Experiment No.

Alcohol Fermentation

ABSTRACT

Alcohol fermentation is done by yeast and some kinds of bacteria. These microorganisms

convert sugars in ethyl alcohol and carbon dioxide. Yeasts perform this conversion in the

absence of oxygen. Consequently, alcoholic fermentation is considered an anaerobic process. In

this experiment, baker’s yeast is used. In order to effect fermentation, the yeast must possess the

enzyme alcohol dehydrogenase. It catalyzes ethanolic fermentation reaction to form ethanol

using glucose (a carbohydrate) as substrate. Hence we used different classification of sugars in

this experiment to verify its feasibility and distinction in their reaction. Glucose, sucrose,

fructose, glycerol, and starch were tested in this experiment. The rate of production in getting

more precipitate will depend on the sugar component of the sample used. Each of the sugars

have its own fermentation properties.

I. Introduction

All living organisms obtain the energy necessary to sustain life, from the oxidation of

organic substances by molecular oxygen, in the process of respiration. Under anaerobic

conditions, many organisms, including yeast, obtain energy from the process of fermentation. In

alcoholic fermentation, characteristic of many yeast species, the fermentation process starts with

one molecule of the six carbon sugar-glucose, and terminates with two molecules of the two

carbon alcohol-ethanol, and two molecules of CO2.

C6H12O6  2CH3CH2OH + 2CO2

The CO2 released in the process, dissolves in water and forms a carbonic acid. This acid

dissociates to form hydrogen carbonate and hydronium ions:

CO2 + 2H2O  H2CO3  H30+ + HCO3

In acidic solutions, the dissolution of CO2 in water decreases and it is released to the air as CO2

gas.
II. Methodology

A. Equipment
Thermometer
Universal clamp (4) Large test tubes

Corks (4) 25-mL Erlenmeyer flask

Hot plate Iron Stand

Wire Gauze (4) 25 cm glass tubing

Mineral oil Clay

Funnel 250-mL Beaker

Tripod Stirring rod

Beam balance

B. Reagents
Glucose Yeast solution
Sucrose Ba(OH)2
Glycerol Fructose
Starch
Procedure:

Dissolve 4 g of sugar and 0.25 g of disodium hydrogen phosphate which

buffers the mixture to maintain an optimal pH, in 15 mL of warm (40
degrees C) distilled water contained in 50-mL Erlenmeyer flask. Add a
stirbar and 2 g of dry, active baker’s yeast, and assemble the apparatus for
anaerobic fermentation.

The other tube contains a 3% Barium hydroxide solution and should be

protected from atmospheric carbon dioxide by a layer of mineral oil (1
mL). A precipitate of Barium carbonate will form as carbon dioxide is
produced during the course of the fermentation. Stir the mixture
vigorously for 2 h in a warm location to suspend all of the yeast and to
initiate the fermentation.

Dismantle the set up. Remove the mineral oil in the test tube
using medicine dropper.
Prepare the pre-weighed fluted filter paper and filter the
precipitate that is present in the test tube.

Dry the precipitate at 105 degree C for 1 h and weigh.

III. Data and Results

Alcohol Fermentation

Carbohydrate Formation of Gas Amount of Rate of

Precipitate (g) Fermentation
(g precipitate/h)
Glycerol none None none
Glucose 8 minutes 0.512 g 0.256g/h
Sucrose Minimal amount cannot be measured
Fructose 7 minutes 1.301 g 1.301g/h
Starch Minimal amount cannot be measured

IV. Discussion

Glycerol
There was no evident precipitation for glycerol. This is because yeast cannot metabolize glycerol

through the anaerobic fermentation. Glycerol is not a carbohydrate and it cannot be degraded that

is why there is no reaction that occurs in the process.

