Teacher Notes

Fermentation in Yeast
Objectives
Students explore two of the principle metabolic processes, aerobic and anaerobic cellular
respiration, used by facultative anaerobes to produce energy, and identify factors that affect
when alcohol fermentation (anaerobic cellular respiration) occurs in yeast. During this
exploration students investigate:
Byproducts produced from both aerobic cellular respiration and alcohol fermentation in yeast
Quantify the rate of both processes using sensors
Sugars fuel the fermentation process within yeast

Procedural Overview
Students gain experience conducting the following procedures:
Assembling a closed, sugar-rich, environment in which activated yeast cells can metabolize
Measuring oxygen gas and ethanol gas concentrations within the closed environment using
sensors
Associating each of the measurements with one of two metabolic processes occurring within
the yeast cells
Designing a short experiment that identifies whether lactose, maltose, or sucrose affects the
fermentation process within yeast

Time Requirement
Preparation time

25 minutes

Lab activity

90 minutes

Materials and Equipment

For each student or group:
Data collection system

Dry yeast solution, 1 L1

Oxygen Gas Sensor

0.5M Sucrose solution, 500mL2

Ethanol Sensor

1% Ethanol (for sensor calibration)

EcoChamber

Beaker, 500mL

Magnetic stirrer with stir Bar

3 other types of sugar (not sucrose)3

Beaker, 1,000mL

To formulate activated dry yeast using dry yeast and water, refer to the Lab Preparation section.

To formulate 0.5M Sucrose solution using Sucrose and water, refer to the Lab Preparation section.

Refer to the Lab Preparation section for alternative sugar suggestions.

012-12307A

Fermentation in Yeast
Safety
Add these important safety precautions to your normal laboratory procedures:

Ensure that you understand and adhere to safe laboratory practices when performing
any activity in the classroom or lab.

Use personal protective equipment such as safety glasses or goggles, gloves, and aprons
when appropriate.

Background
Are yeast aerobic or anaerobic organisms? Yeast are actually single-celled organisms that are
facultative anaerobes, organisms that have the ability to undergo aerobic respiration in the
presence of oxygen and anaerobic respiration in its absence. With oxygen present, yeast will
preferentially undergo aerobic respiration because they can make 36 ATP molecules per glucose
molecule through aerobic respiration, compared to the 2 ATP molecules produced through
anaerobic respiration.
In this activity, yeast will use the dissolved oxygen and the sugar in the water solution as the
reactants in the equation:
C6H12O6 + 6O2 6 CO2 + 6H2O + energy (primarily stored in ATP molecules)
As time progresses, you should see a decrease in concentration of oxygen because the yeast cells
use it to create ATP. As oxygen becomes less available the yeast will change their metabolic
pathway to alcohol fermentation to continue producing the ATP required for cellular processes.
C6H12O6 2CO2 + 2C2H6O + energy (primarily stored in ATP molecules)
The fermentation process does not produce ATP directly but allows NAD+ to be recycled into
NADH which allows glycolysis to continue. The net yield for alcoholic fermentation is only 2
molecules of ATP per glucose molecule. As more cells begin to ferment, the concentration of
ethanol in the chamber will increase. This fermentation process is similar in many ways to lactic
acid fermentation which occurs in muscle cells when oxygen levels are low. Most students will
have direct experience with lactic acid fermentation because it is largely responsible for the
burn felt in muscles following intense exercise.

Lab Preparation
Before the Lab
1. Prepare the yeast solution at least 30 minutes before the lab begins. To make 1 liter (L) of
solution, add 20 grams (g) of dry bakers yeast to 1 L of warm water (30 to 35 C). Alter the
proportions if you need to make more than 1 L. Bakers yeast from grocery stores works very
well, but be sure to check the expiration date. Yeast purchased in bottles of 100 g or larger
are widely available and are often more economical than packets. Once you have made the
initial solution, it can be stored at room temperature until ready for use. Just make sure that
it stays between 20 C and 45 C.
2. To prepare the 0.5 M sucrose solution:
a. Measure 171 g of sucrose (table sugar).
b. Place the sugar in a beaker or bottle.
c. Add distilled water to make 1 liter of solution.

