BIOLOGY

INVESTIGATORY PROJECT

Submitted for ANNUAL

EXAMINATION

2022- 2023

YEAST REPRODUCTION IN SUGAR

SUBSTITUTES

Done by

BHAVADARSHINI.R.
G

XII C

SCIENCE STREAM

SUBMITTED TO DEPARTMENT OF
BIOLOGY
BONAFIDE
 ACKNOWLEDGEMENT

I would like to express my special thanks of gratitude to my teacher Mrs.

Gemcy Ebenezer for her guidance, support throughout the duration of the
project. We completed the project successfully by her motivation and her
extended support for us.

As well as I would like to thank our Correspondent Dr. R.Kishore

kumar , our Principal Mrs. Shanthi Samuel and Vice Principal Mrs. Angelin
Christopher Roy who gave me the opportunity to work on this project, which
in turn helped me in doing a lot of Research and analysis in the chosen topic.

Finally, I would also like to thank my parents and friends who

supported me in completing this project within the limited time frame
 TABLE OF CONTENTS

S.No Content Pages

1 Abstract 01

2 Introduction 02

3 03
Industrial uses of yeast – brewing and

distilling

4 Yeast 04

5 Experiment 06

6 The effect of sugar in yeast 11

7 Fermentation of sugar in yeast 13

8 Uses of Yeast 16

9 Production of Yeast 17

10 Conclusion 18

11 Bibliography 19
 ABSTRACT

There's nothing quite like the smell of fresh-baked bread to make your
mouth water! As any baker can tell you, you can't bake bread without
yeast. This project makes clever use of bread dough to measure yeast
reproduction three different ways, and investigates how well yeast
grow with sugar substitutes as a food source. The purpose of this
project is to see if yeast will reproduce using various sugar
substitutes.

Yeast is a single-cell organism, called Saccharomyces cerevisiae,

which needs food, warmth, and moisture to thrive. It converts its
food—sugar and starch—through fermentation, into carbon dioxide
and alcohol. It's the carbon dioxide that makes baked goods rise. Yeast
is an egg-shaped single-cell fungus that is only visible with a
microscope. It takes 20,000,000,000 (twenty billion) yeast cells to
weigh one gram. To grow, yeast cells digest food and this allows them
to obtain energy.
 INTRODUCTION

Did you ever wonder how bread gets its "spongy" structure? If you've
ever baked homemade bread yourself, you know that you need yeast
to make the bread dough rise. Yeasts are single-celled fungi. Like the
cells in your body, they can derive energy from sugar molecules.
They can also break down larger carbohydrate molecules (like
starches present in flour) into simple sugar molecules, which are then
processed further. Yeast can extract more energy from sugar when
oxygen is present in their environment. In the absence of oxygen,
yeast switch to a process called fermentation.

With fermentation, yeast can still get energy from sugar, but less
energy is derived from each sugar molecule. In addition to deriving
less energy with fermentation, the end products of sugar metabolism
are also different. When oxygen is present, the sugar molecules are
broken down into carbon dioxide and water (plus energy that the yeast
uses to grow and reproduce). In the absence of oxygen, the sugar
molecules are not broken down completely.

The end products are alcohol (with two carbon atoms) carbon dioxide
(one carbon atom), and water. Less energy is extracted from each
sugar molecule: the energy that could be extracted from the alcohol
molecule if oxygen were present. As you know, carbon dioxide is a
gas (at least at room temperature and atmospheric pressure, for you
gas law aficionados). In bread dough, carbon dioxide produced by
yeast forms bubbles that make the dough rise, and give bread its
spongy texture.
 INDUSTRIAL USES OF YEAST – BREWING AND
DISTILLING

Brewer’s and distiller’s yeast strains are unicellular cells belonging to

the genus Saccharomyces. Their functional objectives are to
consistently metabolise wort constituents into ethanol, carbon dioxide
and other fermentable products in order to produce beer – and
potentially spirits – with satisfactory quality, drinkability and
stability. Additionally, brewer’s yeast cultures should be confidently
collected and reused in a subsequent wort fermentation.

