Sugar Fermentation in Yeast

Yeast can metabolize some foods, but not others. For an organism to make use of a potential
source of food, it must be capable of transporting the food into its cells. It must also have the
proper enzymes capable of breaking the food’s chemical bonds in a useful way. Sugars are vital
to all living organisms. Yeast are capable of using some, but not all sugars as a food source.
Yeast can metabolize sugar in two ways, aerobically, with the aid of oxygen, or anaerobically,
without oxygen.
In this lab, you will try to determine whether yeast are capable of metabolizing a variety of
sugars. Although the aerobic fermentation of sugars is much more efficient, in this experiment
we will have yeast ferment the sugars anaerobically. When the yeast respire aerobically, oxygen
gas is consumed at the same rate that CO2 is produced—there would be no change in the gas
pressure in the test tube. When yeast ferment the sugars anaerobically, however, CO2 production
will cause a change in the pressure of a closed test tube, since no oxygen is being consumed. We
can use this pressure change to monitor the fermentation rate and metabolic activity of the
organism.
The fermentation of glucose can be described by the following equation:

C6H12O6 → 2 CH3CH2OH + 2 CO2 + energy

glucose ethanol carbon dioxide

Note that alcohol is a byproduct of this fermentation.

OBJECTIVES
In this experiment, you will
 Use a Gas Pressure Sensor to measure the pressure change caused by carbon dioxide
released during fermentation.
 Determine the rate of fermentation.
 Determine which sugars yeast can metabolize.

Figure 1

MATERIALS
computer 18 × 150 mm test tube
Figure 1
 Vernier computer interface 1 L beaker (for water bath)
Logger Pro basting bulb or Beral pipette
Vernier Gas Pressure Sensor hot and cold water
l-hole rubber stopper assembly test tube rack
plastic tubing with Luer-lock fitting thermometer
5% glucose, sucrose, lactose, and one yeast suspension
other sugar solution vegetable oil in dropper bottle
ring stand utility clamp

PROCEDURE
1. Connect the plastic tubing to the valve on the Gas Pressure Sensor.

1. Connect the Gas Pressure Sensor to the data-collection interface, and then connect the
interface to your Chromebook, computer, or mobile device. Launch Graphical Analysis.

2. Prepare a water bath for the experiment. A water bath is simply a large beaker of water at a
certain temperature. This ensures that the yeast will remain at a constant and controlled
temperature.

a. To prepare the water bath, obtain some warm and cool water from your teacher.
b. Combine the warm and cool water in the 1 L beaker until it reaches 38–40°C. The beaker
should be filled with about 600–700 mL of water.
c. Place the thermometer in the water bath to monitor the temperature during the
experiment.
4. Obtain two test tubes and label them 1 and 2.

Figure 2
 5. Your team will test two of the four sugar solutions. Obtain two of the four sugar solutions:
glucose, sucrose, lactose, and fructose solution, as directed by your instructor.

6. Place 3.0 mL of the first sugar solution into test tube 1 and 3.0 mL of the second sugar
solution into test tube 2. Record which solutions you tested in Table 1.

7. Pipet 3.0 mL of yeast suspension into test tube 1. Gently swirl the test tube to thoroughly mix
the yeast into the solution. Important: The yeast suspension must be removed from the
middle of a yeast source that is being stirred by a magnetic stirrer at a constant stirring speed.

8. In test tube 1, place 1 mL of vegetable oil to completely cover the surface of the
yeast/glucose mixture as shown in Figure 2. Note: Be careful to not get oil on the inside wall
of the test tube.

9. Insert the single-holed rubber-stopper into the test tube. Note: Firmly twist the stopper for an
airtight fit.

10. Connect the free-end of the plastic tubing to the connector in the rubber stopper as shown in
Figure 3.

Figure 3
11. Set test tube 1 in the water bath and secure it with a utility clamp and ring-stand as shown in
Figure 1. Note: Be sure that most of the test tube is completely covered by the water in the
water bath. The temperature of the air in the test tube must be constant for this experiment to
work well. Be sure to keep the temperature of the water bath constant.

12. Click or tap Collect to start data collection. Note: Maintain the temperature of the water bath
during the course of the experiment. If you need to add more hot or cold water, first remove
about as much water as you will be adding, or the beaker may overflow.

13. Data collection will end after 15 minutes. Monitor the pressure readings displayed on the
screen. If the pressure exceeds 130 kilopascals, the pressure inside the tube will be too great
and the rubber stopper is likely to pop off. Disconnect the plastic tubing from the Gas
Pressure Sensor if the pressure exceeds 130 kilopascals.

14. When data collection has finished, a graph of pressure vs. time will be displayed.

15. Disconnect the plastic tubing connector from the rubber stopper. Remove the rubber stopper
from the test tube and discard the contents in a waste beaker.

16. Determine the rate of fermentation:

a. Select the data in the sloping portion of the graph.

 b. Click or tap Graph Tools, , and choose Apply Curve Fit.
c. Select Linear as the curve fit. Click or tap Apply.
d. Record the slope of the line, m, as the rate of fermentation in Table 1.
e. Dismiss the Linear Curve fit box.

17. Repeat Steps 7–16 using test tube 2. Note: The previous data set is automatically saved.

DATA

Table 1

Type of Sugar Rate of Fermentation (kPa/min)

Test tube 1

Test tube 2

Table 2: Class Averages

Sugar Tested Fermentation Rate

(kPa/min)

Control

PROCESSING THE DATA

1. Share your data with the rest of the class by recording the sugar type you tested and the rate
of fermentation on the board.

2. Using the class data, calculate the average rates of fermentation for each of the sugar types
tested. Record the average rates in Table 2, along with the names of the 4 sugars tested.

3. On Page 2 of the experiment file, use the class data in Table 2, make a bar graph of rate of
fermentation vs. sugar type. The rate values should be plotted on the y-axis, and the sugar
type on the x-axis.
Claim:
Write a statement that
responds to the question.
● Does the claim
answer the
question?
● The claim should
only answer the
question. Does
the claim include
an explanation or
reasoning?
● Is it more than a
“Yes” or “No”
answer?
● Is the claim
written in
complete
sentences?

Evidence:
Provide scientific data to
support your claim. Your
evidence should be
appropriate (relevant)
and sufficient (enough to
convince someone that
your claim is correct).
Feel free to use bullet
points instead of
sentences.
● Is each piece of
evidence relevant
to the claim?
● Is a property
labeled data table
and graph
included?
● Is there
qualitative data
included?
● Is there enough
evidence listed to
support the
claim?
● The evidence
should lack an
 explanation or
reasoning. Is
each piece of
evidence strictly
an observation or
data from the lab?
● Is the evidence
listed specific and
not vague or a
generalization?

Reasoning
Use scientific principles
and knowledge that you
have about why your
evidence (data) supports
your claim. In other
words, explain how your
data proves your point.
(Paragraph format)
Need help? Click here.

● Is there a
justification of
how each piece of
evidence
supports the
claim?
● The reasoning
should not simply
repeat the
evidence. Is
there a link
between the
evidence and
claim provided?
● Is the science
principle
explained why
the evidence
supports the
claim?
● Is the reasoning
written in
complete
sentences?
QUESTIONS
1. Considering the results of this experiment, can yeast utilize all of the sugars equally well?
Explain.

2. Hypothesize why some sugars were not metabolized while other sugars were.

3. Why do you need to incubate the yeast before you start monitoring air pressure?

4. Yeast live in many different environments. Make a list of some locations where yeast might
naturally grow. Estimate the food sources of each of these locations.