Teacher Notes
One liter is enough for two groups, so alter the proportions so that each group receives
500 mL of solution. You can store the solution at room temperature.
3.

You will need 1% ethanol solution to calibrate the ethanol sensor.

4.

To prepare the ethanol sensor for use, connect it to a data collection system that has been
plugged into an electrical outlet before starting the lab. The sensor will warm up and
perform optimally during the lab.

5.

Have at least three other types of sugar available to students for the inquiry part of the lab.
Lactose, maltose, and glucose are excellent choices as they will provide a variety of results.
Be sure that you have enough for each group to make at least a 0.5 M 500 mL solution of the
sugar they choose.

Sample Data
The following screenshots display examples of data similar to what students should expect to
see.

Fermentation in Yeast

Answer Key
Analysis Questions
1. Did the O2 concentration in the chamber increase or decrease? Does this indicate that
respiration occurred? Explain why or why not?
The O2 concentration in the chamber decreased 4% (student numbers may vary slightly, but they should cite
collected data in their response) during data collection period. This indicates that oxygen was being consumed
by the yeast cells as they conducted aerobic respiration. If the yeast cells had been creating ATP solely through
anaerobic respiration, no change in oxygen levels would be observed.

Teacher Notes
2. If you were to allow the yeast sample to sit for a very long time, do you think the O2
concentration would eventually go to zero? Explain why or why not.
While theoretically the oxygen concentration would drop to zero if the yeast respired over a long enough period
of time, the data suggests that below a certain concentration the yeast are primarily conducting alcohol
fermentation and oxygen consumption stabilizes.

3. Did the amount of ethanol in the chamber increase or decrease? Does this indicate that
fermentation occurred? Explain why or why not?
The amount of ethanol in the chamber increased 0.8% (student numbers may vary slightly, but they should cite
collected data in their response) during data collection period. The presence and gradual increase in the amount
of ethanol in the chamber indicates that the yeast are conducting alcohol fermentation. The rate of ethanol
production should increase over time as oxygen becomes less available for aerobic respiration.

4. Based on our data, what conclusion(s) can we draw in terms of whether yeast are facultative
anaerobes? Support your conclusion using your data.
In the lab we observed evidence of yeast cells conducting aerobic and anaerobic respiration, suggesting that
they are facultative anaerobes. We observed a decrease in the oxygen concentration, which suggests the yeast
cells were aerobically respiring. As the amount of available oxygen decreased there was an increase in the
ethanol concentration, which suggests the yeast was conducting alcohol fermentation.

Design Your Own Experiment (Sample Responses)

Students are encouraged to develop their own inquiry based on questions that have arisen in the
course of completing the guided portion of this lab. The following sections are sample responses
that students may develop as well as suggestions and activity ideas.
Question
Students should write their question so it includes the independent and dependent variable(s)
they will monitor. Some investigable questions include:
If the sugar was changed to (lactose, maltose, etc.), would alcohol fermentation still occur?
Would one type of sugar make fermentation happen more rapidly, or create more ethanol, than
the other types?
Prediction
After conducting the guided portion of this lab, students have enough information to formulate a
hypothesis that develops a cause and effect relationship between an independent variable and
dependent variables, which should answer the questions: What do you think will happen? Why
do you think so? One example of a hypothesis is shown here:
We predict that if the sugar is changed to lactose, alcohol fermentation will occur because, like
sucrose, lactose is a disaccharide.
Materials
Many inquiries can be completed with the original materials for the guided portion of this lab.
Additional materials may include:
Lactose, maltose, glucose, fructose, and other sugars
Procedure
Students may add pages of text boxes and write their procedure in the data collection system,
they can write their procedure in a lab notebook, or they can process their procedure in a
computer file and submit it to you electronically. Whatever way the procedure is prepared, it

