Activity 1.

Cellular Respiration (Carbon Dioxide Production)

Our cells need a continuous supply of energy to sustain a myriad of diverse functions that sustain
life. Our cells get this energy primarily by breaking down a fuel source (e.g. glucose) to carbon
dioxide through a multistep process referred to as cellular respiration. Cellular respiration has
three broad steps; glycolysis, the Krebs cycle and the electron transport chain.

Glycolysis: Glycolysis is also known as the “EMP” pathway after the three Nobel Laureates who
elucidated it - Embden, Meyerhof and Parnas. Glycolysis is a sequential process of breaking
bonds and altering glucose in a sort of disassembly line pathway. The 6-carbon glucose molecule
(C6H12O6) is reconfigured by a set of enzymes into a 6-carbon sugar called fructose, which then
picks up a pair of phosphate groups making it fructose-di-phosphate (FDP). Then the FDP is split
in half to form two trioses (3-carbon molecules), which eventually form two 3-carbon pyruvates
(C3H6O3). The phosphates are released to form ATP and the pyruvates can then be further broken
down. There are three major pathways that the pyruvates can follow depending upon the oxygen
levels in the cell and the enzymes present.

In an aerobic environment, the presence of oxygen (O2) and enzymes allows glucose to be broken
down all the way to CO2. The two pyruvates from glycolysis can be broken down to form two CO 2
molecules and two 2-carbon Acetyl co-A molecules which can then enter the mitochondria. In
the mitochondria, Krebs cycle and the electron transport chain proceed catalyzed by many
different enzymes. Ultimately, 34 to 36 ATP molecules form and free CO 2 is released. So, we
inhale O2 to be used in electron transport chain, and we exhale CO2 as a waste product of cellular
respiration.

Aerobic cellular respiration

C6H12O6 + 6O2 → 6H2O + 6CO2 + 36 ATP

Glucose + Oxygen Water + Carbon dioxide + Energy

In a microaerobic environment, oxygen depletion in our muscle cells causes glucose to be

converted into lactic acid. Lactic acid is released from the oxygen depleted cells to the blood
stream to be slowly neutralized by liver enzymes in the 24 hour period after strenuous exercise.

In an anaerobic environment, due to the total absence of oxygen, certain species of fungi and
bacteria with special enzymes allow them to ferment pyruvates into a variety of products. For
example, fungal yeasts have enzymes that form ethanol and CO 2 from pyruvate. Ethanol is the
alcohol in beer and wine. The carbon dioxide produced by yeasts in bread can make the bread
rise or produce the carbonation of beer. Propioniobacter produce the large gaseous CO2 bubbles
seen in Swiss cheese. Acetobacter have special enzymes that can catalyze acetic acid formation,
which can sour wine and change it into vinegar. Lactobacillus can produce lactic acid which can
sour milk and be used to make yogurt and cheese curds. Methanogens can produce methane gas
(CH4). These processes are referred to as anaerobic fermentation

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 Anaerobic fermentation in yeast and bacterial cells

C6H12O6 → 2C3H6O3 → CO2 and/or Other products

Glucose Pyruvate Carbon dioxide

Cellular Respiration Activity

http://www.bch.cuhk.edu.hk/vlab2/animation/fermentation/index.html

Objectives:
● To observe respiration in yeast and the release of carbon dioxide
● To observe the effect of heat on anaerobic fermentation
● To observe a change in pH in carbonated water

Virtual Materials:
● Yeast solution
● Glucose
● Flour
● 5 flasks
● 5 balloons
● Heater
● Water
● Red and blue litmus paper (about: https://sciencing.com/difference-blue-red-litmus-
paper-8731865.html)

Procedure – use the online virtual lab “Yeast Fermentation” to outline the experimental steps
(5 points)
1. Prepare flask one with water and yeast
2. Prepare flask 2 with yeast, water, and glucose
3. Prepare flask three with yeast, glucose, and water. Boil it in the heater for 3 minutes
4. Prepare flask 4 with glucose and water
5. Prepare flask five with water, yeast, and flour
6.

