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INTRODUCTION
organism. It is an emergent property of life that arises from specific interactions between
molecules within the cell. Metabolism is generally concerned with managing the material
and energy resources of the cell. Some pathways of metabolism break down complex
respiration is a major passageway of catabolism, in which the sugar glucose and other
the muscles in your body to contract. These ATP molecules are produced during cellular
use of enzymes, phosphate groups from ATP are being transferred to other compounds,
from one reactant to another. These electron transfers are called oxidation-reduction
reaction, shortly-named redox reactions. During this reaction, the loss of electrons from
one substance is called oxidation, while the addition of electrons to another substance is
First, 6-carbon glucose is broken in half which forms two molecules of pyruvate with
three carbon atoms each. In the second stage, the two pyruvate molecules each lose a
carbon atom by a release of carbon dioxide. In the citric acid cycle, the bonds of the
remaining 2-carbon molecules are broken and the single carbon atoms combine with
oxygen to form more carbon dioxide. Lastly, during oxidative phosphorylation, the
energy from the broken bonds of carbons in glucose is then transferred to ATP (Tobin
However, fermentation provides a mechanism for cells to oxidize organic fuel and
NADPH to either lactate or alcohol and carbon dioxide (Mader, 2001). In lactate
fermentation, the pyruvate is converted into lactase. On the other hand, in alcohol
electrons from NADH and thereby becomes ethanol (Starr and Taggart, 1987). Single-
celled fungi, yeasts, are good examples of organisms that use this pathway and generate
alcohol and carbon dioxide. Yeasts are used to leaven bread. It is metabolically flexible
that it allows them to break down organic molecules through fermentation if oxygen is
absent and through aerobic respiration if oxygen is present. In the absence of oxygen,
yeasts ferment the energy sources and release carbon dioxide gas and ethyl alcohol as by
energy that must be provided by the molecules in order to react is known as the energy of
activation. In laboratories, heat usually supplies the free energy of activation. However,
many different reactions are simultaneously going on in a cell, and heat would cause
harm to all of these reactions. In order to avoid this problem, cells use enzymes. Enzymes
forms a temporary bond with the molecules reacting to lower the activation energy
required for a reaction. Substrate is the molecule on which an enzyme acts. This fits
perfectly on the site of the reactions catalyzed by the enzyme, active site (Raven, et. al.,
2005). Organic compounds such as carbohydrates, fats and proteins can be used as
substrates. However, carbohydrates are essentially used as substrates in yeast (Duka, et.
al., 2009). Carbohydrates are sugars and sugar derivatives and they differ in complexity
The hypothesis being tested in the study is in the statement: If the type of
substrate will affect the rate of cellular respiration, then, the more complex the substrate
is, the slower the rate of respiration. 15 mL of both distilled H2O and 10% yeast were
placed in six Smith fermentation tubes. Each tube was then added by 15mL of the
Within 25 minutes, the height of CO2 gas evolved inside each tube was measured and the
In connection to the hypothesis stated, the study aimed to determine the effect of
the nature of substrate on the rate of respiration. The specific objectives were:
respiration; and,
2. to describe the relationship between the nature of substrate and the rate of
respiration.
The study was conducted in the University of the Philippines Los Baños, Institute
of Biological Sciences, Room C-117 last November17, 2014 under the supervision of Dr.
Blesshe Querijero.
MATERIALS AND METHODS
Smith fermentation tube method was used to determine the effect of the nature of
substrate on the rate of respiration in yeast. Six Smith fermentation tubes were obtained.
Smith fermentation tube is a special tube having a closed vertical arm extending into a
bulbous portion with tapered opening. 15mL of the following solutions were added to the
respective tubes: 1-fructose, 2-glucose, 3-sucrose, 4-lactose, 5-starch and 6-dH2O. All the
solutions were at 10% concentration. Then, 15mL dH2O and 15mL 10% yeast suspension
were added to each tube. It was noted that yeast should be the last to be added to the
tubes. Each tube were gently shaken to make sure that the solutions inside were
thoroughly mixed. Also, trapped bubbles were removed by covering the opening with the
The openings of the tubes were immediately plugged with cotton balls and then
the tubes were tied together at their vertical arms to keep them upright. The tubes were
then set aside in an area where they will not be disturbed. The height of the area occupied
by the CO2 evolved, which were trapped in the vertical arm, were measured every five
The volume of the gas evolved, CO2, was computed using the formula:
𝑉𝐶𝑂2 = 𝜋𝑟 2 ℎ
where: r is the radius of the inner opening of the Smith fermentation tube; and
𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝐶𝑂2
𝑅𝑎𝑡𝑒 𝑜𝑓 𝑅𝑒𝑠𝑝𝑖𝑟𝑎𝑡𝑖𝑜𝑛 = 𝑡𝑖𝑚𝑒 (min)
Table 1 and Figure 1 show the height of carbon dioxide (CO2) bubble formed in
each of the tube after a given period of time. It was observed that after 25 minutes,
fructose had the highest height of CO2 formed of 7.1 cm. this was followed by glucose
with 6.5 cm, sucrose with 3.1 cm and lactose, starch and dH2O having all equal to 0cm.
