Patrick McCrystal

Cellular Respiration: From O2 to CO2

Purpose:
This lab provided insight to the process of cellular respiration and how it is affected by
temperature in both germinating and dormant pea seeds. Cellular respiration is an ATP-producing
catabolic process in which the electron receiver is an inorganic molecule. It is the release of
energy from organic compounds by chemical oxidation in the mitochondria within each cell.
Carbohydrates, proteins, and fats can all be metabolized, but cellular respiration usually involves
glucose: C6H12O6 + 6O2 → 6CO2 + 6H2O + 686 Kcal of energy/mole of glucose oxidized.
Cellular respiration involves glycolysis, the Krebs cycle, and the electron transport chain.
Glycolysis is a catabolic pathway that occurs in the cytosol and partially oxidizes glucose into
two pyruvate (3-C). The Krebs cycle occurs in the mitochondria and breaks down a pyruvate
(Acetyl-CoA) into carbon dioxide. These two cycles both produce a small amount of ATP by
substrate-level phosphorylation and NADH by transferring electrons from substrate to NAD+.
The Krebs cycle also produces FADH2 by transferring electrons to FAD. The electron transport
chain is located at the inner membrane of the mitochondria and accepts energized electrons from
enzymes that are collected during glycolysis and the Krebs cycle, and couples this exergonic
slide of electrons to ATP synthesis or oxidative phosphorylation. This process produces most of
the ATP. Cellular respiration can be measured in two ways: the consumption of O2 (how many
moles of O2 are consumed in cellular respiration) and production of CO2 (how many moles of
CO2 are produced in cellular respiration). PV = nRT is the formula for the inert gas law, where P
is the pressure of the gas, V is the volume of the gas, n is the number of molecules of gas, R is
the gas constant, and T is the temperature of the gas in degrees K. This law shows several
important things about gases. If temperature and pressure are kept constant then the volume of
the gas is directly proportional to the number of molecules of the gas. If the temperature and
volume remain constant, then the pressure of the gas changes in direct proportion to the number
of molecules of gas. If the number of gas molecules and the temperature remain constant, then
the pressure is inversely proportional to the volume. If the temperature changes and the number
of gas molecules is kept constant, then either pressure or volume or both will change in direct
proportion to the temperature.

Methods:
During the lab, we prepared both a room temperature and a 10oC water bath. Then, we
filled a 50 mL graduated cylinder halfway with water. We added 25 germinating peas and
determined the amount of water that was displaced. Then we removed the peas, placed them on a
paper towel, refilled the graduated cylinder, and added glass beads to the graduated cylinder until
the volume was equivalent to that of the expanded germinating peas. We removed the beads,
refilled the graduated cylinder, added 25 non-germinating peas, and then added more glass beads
until the volume was once again equal to the germinating peas’ volume. After all that was done,
we prepared another set of peas and beads for the last 3 respirometers. Assembly of the
respirometers was the next step. We obtained 6 vials, stoppers, and graduated pipettes. Then we
placed a wad of absorbent cotton in the bottom of each vial and, using a pipette, saturated the
cotton with about 2-3 mL of 15 % KOH. We then placed a layer of non-absorbent cotton on top
of the KOH-soaked cotton in order to protect the peas from the KOH. We placed the first set of
germinating peas, dry peas and beads, and beads alone in vials 1, 2, and 3, and the second set in
vials 4, 5, and 6, then placed the stoppers in each vial. We made slings out of masking tape in
 order to hold the pipettes out of the water for the 10 minute equilibration period and placed the
vials on them (1, 2, and 3 in the room temperature bath, 4, 5, and 6 in the 10oC bath). The 10
minute period was necessary to ensure that a difference in temperature between the air in the vial
and the water would not skew our results. Once the vials were properly adjusted, we lowered
them into the water. Thankfully, the water did not rush into the respirometer, which would have
indicated a leak. We then recorded the reading on the pipette at set time periods.