Glucose
Glucose started its formation of gas in the 8 minute of the experiment. Glucose was observed for

a total of 2 hours which resulted to 0.512 g of precipitate. Glucose produced 0.256 g of

precipitate per hour. Yeast contains enzymes that catalyse the breakdown of glucose to ethanol

and carbon dioxide. Glucose is a monosaccharide sugar and therefore has one unit of sugar. This

will enable the enzymes in the yeast to break down the bonds of the simple sugar very easily

with less energy, and short period of time.

Sucrose
Sucrose resulted with a minimal amount of precipitate which cannot be measured. Sucrose

cannot be fermented directly by the yeast enzyme, zymase. One of yeast's other enzymes,

invertase, must first digest sucrose into glucose and fructose. The yeast enzyme, zymase, then

ferments this sugar.

Sucrose has two units of sugars and therefore has twice as much bonds as glucose and fructose

sugar which will slow down the enzymes’ action in breaking down the bonds, as it requires more

energy with longer period of time to break down the bonds.

Fructose
Fructose produced the greatest amount of by-product in this experiment, yielding 1.301 g of

precipitate per hour. Fructose is fermented faster than glucose by some yeasts. During alcoholic

fermentation, yeasts convert most of the fructose present into alcohol and CO2. Fructose is a

monosaccharide sugar and therefore has one unit of sugar. This will enable the enzymes in the

yeast to break down the bonds of the simple sugar very easily with less energy, and short period

of time.

Starch
Starch produced a minimal amount of precipitate which cannot be measured. Starch is made up

of many glucose units joined together, but yeast cannot digest starch unless it is broken down

into glucose units.

Starch is a polysaccharide. Sugars have to be broken down into simple sugars

(monosaccharides) to allow yeast to react. Therefore, this is the reason why starch produced a

minimal amount of precipitate.

V. Conclusion
VI. Questions and Answers

1. Explain why glycerol is not fermented under the given set of conditions.

From the principle of alcohol fermentation, it converts sugar to ethyl alcohol and carbon

dioxide. In the case of glycerol, which is not a sugar it cannot be degraded that is why there is

no reaction that occurs in the process. But glycerol is the by-product produced on alcohol

fermentation

2. How does yeast get fructose into glycolysis?

The main role of yeast in the process is to convert sugar to ethyl alcohol and carbon dioxide.

Fructose is a sugar, so yeast reacts with the hemiacetal side of the fructose and bring fructose

in the process of glycolysis.

3. What class of glycosidase enzymes must yeast possess to effect fermentation of

carbohydrates?

In order to effect fermentation, the yeast must possess the enzyme alcohol dehydrogenase. It

catalyzes ethanolic fermentation reaction to form ethanol using glucose (a carbohydrate) as

substrate.

4. What monosaccharides would be produced by hydrolysing lactose, maltose, sucrose, and

starch and which of these reactions yeast seem able to carry out? Explain.

The main monosaccharide that would be produced is glucose. Of all these reactions, yeast will

react more with maltose because it contains two glucose units. Glucose is the main sugar that

can react fast to yeast during alcohol fermentation.

5. Write a balanced equation for the reaction of gas with Barium hydroxide.

Barium has a charge of -2 and Oxygen has a charge of +2. Oxygen is nature appears as O2,

therefore Barium + Oxygen = barium oxide; Ba + O2 = BaO; 2Ba + O2 = 2BaO

References:

Essays, UK. (2018). The Process of Fermentation. Retrieved from

https://www.ukessays.com/essays/sciences/fermentation.php?vref=1

Aguilar, P. (2013). Alcohol Fermentation Presentation. Retrieved from

https://www.slideshare.net/PrincessAlenAguilar/alcohol-fermentation-presentation

Bake Info. (n.d). Enzymes. Retrieve from https://www.bakeinfo.co.nz/Facts/Bread-

making/Bread-ingredients/Enzymes

Royal Society of Chemistry. (n.d.). Fermentation of Glucose using Yeast. Retrieved from
https://edu.rsc.org/resources/fermentation-of-glucose-using-yeast/470.article
Mapiscay, Meryll Grace Kristine

Matutino, Julius Adrian

Mendoza, Alleah Johanna

Nario, James Mattew