Fermentation in Yeast
should conform to certain guidelines and should be approved by you before the students begin
their inquiry. Also, clarify the safety parameters for their investigations before they begin.
Guidelines include the following:
The procedure should be a numbered list.
Sentences should be concise, well-written, complete, and effectively communicate each step.
Both independent and dependent variables should be described, and students should indicate
how the independent variable will be changed and how the dependent variables will be
measured.
Additional controlled variables that are to remain constant should be described.
The method of displaying collected data is identified. Students choose a graph, table, digits
display or meter.
Students should record observations while they collect data.
Building a Page for Data Collection
Instructions are provided for building a page that includes a table, graph, digits display, or
meter.
Students begin by selecting Add New Page
. From there, they can build a page
for data collection or for writing notes and observations. To collect data they must
select a sensor and then select the type of display they want to use: table, graph,
digits display or meter.
To record observations or write notes, students choose a large text box
. They will be able to
enter text on the entire page. Another option is to place a data collection box, such as a graph
and two small text boxes, on the same page. Students should experiment with different design
elements until they find an arrangement that suits their purpose.
Analyze Your Data
Students will analyze their data, including any mathematical and graphical analysis, such as
finding the mean or the percent increase.
Draw Your Conclusion
Students should state whether or not their hypothesis was correct. They should summarize how
a change to their independent variable caused a change of their dependent variable(s) in a
predictable way.
Make the Connection
Students take what they have learned in their own experiment and add it to what they have
learned from the guided portion of the lab to answer the overarching challenge of the lab.
When the sugar was changed did the yeast cells continue to exhibit an ability to perform
anaerobic and aerobic respiration?
How did the results of your experiment compare to the guided experiment?

Teacher Notes

Your Experiment Assessment Rubric

Lab Sections

A Good Answ er

A Better Answ er

The Best Answ er

Written as an exploration or
testable question

Written as a question;
includes at least one variable

Written as an ifthen question;

Reflects what students

expect to happen

Reflects what students

expect to happen and why

Merges the driving question

Procedure

Has teacher approval

Has teacher approval

Has teacher approval

Materials list

Students use the same

At least one essential new

Refers to materials used in a

materials they used in the

first part of the lab.

material listed

different way or lists new

materials

Numbered list

Numbered list, well-written

Numbered list, concise, clear

Driving Question

Hypothesis

Clear directions

includes both the independent

and dependent variables

with what students expect to

happen and why

steps; thorough

Independent,
dependent, and
controlled
variables

Identifies the independent,

Identifies and explains how

Identifies and explains how

dependent, and controlled

variables

variables will be controlled or

measured

variables will be controlled or

Sample rate and

sensor

Includes sample rate or

Includes sample rate and

Includes sample rate, sensor

sensor

sensor setup

setup, and how it will be used

Observations

Includes an observation for

Observes both independent

Describes behavior of both

each run

and dependent variables

during each run

independent and dependent

Collected one run of data

Collected multiple runs of

Collected multiple runs of data;

shown on a graph or in a

data; axes or columns are

axes or columns are correctly

table; axes or columns are

correctly labeled

correctly labeled; conditions

are correctly labeled

labeled, conditions correctly

Some mathematical or

Pertinent calculations, such

Pertinent calculations, such as

graphical analysis is shown

or with a statistics tool

as the mean or change in

the mean or change in value,

value, are calculated and

visually displayed

are calculated and different

Explains the relationship

Uses data to explore the

Uses graphs and tables to

between the independent

and dependent variables

cause and effect relationship

explore the cause and effect

between the independent

and dependent variables

relationship between the

Answers the conclusion

Answers the conclusion

More extensively answers the

question and states whether

question and states whether

conclusion question and states

or not their hypothesis was

correct

or not their hypothesis was

whether or not their hypothesis

correct and answers their

(driving) question

was correct and answers their

(driving) question

Analyze Data

measured, and how the

independent variable will be
changed

variables and all conditions

during the run

labeled; pertinent values or

trends indicated

Analysis
Mathematical or
graphical
analysis

Conceptual
analysis

Conclusion

conditions are compared

visually

independent and dependent

variables