Yeasts are unicellular fungi and consequently are eukaryotes1. The

yeast species that has been most closely associated with humankind is
Saccharomyces cerevisiae (Figure 1, page 3) and has long been used
for brewing, distilling (for both potable and industrial alcohol),
winemaking, and baking bread. It is also employed for yeast extracts
in food and flavouring, as well as for therapeutic purposes. It is by far
the most studied and best understood species of the yeast domain, and
is an important model system for basic research into the biology of
eukaryotic cells in general! Indeed, the ability to rationally manipulate
all aspects of its gene expression by in vitro genetic techniques offers
S. cerevisiae a unique place amongst eukaryotes. A eukaryote is any
organism whose cells contain a nucleus and other organelles enclosed
within membranes (for example, animals, plants and fungi; not
bacteria which are prokaryotes) and a plasma membrane, together
with a cell wall surrounding the whole cell.

INTEREST IN THIS DIFFUSE GROUPING OF FUNGI CAN BE

DIVIDED INTO THREE AREAS
1. As an experimental eukaryote
2. A few yeast genomes (not Saccharomyces) contain species (for
example, Candida albicans) that are animal (including human)
pathogens that cause disease (for example, candidiasis)
3. As already described, yeast (particularly Saccharomyces) has
significant industrial and economic value – ethanol production,
etc.

YEAST
Over the years, the taxonomy of S. cerevisiae has undergone major
changes, especially with the impact of molecular biology on its
classification. In 1970 there were 41 Saccharomyces species
identified. However, since then extensive species and genome
rearrangements have been proposed. In 1990 seven species were
agreed with a number of subspecies and varieties. Currently, some
taxonomists refrain from differentiating between closely related
Saccharomyces solely on the basis of DNA-DNA hybridisation and
place these species in S. cerevisiae (Figure 2)3.

THE INDUSTRIAL USES OF YEAST CAN BE DIVIDED

INTO SIX CATEGORIES:
1. Potable ethanol – beer, cider, wine and spirits (whisky, gin,
vodka, brandy, rum, liquors, etc.)
2. Industrial ethanol – fuel, pharmaceuticals, sterilants and solvents
3. Baker’s yeast, biomass (human and animal feeds), flavouring and
carbon dioxide
4. Yeast extracts – cell walls, membranes, mannans, glucans,
vitamins and food flavourings
5. Heterologous proteins and peptides (for example, lipase, thermo-
stable proteinases, phenol oxidase, cellobiase and thaumatin)
6. A plethora of medicinal applications (for example, insulin,
interferon, vitamin supplements).
The significance of this yeast as a fermentative species, particularly
for its role in alcoholic fermentations, has urged many scientists to
study the factors governing the growth, survival and biological
activities of this critical species in different food ecosystems. Strains
of S. cerevisiae are able to ferment hexose sugars, such as D-glucose,
D-fructose, D-mannose and D-galactose. Other larger sugars that can
be fermented by most strains include sucrose, maltose and
maltotriose. Wort is unfermented beer and whisky, prior to
fermentation and distillation. One of the major advances in brewing
science, over the past 40 years or so, has been the elucidation of the
mechanisms by which a yeast cell utilises, in an orderly manner, a
plethora of wort nutrients.

EXPERIMENT
CONCEPTS TO LEARN

Yeast
Fungus (plural: fungi)
Sugar
Carbohydrates
Fermentation
Yeast metabolism
Carbon dioxide
MATERIALS REQUIRED:

• 250-mL graduated cylinder

• 100-mL graduated cylinder
• Wide-mouth, 8 oz. squirt bottles (4)
• Clear plastic tubing
• Waterproof thermometer
• You will also need to gather these items:
• Dry yeast..
• Plastic tub or bucket
• Water
• Optional: Plastic wrap
• Packing tape
• Permanent marker
• Measuring spoons
• Sugar (3 tsp.)
• Sugar substitutes (3 tsp. of each type),
• Saccharin
• Sucralose
• Aspartame. The commercial name is NutraSweet.
• Acesulfame potassium, also known as Ace-K
• Measuring cup
• Warm water, typically 43-46°C (about 110°F–115°F), but
consult the recommendations on your yeast package
• Clock or timer
• Lab notebook
WORKING PROCEDURE
SETTING UP:
1. Remove the small red cap from one of the squeeze bottles. Then
connect the tubing to the tip opening,. Make sure that you have a
tight fit.
2. You will be collecting carbon dioxide from the yeast by displacing
water trapped in an inverted graduated cylinder. Here's how to set
it up:
Fill your plastic dishpan (or bucket) about one-third full with
water.
Fill the 100-mL graduated cylinder with water.
3.The graduated cylinder should now be upside down, full of water
and with its opening under the surface of the water in the dishpan.
Place the free end of the tubing from the plastic bottle inside the
graduated cylinder. Your apparatus is now ready to trap carbon
dioxide from the yeast
4.You can test your gas collection apparatus by removing the tube
from the bottle top and blowing gently into the tube. The bubbles you
create should be captured inside the cylinder
EXPERIMENTAL PROCEDURE