Hypothesis and Prediction.

Under which experimental condition(s) you’d expect the balloon to inflate? Write a hypothesis
for this experiment. What are your independent and dependent variables? (5 points)

If you put a balloon over the yeast, glucose, water, and boiling flask the balloon will inflate.
The independent variable is the yeast, glucose, flour. The dependent variable is the balloon

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Record your observations by filling in Table 1 and Table 2.

Table 1: Gas formation in the flasks (10) points).

Flask #1 Flask #2 Flask #3 Flask #4 Flask #5

Yeast (+/-) Yeast Glucose Yeast Yeast Yeast
Water (+/-) Water Water Glucose Glucose Water
Conditions
Glucose (+/-) Water Water
Flour (+/-) Flour Heat
Heat (+/-)
Gas in the
balloons
(Yes/No) n/a Yes No Yes No No
Because
yeast must
first break
down
complex Without a
carbs found This
sugar
in flour, combination
fermentation yeast is allows yeast source,
proceeds absent, no to ferment yeast Boiling
more slowly. fermentatio glucose, cannot kills
Explain your Though not n occurs, producing ferment or yeast
observation. as much as and no CO₂ carbon produce which
Why and why with pure dioxide (CO₂) prevent
is produced, gas, so the
glucose, this gas, which
not do you see causes some
leaving the
inflates the
balloon s
the balloon inflation. balloon balloon remains fermen
being inflated? deflated. significantly deflated. tation

Table 2: pH of carbonated water (5 points).

Beaker #1 Beaker #2
Yeast (+/-) Yeast Yeast
Water (+/-) Glucose Flour
Conditions
Glucose (+/-) Water Water
Flour (+/-)
Heat (+/-)
Color of red
litmus paper n/a Red Red

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Color of blue
litmus paper n/a Red Red

Why did the litmus paper change the color? Is the beaker solution acidic, neutral, or basic? (5
points)

The litmus paper changed its color because the beaker is acidic and when red litmus paper is
in acid it stays red when blue litmus paper is in acid it turns red.

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Discussion (5 points per answer):

1. What kind of gas is present in the balloon? Write the equation for this process.

Carbon dioxide

2. Why the balloon wasn’t inflated in the presence of heat?

Because the heat killed the yeast which prevented fermentation

3. Suggest two ways to slow down or stop enzymes used in the respiration process.

• Reduce the Temperature: At lower temperatures, molecular mobility slows down

respiration-related enzymes, which lowers reaction rates. Enzyme activity may potentially
halt in extreme cold.
• Employ an Inhibitor or Denaturation: Enzymes can be inhibited or denatured by
substances like toxins, heavy metals, or extremely high or low pH levels, which changes
their structure and halts respiration. Additionally, boiling denatures enzymes, rendering
them inactive.

4. Explain how cellular respiration is different from fermentation. What are the
differences and similarities between these 2 biological processes? (Use your own words
but don’t forget biological vocabulary.) (4 points)

Cells use both fermentation and cellular respiration to generate energy, although the efficiency
and oxygen needs of each process vary. As an aerobic process, cellular respiration needs oxygen
to fully convert glucose into carbon dioxide, water, and ATP (energy). It generates a large
amount of ATP and takes place in the mitochondria. On the other hand, fermentation does not
require oxygen because it is anaerobic. Fermentation produces far less ATP by converting
glucose into lactic acid in mammals or alcohol and carbon dioxide in yeast and certain bacteria.
Notwithstanding these variations, the first step in both procedures is glycolysis, which releases
energy by breaking down glucose into pyruvate.