Results:

Measurement of O2 Consumption by Soaked and Dry Pea Seeds at Room Temperature and
10˚C

Actual Beads Alone Germinating Peas Dry Peas and Beads

Time
Temp Reading Reading Corrected Reading Corrected
(Min) Diff. Diff. Diff.
(oC) at time X at time X Diff.∆ at time X Diff.∆

Initial-
21 .87 .9 .89
0

0-5 20 .9 -.03 .79 .11 .14 .9 -.01 .02

0-10 20 .9 -.03 .7 .2 .23 .89 0 .03

0-15 20 .87 0 .62 .28 .28 .89 0 0

-20 20 .86 .01 .5 .4 .49 .89 0 -.01

Initial-
9 .86 .85 .89
0

0-5 Oxygen
10 Comsumption
.84 .02 .79 of.06
Germinating
.04 Peas,.02Non -0
.87

0-10 Germinating
10 .85 Peas,
.01 and Glass
.74 .11 Beads
.1 at.9 20˚C
-.01and -.02

0-15 10 .87 .01 .69 10˚C

.16 .17 .92 -.03 -.02
Beads Alone 20˚
0-20 10 .89 -.03 .65 .2 .23 .95 -.06 -.03
1
Germinating Peas
0.8 20˚
Consumption

Dry Peas and Beads

Oxygen

0.6
20˚
0.4
Beads Alone 10˚
0.2
0 Germinating Peas
0 0-5 0-10 0-15 -20 10˚
Time Dry Peas and Beads
10˚
 It was necessary to compare the reading from the peas with the reading from the
beads because the beads served as a control variable, therefore, the beads experienced no
change in gas volume. Germinating seeds have a higher metabolic rate and need more
oxygen for growth and survival. Non-germinating peas, though alive, need to consume
far less oxygen in order to survive. The KOH absorbed the carbon dioxide and caused it
to form a precipitate at the bottom of the vial, preventing it from changing the pressure in
the vial. When the peas underwent cellular respiration, they consumed oxygen and
released carbon dioxide, which reacted with the KOH in the vial, resulting in a decrease
of gas in the pipette. The water moved into the pipette because the pressure in the pipette
lessened.

Calculations:

Condition Calculations Rate in mL O2/ minute

(0.85-0.65)
Germinating Peas/ 10 oC .01
20 min.

(0.9-0.5)
Germinating Peas/ 20 oC .02
20 min.

(0.89-0.95)
Dry Peas/ 10 oC -.003
20 min.

(0.89-0.89)
Dry Peas/ 20 oC 0
20 min.
Conclusion:
The lab demonstrated many important things relating to cellular respiration. It showed
that the rates of cellular respiration are greater in germinating peas than in non-germinating peas.
It also showed that temperature and respiration rates are directly proportional; as temperature
increases, respiration rates increase as well. Because of this fact, the respirometers placed in the
water at 10 oC displayed a lower rate of cellular respiration than the respirometers placed in the
room temperature water. The non-germinating peas consumed far less oxygen than the
germinating peas. This is because, though germinating and non-germinating peas are both alive,
germinating peas require a larger amount of oxygen to be consumed so that the seed will
continue to grow and survive. In the lab, CO2 made during cellular respiration was removed by
the potassium hydroxide (KOH) and created potassium carbonate (K2CO3). It was necessary that
the carbon dioxide be removed so that the change in the volume of gas in the respirometer was
directly proportional to the amount of oxygen that was consumed. The result was a decrease in
gas volume within the tube, and a related decrease in pressure in the tube. The respirometer with
just the glass beads served as a control group that did not undergo cellular respiration. Numerous
errors could have occurred throughout the lab. The temperature of the baths may have been
allowed to fluctuate, which would change the temperature in the vials. The amounts of peas,
beads, KOH, and cotton may have varied from vial to vial. Air may have been allowed to creep
into the vial via a leaky stopper or poorly sealed pipette. The vials may have not properly
equilibrated, and students could have read the pipettes either too soon or too late. Students may
have misread pipettes. KOH could have come into contact with the sides of the vials when it was
being dropped onto the cotton. Mathematical inaccuracies may have occurred when completing
the table.