1. Using a permanent marker, label each of the bottles with the type
of solution you will be feeding the yeast (e.g., sugar, nothing,
saccharin, sucralose, aspartame, acesulfame potassium). If you
need more than four bottles, you can re-use them. Make sure to
rinse them out thoroughly between experiments.
2. Dissolve 1 teaspoon (tsp.) of sugar in ½ cup of warm water
(110°F–115°F). When the sugar is fully dissolved, add ½
teaspoon of yeast, mix and pour into the appropriate bottle. Be
sure to note the actual temperature of the water in your lab
notebook.Cap the bottle tightly with your "tube cap," and
place the open end of the tube inside your gas collecting
cylinder. Notethe starting time in your lab notebook.
3. Within 5–10 minutes, the yeast solution should start foaming, and
you should see bubbles collecting in the graduated cylinder. Note
the time when you first start seeing bubbles in your lab notebook.
4. Decide how long to collect CO2 (somewhere between 15–30
minutes is probably good, but you may need to adjust for your
particular conditions). Use the same amount of time for all of your
tests.
5. When the time is up, note how much CO2 was collected by
observing how much water was displaced from the graduated
cylinder.
6. Re-fill your gas collection cylinder, and carefully rinse out the
yeast solution from the bottle. You should run at least three
separate trials for each food source.
7. For each of the sugar substitutes, use the properly labeled bottle.
When preparing your yeast solution, use the same temperature for
the warm water and the same amount of yeast (½ tsp.). Use 1 tsp.
of each sugar substitute instead of sugar.
 PREPARATION AT HOME
THE EFFECT OF SUGAR IN YEAST
 OBSERVATION:

When yeasts eat sugar and turn it into energy, they also produce
carbon dioxide. This process is known as fermentation.
In this activity, the balloons on the bottles should have captured
carbon dioxide produced by the yeasts during fermentation. In the
bottle that contained yeasts but not sugar, the yeasts did not have food
(i.e., sugar) so the balloon should not have inflated. In the bottle that
contained yeasts and sugar (but not salt, baking soda or vinegar), the
yeasts should have thrived and made a lot of carbon dioxide, clearly
inflating the balloon. When salt, baking soda or vinegar was added,
the yeasts should have made less carbon dioxide, inflating the balloon
less than when only sugar was used. This is because the addition of
these substances changed the environment and made it less ideal for
the yeasts. Specifically, adding salt increased the salinity of the
environment, and adding baking soda or vinegar changed the pH of
the environment, making it more basic or acidic, respectively,
compared to the neutral environment provided by the plain water.

EXPERIMENT:
FERMENTATION OF YEAST IN SUGAR
 Procedure

• Fill the medium-sized pot or bowl with at least eight cups of very
warm tap water. Adjust the temperature of the hot water coming from
the tap until it is almost too hot to hold your hands under. Use this
temperature of water to fill the pot.
• Using the warm water from the pot, fill each bottle with about two
and one-half cups (or about one-third full). Put the lid back on to each
bottle and shake them each thoroughly to dissolve all of the
ingredients.
• To each bottle, add two packets of dry yeast (or an equivalent amount
from a jar). Put the lid back on to each bottle and shake each one
gently to mix in the yeast.
• Remove each lid and stretch a balloon completely over the opening of
the bottle (over all of the ridges). Why do you think it is important to
form a tight seal with the balloon on the bottle’s opening?
• Leave the bottles to rest in a warm location for 45 minutes. Keep the
balloons out of direct sunlight. How do the balloons change over
time?
• After 45 minutes, examine the bottles and the balloons. Which
balloons have become inflated? How big are they compared to each
other? Do you notice any differences in the contents of the bottles?
• In which environment did the yeast make the most carbon dioxide?
What does this tell you about the conditions needed for yeast
fermentation to take place?

