On the Rate of Yeast Glycolysis in Anaerobic Respiration and the Usefulness of Varied

Carbohydrate Nutrients and the Effect of Temperature (revised)

Kalvin Foo

Dept. of Biology, TCNJ

Dr. Kress

December 8, 2009
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Abstract:

As yeast (Saccharomyces cerevisiae), is placed within an anaerobic environment,

baseline ATP production is dependent on glycolysis, which is sustained via alcoholic
fermentation. Evaluated was the general respiration rate of yeast, but the secondary purposes
were to compare the ability of the organism to metabolize the sugars glucose, galactose, sucrose,
lactose, maltose, and fructose and also to observe the difference in respiration rates as per varied
temperature. By measuring the production of CO2, the rate of glycolysis performed by yeast can
be measured via that byproduct, which is directly correlated to its catalysis of sugar into energy.
After testing these sugars in 20oC, 30oC, and 40oC, it was determined that simpler sugars, such as
glucose, fructose, and carbohydrates composed of those two (sucrose and maltose) are able to be
utilized by yeast for glycolysis while complex sugars (galactose and lactose) had little value in
terms of anaerobic respiration (fig. 2). Finally, an average the Q10 of 2.27 was calculated for
sucrose and glucose, showing that the respiratory rate of yeast roughly doubles with every
increase of 10oC (fig. 3).

Introduction:

Anaerobic respiration, as an alternative ATP producing process, is the process by which

fermentation replenishes energy-carrying molecules such as NAD+ to maintain a cycle.

Compared to normal cellular respiration, which include ATP replenishment through the Citric

Acid Cycle and oxidative, ADP phosphorylation in both aerobic and anaerobic respiration

include glycolysis. Eukaryotic organisms, such as the model organism yeast, make use of

anaerobic respiration when the situation prevents enough oxygen to reach a cell, such as in

strenuous muscle activity in a multicellular organism or an oxygen poor environment for smaller

organism such as yeast (Van Waarde, 1991). These cycles each require a fermentation step,

either requiring lactic acid or ethanol fermentation, and yeast compliment sugar breakdown with

ethanol formation.

The model organism yeast (Saccharomyces cerevisiae) is a eukaryotic specimen that can

undergo alcoholic fermentation, which is widely used in baking or beer-brewing. In an anaerobic

environment it ferments sugars within the cytosol, producing 2 ATP in the reaction
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C6H12O6 → 2 C2H5OH + 2CO2 + energy (Lovett and Shevlin, 2009).

Because ethanol is clear, it cannot be measured via spectrophotometery, and energy cannot be

quantified easily. Thus, the production of CO2 can be measured with a respirometer (fig. 1) and

glycolytic rates can be observed.

Two variables were introduced into the reaction were complex sugars and temperature.

The sugar breakdown is facilitated by specific enzymes, such as the usage of zymase for glucose

breakdown, but the lab further evaluated the metabolic usefulness of the carbohydrate nutrients

galactose, sucrose, lactose, maltose, and fructose along with glucose (Bisson and Fraenkel,

1983). Also, tests were run at different temperatures to assess the respiration rate of yeast in

order to determine a Q10 value, a value that shows the change of rate of respiration in yeast at

a difference of ten degrees Celcius.

Materials and Methods:

An integral part of yeast tests is to use samples from the same suspension, so all tests

were performed from the same yeast solution. Samples were incubated in testing conditions

without sugar originally to burn off residual sugars, and then were incubated for 41 minutes with

the specific sugar solutions. For every condition, yeast was tested with a .1M solution of the

sugars glucose, galactose, sucrose, lactose, maltose, and fructose. Each sugar-yeast respirometer

setup was repeated within three temperature settings of 20oC, 30oC, and 40oC, and the measured

displacement in the yeast tube was used to calculate moles of CO2 produced, and then the rate.

These results were evaluated against a negative control of purely a yeast suspension to determine

a 95% percent confidence interval and significant data values where N= 3 as well as a Q10 value.

