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Determination of the Effect of Different Substrates on the Cellular Respiration in Yeast

using the Smith Fermentation Tube Method1

John Michael Ocampo

Eloise Danise Pantaleon
Rozel Razal
Kim Ruth Renomeron
Group 3 Bio 1 UV-3L

December 1, 2014

A scientific paper submitted in partial fulfillment of the requirements in General Biology 1

laboratory under Prof. Janece Jean A. Polizon, 1st sem., 2014-2015.

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INTRODUCTION
Cellular respiration is the breakdown of organic substances into simple
substances. It is considered a catabolic process wherein energy is released. One type of
cellular respiration needs the presence of oxygen; hence, the name aerobic respiration
(Campbell and Reece, 2011). The breakdown of glucose through cellular respiration is
shown in the equation below:
C6H12O6 + 6H2O + 6O2 6CO2 + 12H2O + energy (456,000 cal/mol)
However, there is another type of cellular respiration that does not require
oxygen which is called anaerobic respiration. It may either produce lactic acid or ethanol.
Energy is also released in this type of respiration; however, the amount is in smaller
quantity compared to the energy yield of aerobic respiration (Duka, Diaz and Villa,
2009). Anaerobic respiration usually takes place in some plants and other
microorganisms. The equation for anaerobic respiration is shown below:
Ethanol: C6H12O6 2CO2 + 2C2H5OH + energy (24,000 cal/mol)
Lactic Acid: C6H12O6 2C3H6O3 + energy (36,000 cal/mol)
Yeast is an example of an organism that undergoes cellular respiration
aerobically and anaerobically. Production of carbon dioxide and ethanol is evident in its
cellular

respiration.

Anaerobic

respiration

and

anaerobic

respiration

occurs

simultaneously in yeast. Yeast, a member of the Kingdom Fungi, is widely used in bread
making and wine making. (Obtaining Energy, n.d.)
In fermentation, yeast generates energy from organic substances in order to fuel
the cellular respiration. These substances are the substrates, which can be proteins,
fats, or carbohydrates. But most of the time, carbohydrates serve as substrates when

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yeast undergoes cellular respiration. Carbohydrates have many variations that range
from simple to complex sugars. Example of simple sugars, which are called
monosaccharides is glucose. Lactose on the other hand is an example of a
disaccharide, meaning two sugars. Carbohydrates that are composed of many sugars,
such as starch, is an example of a polysaccharide. These sugars may vary in some
factors but they are all carbohydrates. This characteristic can be a good factor in
studying the effects of the nature of the substrates on the rate of cellular respiration
(Duka, Diaz and Villa, 2009). In this experiment, the hypothesis will be - with variation in
the nature of substrate to be used in fermentation, no change will be observed in the rate
of cellular respiration in yeast.
This study aims to observe the cellular respiration in yeast using the Smith
fermentation tube method and to identify the effect of using different substrates in the
cellular respiration of yeast.
Specifically, the objectives are:
1. to differentiate the cellular respiration in yeast, both aerobically and
anaerobically,
2. to measure the rate of cellular respiration in yeast by observing the CO 2
evolution in fermentation; and
3. to recognize the relevance of pressure as an indication of gas accumulation
(CO2 evolution) in the Smith fermentation tube.
The experiment will be conducted on November 24, 2014 at Room C-127,
Institute of Biological Sciences, University of the Philippines Los Baos, Laguna.

MATERIALS AND METHOD

Cellular respiration in Brewers yeast was tested using the Smith fermentation
tube method. First, four Smith fermentation tubes were obtained (Figure 1).

Figure 1. Smith fermentation method.

The tubes were labeled A, B, C and D respectively. Markings with 1 cm interval
were placed on the vertical arm portion of each tube.
*insert substrate preparation*
Next, the prepared 15 mL solutions of the different substrates were poured on
their respective tubes: A sucrose, B glucose, C fructose and D distilled water.
Tube D served as the control group in the experiment. Afterwards, 15 mL distilled water
was transferred to each tube.
*insert yeast preparation*
Next, the prepared 15 mL solutions of yeast were poured on their respective
tubes
The order of pouring of substances into the tubes was strictly followed.

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The mixture was shaken gently. No bubbles should appear on the tubes before
the set-up is complete. If in case bubbles will be trapped on the closed end of the tube,
cover the tubes with the palm of one hand and tilt the tubes horizontally. This will allow
the bubbles to escape from the tube.
The openings of the tubes were plugged with cotton balls. The tubes were tightly
sealed to make sure that air will not be able to enter the tube once cellular respiration
has started. The tubes were tied together immediately and placed in an upright position.
Avoid holding the tubes for a long time because the heat generated by hands will have
an effect on the respiration of yeast. After a short period, observe the production of
bubbles on the vertical arm of the tubes. This means that the mixture has undergone
CO2 evolution, a proof of the occurrence of cellular respiration. After bubbles have
formed, measure the height of the area occupied by CO2 evolved every five minutes for
30 minutes. Then, compute for the volume of gas evolved using:

V CO2 evolved = r 2 h
where: r = radius of the Smith fermentation tube
h = height occupied by gas evolved

The rate of CO2 evolution (in cm/min) for different substrates will also be
calculated. The results will be tabulated and observed.

Schematic Diagram of Smith Fermentation Tube Method

substrate (sucrose, glucose,

fructose) or dH2O
-

pour 15 mL 10% substrate or dH 2O

into Smith fermentation tube
add 15 mL dH2O

substrate solution +
dH2O
-

add 15 mL 10% yeast suspension

substrate solution +
dH2O + yeast
-

shake mixture gently

remove trapped bubbles from closed
end
plug cotton balls on open end
place in an upright postion

substrate solution +
dH2O + yeast
(covered)
-

(no) CO2 gas evolution

bubbles will (not) be formed

fermented yeast
-

Measure height occupied by CO2 (if

any)
Calculate for volume of gas and rate
of CO2 evolution

RESULTS AND DISCUSSION

Table 1. Height occupied by area of CO2 evolved

Time (minutes)
0
5
10
15
20
25
30
Final height

A (sucrose)

Height (cm)
B (glucose)
C (fructose)

D (dH2O)

Table 2. Volume of gas evolved and rate of CO2 evolution

A (sucrose)
3

Volume (cm )
Rate (cm/min)
Expected Rank
Actual Rank

B (glucose)

Set-ups
C (fructose)

D (dH2O)

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12
10
8

Rate of CO2 evolution (cm/min)

6
4
2
0

Substrate

Figure 2. Comparison of the rate of CO2 evolution of different substrates.

LITERATURE CITED

Campbell, N.A., Reece, J.B., et.al. (2008). Biology (8th ed). San Francisco, CA: Pearson
Education Inc., Benjamin Cummings.

Duka, I.A., Diaz, M.G.Q., Villa N.O. (2009). Biology 1 Laboratory Manual: An
Investigative Approach (9th ed). College, Laguna. Philippines: University of
the Philippines Los Baos. pp. 51-54.

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Obtaining Energy (n.d). BBC-GCSE: Useful Products from Respiration (internet article).
Retrieved on November 20, 2014 from:
http://www.bbc.co.uk/schools/gcsebitesize/science/add_ocr_21c/life_processes/e
nergyrev5.shtml