The tube with dH2O as substrate served as the controlled setup of the experiment. In this
setup, respiration would not occur since there is no organic substance to be broken down
to generate energy. In the tube with starch as substrate, no respiration took place within
cerevisiae must carry out the preliminary steps of breaking down the starches to glucose
first, since yeast cannot directly use these complex carbohydrates (Hopson and
not occur in the setup with lactose as substrate while respiration occurred on sucrose as
substrate. This is due to the absence of the enzyme lactase from the majority of the yeasts
0 0 0 0 0 0 0
In Table 2, the computed volume of CO2 evolved in each setup after 25 minutes
were shown. The tube with glucose as substrate had the highest volume of CO2 evolved
equal to 13.07cm3. It was followed by the tubes with fructose as substrate (12.55cm3) and
with sucrose as substrate (6.233cm3). The tubes with lactose, starch and dH2O as
minutes
Type of
Fructose Glucose Sucrose Lactose Starch dH2O
substrate
VCO2
12.55 13.07 6.233 0 0 0
(cm3)
Using the computed volumes of CO2, the rate of respiration of each tube was
determined as shown in Table 3 and figure 2. The tube with glucose as substrate had the
highest respiration rate equal to 0.52cm3/min. The tube with fructose as substrate
(0.51cm3/min) and the tube with sucrose as substrate (0.25cm3/min) have followed.
Table 3. The respiration rate of yeast with different substrates after 25 minutes
Type of
Fructose Glucose Sucrose Lactose Starch dH2O
Substrate
VCO2
12.55 13.07 6.233 0 0 0
3
(cm )
Rate of
(cm3/min)
Height (cm)
Type of Substrate
Figure 1. A bar graph showing the height of carbon dioxide bubble formed inside the
Type of Substrate
Figure 2. A bar graph showing the respiration rate of yeast with different substrates after
25 minutes.
Fructose and glucose are both monosaccharides that are considered most
important monomers of carbohydrate. They both share the molecular formula C 6H12O6;
however, different properties of these sugars emerge from their different arrangements of
atoms (Hopson and Postlethwait, 1992). Also, fructose and glucose differ in
aldehyde functional group (Mcmurry, 2012). Based from Zubay, et. al. (1995), there is a
faster fermentation when fructose is used than glucose, though they are both
phosphorylate glucose 6-phosphate to fructose 6-phosphate. This would mean that less
time is needed to ferment fructose than glucose; therefore, fructose will have a faster rate
of respiration than glucose. However, the data have shown contradictions to these.
glucose (Hopson and Postlethwait, 1992). Accordingly, sucrose will have a lower rate of
respiration than the aforementioned monosaccharides because it needs to take both of the
Based from the stated data above, the mixture with the complex type of substrate
will result to slower rate of respiration and the mixture with the simple type of substrate
will result to faster rate of respiration. This statement leads to verification of the
forenamed hypothesis, “If the type of substrate will affect the rate of cellular respiration,
then, the more complex the substrate is, the slower the rate of respiration.” However,
random errors possibly occurred during the conduct of the experiment that may have
affected the desirable outcomes to be further discussed in the summary and conclusion.
SUMMARY AND CONCLUSION
The effect of the nature of substrate on the rate of respiration was determined by
the use of the Smith fermentation tube method. Six Smith fermentation tubes were all
placed by 15mL of both dH2O and 10% yeast. Six separate solutions of different types of
substrates were also added to the corresponding tubes: 1-fructose, 2-glucose, 3-sucrose,
4-lactose, 5-starch and 6-dH2O. The height of CO2 bubble formed in each tube within 25
minutes were measured in cm. the gathered data were used to calculate the rate of
respiration of the mixtures in every tube and was presented and analyzed carefully.
Results have shown that the setup with glucose, monosaccharide, as substrate had
the highest rate of respiration (13.07cm3/min) followed by the setup with fructose, a
(6.233cm3/min). It was also observed that the setups with lactose, starch and dH2O as
substrates have no evolution of CO2 gas within the given period of time. Thus, no rate of
the rate of respiration. That means that the higher the complexity of the substrate, the
lower its rate of respiration becomes. However, in the experiment conducted, theoretical
results were not perfectly achieved. In this case, fructose should have a higher respiration
rate than glucose as explained in the results and discussion. The possible error happened
may have been the uncertainties in measurements and usage of devices by the
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