OBSERVATION

When yeasts eat sugar and turn it into energy, they also produce
carbon dioxide. This process is known as fermentation. In this
activity, the balloons on the bottles should have captured carbon
dioxide produced by the yeasts during fermentation. In the bottle that
contained yeasts but not sugar, the yeasts did not have food (i.e.,
sugar) so the balloon should not have inflated. In the bottle that
contained yeasts and sugar (but not salt, baking soda or vinegar), the
yeasts should have thrived and made a lot of carbon dioxide, clearly
inflating the balloon. When salt, baking soda or vinegar was added,
the yeasts should have made less carbon dioxide, inflating the balloon
less than when only sugar was used. This is because the addition of
these substances changed the environment and made it less ideal for
the yeasts. Specifically, adding salt increased the salinity of the
environment, and adding baking soda or vinegar changed the pH of
the environment, making it more basic or acidic, respectively,
compared to the neutral environment provided by the plain water.
USES OF YEAST
Yeasts have two main uses in food production: baking and making
alcoholic beverages. They have been used in this way since ancient
times – there is evidence that ancient Egyptians used yeast in bread
making, and we have been making fermented drinks like beer and
wine for millennia.

Baking

Baked goods like bread rise because of the presence of yeast as a

raising, or leavening, agent. The most common yeast used in bread
making is Saccharomyces cerevisiae. It feeds on the sugars present in
the bread dough, producing the gas carbon dioxide. This forms
bubbles within the dough, causing it to expand. Other ingredients in
the mixture have an effect on the speed of the fermentation – sugar
and eggs speed it up; fats and salt slow it down.

Winemaking

The alcohol in wine is formed by the fermentation of the sugars in

grape juice, with carbon dioxide as a byproduct. Yeast is naturally
present on grape skins, and this alone can be sufficient for the
fermentation of sugars to alcohol to occur. A pure yeast culture, most
often Saccharomyces cerevisiae, is usually added to ensure the
fermentation is reliable. Sparkling wine is made by adding further
yeast to the wine when it is bottled. The carbon dioxide formed in this
second fermentation is trapped as bubbles.
PRODUCTION OF YEASTS

Ripening yeasts, like lactic acid bacteria, are also produced in

specialised fermentors under strict hygiene conditions but with
different nutrient sources and growth parameters. Notably, yeast
fermentation needs to be conducted under aerobic (with oxygen or
air) conditions. They are concentrated and mainly available as freeze-
dried powders.

The baker’s yeast is commercially produced on a nutrient source

which is rich in sugar (usually molasses: by product of the sugar
refining). The fermentation is conducted in large tanks. Once the
yeast fills the tank, it is harvested by centrifugation, giving an off-
white liquid known as cream yeast. This is further processed into any
of several different forms:

• Compressed yeast: still widely used commercially, it is a soft

beige solid block with limited storage properties.

• Active dry yeast: dried yeast presented in granules or beads that

needs to be rehydrated before it can be used.

• Instant yeast: vacuum packed fine powder that has become

popular in home breadmaking, as it is easy to use
CONCLUSION:

The results show that while sucrose readily undergoes mass loss
and thus fermentation, lactose does not. Clearly the enzymes in the
yeast are unable to cause the lactose to ferment. However, when
lactase is present significant fermentation occurs. Lactase causes
lactose to split into glucose and galactose.

In conclusion, the yeast cells in the sealed flask underwent

fermentation to produce carbon dioxide and ethanol. In the
presence of oxygen, cells undergo aerobic cellular respiration. It uses
glucose in the presence of oxygen to produce CO2, H2O and ATP.
BIBLIOGRAPHY
• https://www.youtube.com/watch?v=FYClCHVT00M
• https://uwaterloo.ca/chem13-news-magazine/april-
2015/activities/fermentation-sugars-using-yeast-discovery-
experiment
• https://byjus.com/question-answer/name-one-commercial-use-
of-yeast/
• https://www.scientificamerican.com/article/single-celled-
science-yeasty-
beasties/#:~:text=Yeasts%20feed%20on%20sugars%20and,brea
d%20so%20soft%20and%20spongy.
• https://www.lidolearning.com/questions/s-bb-ncertexemplar8-
chp2-q11/name-one-commercial-use-of-yea/