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Activity 2: Trouble Brewing
Mr. Barley and Ms. Oat are opening a brewery. Neither of them has brewed beer before, but
they have an instruction manual and the supplies they need, including yeast and barley (which
provides a source of sugar for the yeast). The manual shows the following diagram:

1.

a. What molecule is needed for aerobic respiration but not for fermentation? (2 points)

The molecule needed for aerobic respiration but it fermentation is oxygen. It’s essential to the
Krebs cycle and the electron transport chain where most of the ATP is produced.

b. Which of the two processes yields the most ATP? (2 points)

The two processes that yields the most ATP is aerobic respiration. Fermentation yields 2 ATP
per glucose while aerobic respiration generates up to 29 ATP per each glucose. (2 from Krebs
cycle and up to 25 from electron transport chain.)

Barley and Oat have a problem - the beer they made doesn’t have any alcohol in it! They can’t
figure out why, so they’ve hired you. First, you do an experiment to test whether increasing the
amount of sugar will increase the amount of alcohol produced. The figure shows the results of
your experiment.

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2. Summarize the data in the graph. (6 points)

Your next step was to measure the change in concentrations of O2, CO2 and ethanol.

Time (hours) O2 (%) CO2 (%) Ethanol (%)

0 = beginning 1.00 0.01 0
1 0.99 0.02 0
2 0.98 0.03 0
3 0.97 0.03 0
4 0.97 0.04 0
5 0.96 0.05 0
6 0.95 0.05 0
7 0.94 0.06 0
8 = end 0.94 0.06 0

Now you are ready to put together all the evidence and figure out why the beer doesn’t have
any alcohol and what Barley and Oats should do to make alcoholic beer.

The formula for cellular respiration is C6H12O6 + 6O2 → 6CO2 + 6H2O

The formula for alcoholic fermentation is C6H12O6 → 2C2H5OH + 2CO2

3. Was the yeast in the brewing vats of and Oat’s Brews performing aerobic respiration or
alcoholic fermentation? What evidence supports this conclusion? (4 points)

The yeast was performing aerobic respiration rather than alcoholic fermentation. The oxygen
levels remained consistently stable over time only decreasing slowly. There was also no ethanol
producing it remained at 0%. If fermentation happened it would be a significant increase in
ethanol. Seeing as yeast converts sugar into alcohol and CO2 anaerobically.

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 5. What do you recommend that Barley and Oat do to make their beer contain alcohol? (4
points)

They should remove oxygen from the environment they are making alcohol from, increase its
sugar content, and control temperature to make optimal fermentation conditions.

Humans also use fermentation when oxygen levels are low, but our cells don’t produce alcohol.
5. Identify the molecules (A, B, D-H) and # of ATP produced for each process in the image
below: (20 points)

A: glucose F:pyruvic acid

B: oxygen G:lactic acid
C:32 H:ethanol
D:pyruvate I: 2
E: lactate J: 2
Making Connections:

6. You just started a new exercise routine of running three days a week. On your first run, after
a little while you feel a burning sensation in your calves. (12 points)

1. What’s happening at the cellular level?

Your muscle cells need more energy while you run. They first create ATP through
aerobic respiration. However, your cells switch to lactic acid fermentation when
the oxygen supply is inadequate, such as during vigorous exercise. By
replenishing NAD⁺ and converting pyruvate to lactate, this mechanism permits
glycolysis to proceed and produces some ATP for energy.
2. Why do you think it hurts?

The buildup of lactic acid (or lactate and hydrogen ions) in your muscles is what
causes the burning feeling. This causes the environment to become more acidic
and lowers the pH inside muscle cells, which can disrupt enzyme action and
produce short-term discomfort.

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 3. Why can’t lactic acid fermentation continue indefinitely in your cells?

Because it only generates two ATP per glucose molecule, lactic acid fermentation
is less effective than aerobic respiration, which generates 36–38 ATP.
Furthermore:

A accumulation of lactate can affect muscle function and reduce pH.

Limited NAD⁺ regeneration gradually slows down glycolysis.
Normal ATP generation requires the repayment of oxygen debt.
This explains why you breathe heavily after vigorous activity in order to
replenish oxygen and return to aerobic respiration.