Suspension preparation and calculations were performed as per Lab manual (Lovett and Shevlin,

2009).
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Results:

After evaluating the values presented by different tests on yeast fermentation, the sugars

that are metabolically significant were determined to be glucose, sucrose, fructose, and maltose

(fig. 2). An example of this is the rate of change in moles/hour in the 40 degree setup, where the

rates were as followed: glucose- 55.4 mmol/hour, galactose- 0 mmol/hour, sucrose- 88.2

mmol/hour, lactose- 1.3 mmol/hour, maltose- 87.3 mmol/hour, and fructose-46.5 mmol/hour,

showing lactose and galactose usage to read significantly minimal due to the 95% confidence

interval where p< .05. Also, there is a significant rise in CO2 production rates, thus glycolytic

rates with an increase in temperature, as also displayed by the values in Fig. 2. Data was

collected from six groups, each performing tests with each sugar in two temperature conditions,

but data collected fell within the 95% confidence interval. The evaluation of rate was performed

alongside a negative control set with just the yeast suspension, the difference of which was used

to adjust rate calculations. Q10 value calculations were performed between 20 and 30oC with

glucose and sucrose, yielding values 1.98 for glucose and 1.87 for sucrose respiration (fig.3).

Discussion:

Results in this lab yielded distinct conclusions regarding the ability for yeast

(Saccharomyces cerevisiae) to metabolize simple and complex sugars. Because the organism

uses enzymes to initiate the sugars into glycolysis, it makes sense that the enzymes are able to

catalyze specific sugars. The setups that showed glucose, fructose, maltose, and sucrose were the

best metabolized by yeast while there was difficulty breaking down galactose and lactose. From

this, the difficulty in processing galactose and lactose derives from the necessity to isomerize the

galactose molecule in both (lactose is a disaccharide of galactose and glucose), and this also
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explains the minimal respiration shown in the lactose setup. On the other hand, maltose is a

disaccharide of two glucose molecules and sucrose is a disaccharide of a glucose and fructose

molecule, both able to be utilized by the yeast’s glycolytic functions.

Next, since the process is partially conducted with enzymes, temperature changes affect

the rate by which CO2 is produced. However, more important is the fact that yeast is a eukaryotic

organism, and eukaryotic organisms have faster metabolisms at higher temperatures, thus faster

respiration. In accordance to this, the Q10 value is about 2, which is normal for most plant and

fungi because yeast classified as a fungus. It is likely this relates to the life cycle of a fungus, as

they thrive in warmer temperatures, so naturally metabolic functions would increase at a warmer

temperature.
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Fig. 1- Respirometer setup, prepared as per Lab manual (Lovett and Shevlin, 2009).

Fig. 2- Bar graph comparing the Glycolytic rates of yeast when in the presence of different
sugars and also varied temperatures, 95% confidence displayed with sample size N=3

Temp Q10 Values for Glucose and Sucrose

Diff
20 – 30 1.98, 1.87
30 – 40 2.58, 2.64
Q10 average- 2.27
Fig. 3- Table containing calculated values of Q10 using equation (R2/R1)(10/(T2-T1)) where R is rate of
CO2 production and T is Temperature in Kelvin.

Works Referenced:

Bisson, Linda F., and Dan G. Fraenkel. "Involvement of Kinases in Glucose and Fructose Uptake
by Saccharomyces cerevisiae." Proceedings of the National Academy of Sciences of the
United States of America 80.6 (1983): 1780-783. JSTOR. 1 Nov. 2009.
<http://www.jstor.org>.

Lovett, D.L., and D.E. Shevlin. Laboratory Manual for Biology 185: Themes in Biology. Ewing:
The College of New Jersey: Department of Biology, 2009. 5-1-5-10.

Van Waarde, Aren. "Alcoholic Fermentation in Multicellular Organisms." Physiological

Zoology 64.4 (1991): 895-920. JSTOR. 1 Nov. 2009. <http://www.jstor.org>.