Review Questions

1. What is the equation for ethanol fermentation?

C6H12O6 → 2 C2H5OH + 2 CO
2. Is fermentation aerobic or anaerobic? How could you tell?

3. Which gas was being produced inside the volumeters?

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 Lab #11 – Fermentation

4. Why didn’t the gas bubble in the water tube increase in size? _

5. How is cellular respiration different from ethanol fermentation? __

6. When making yeast bread, what gas causes the bread to rise?
Carbon dioxide

References
Brooker, Robert, Eric Widmaier, Linda Graham, and Peter Stiling. (2014).
Cellular Respiration and Fermentation. Biology 3rd Edition, 136-153.

Hoobler, Cynthia, Karen Duston, Adam Eiler, Jennie Plunkett, Kirsten Raines,
and Mary Wisgirda. (2007). Fermentation. General Biology I and II, 73-76.

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