MODERN MIDDLE EAST INTERNATIONAL

SCHOOL -RIYADH

CHEMISTRY PROJECT
ACADEMIC YEAR 2025-2026

SUBMITTED BY TEACHER IN-CHARGE

MOHAMED 12D Mrs Nadiya Anis
 ACKNOWLEDGEMENT
The work described in the project has been done with
assistance and guidance of my teachers.
I wish to thank Mrs. Nadiya Anis, my chemistry
teacher for her guidance and support for the completion
of the project.
I would like to extend my sincere thanks to Mrs.
Ayesha Touseef, our lab assistant for her constant
support.
I would like to express my gratitude and vote of thanks
to Mrs. Shabana Parveen, our Principal and Mrs.
Zainab Jaffri, our headmistress for their constant
encouragement and moral support.
Finally, I would like to thank my family and friends
for their source of inspiration.
 Certificate

This is to certify that the following healthcare

projects have been completed by Mohamed, a
student of 12d under the
guidance and supervision of the teacher as
per the requirement of the practical
examination 2025-26.

HEAD OF THE INSTITUTION TEACHER-IN-CHARGE

Mrs. Shabana Parveen Mrs. Nadiya Anis
Principal Chemistry teacher
Modern Middle East Modern Middle East
International School, Riyadh International School, Riyadh
INDEX
S. No. Content Page No.

1 Acknowledgement 2

2 Certificate 3

3 Objective 4

4 Introduction 5

5 History of Fermentation 7

Contributions to
6 9
Biochemistry & Uses of
Fermentation
7 Risks of Consuming 11
Fermented Food

8 Limitations of 12
Fermentation

9 Theory 13

10 Requirements 15

Experiment (Aim, Principle,

11 16
Preparation, Procedure)

12 Observation Table & 19

Calculations

13 Result 22

14 Conclusion 23

15 Graph 24

16 Precautions 25

17 Sources of Error 26

18 Bibliography 27
Objective:

The purpose of the experiment is to compare the rate of fermentation

of the given samples of wheat flour, gram flour, rice flour and
potatoes.
 Introduction:
Fermentation is a type of anaerobic metabolism which harnesses the redox potential
of the reactants to make adenosine triphosphate (ATP) and organic end products.
Organic molecules, such as glucose or other sugars, are catabolized and their
electrons are transferred to other organic molecules (cofactors, coenzymes, etc.).

Anaerobic glycolysis is a related term used to describe the occurrence of

fermentation in organisms (usually multicellular organisms such as animals) when
aerobic respiration cannot keep up with the ATP demand, due to insufficient
oxygen supply or anaerobic conditions. Fermentation typically is the conversion of
carbohydrates to alcohols and carbon
dioxide or organic acids using yeasts, bacteria, or a combination thereof, under
anaerobic conditions. A more restricted definition of fermentation is the chemical
conversion of sugars into ethanol. The science of fermentation is known as
zymology. Fermentation usually implies that the action of microorganisms is
desirable, and the process is used to produce alcoholic beverages such as wine, beer,
and cider. Fermentation is also employed in preservation techniques to create lactic
acid in sour foods such as sauerkraut, dry sausages, kimchi and yoghurt, or vinegar
for use in pickling foods.
History of fermentation
The art of fermentation is as old as the human civilisation on earth, as it was
traditionally developed by ancient societies for food preservation during harsh
seasons, for ritual feasts, and to enhance the sensory quality of food. Historical
records reveal that fermentation of several substrates, including milk and cereal, is
indigenous to many parts of the world. The earliest form of fermentation was
discovered by analysing the stone mortars from the Natufian burial sites of a semi-
sedentary foraging population, providing archaeological evidence for beer brewing
from cereals dating back 13,000 years In ancient Egypt, dairy products, fermented
bread, and beer were dietary staples . In China, chemical analysis of ancient pottery
jars indicate the existence of fermented products of rice, honey, and fruits as early as
the seventh millennium B.C. Wine production and fermentation have ancient
origins, with evidence found in Georgia (Caucasus) dating back 8,000 years. Jars
containing wine remains from 7,000 years ago were excavated in the Zagros
Mountains of Iran and are displayed at the University of Pennsylvania.

ZAGROS MOUNTAINS OF IRAN

Fermentation of beverages was practiced in various ancient civilizations: Babylon

(circa 5000 BC), ancient Egypt (circa 3150 BC), pre-Hispanic Mexico (circa 2000
BC), and Sudan (circa 1500 BC).
Contributions to biochemistry :
When studying the fermentation of sugar to alcohol by yeast, Louis Pasteur
concluded that the fermentation was catalyzed by a vital force, called "ferments,"
within the yeast cells. The "ferments" were thought to function only within living
organisms.
"Alcoholic fermentation is an act correlated with the life and organization of the yeast
cells, not with the death or putrefaction of the cells," he wrote. Nevertheless, it was
known that yeast extracts ferment sugar even in the absence of living yeast cells.
While studying this process in 1897, Eduard Buchner of Humboldt University of
Berlin, Germany, found that sugar was fermented even when there were no living
yeast cells in the mixture, by a yeast secretion that he termed zymase.

In 1907 he received the Nobel Prize in Chemistry for his research and discovery of
"cell-free fermentation."One year prior, in 1906, ethanol fermentation studies led to
the early discovery of NAD+.
Uses of fermentation:
1. Food Production
Bread making. Yeast ferments sugars in dough, producing CO₂ that makes bread
rise.
Dairy products. Yogurt, cheese, kefir, and sour cream are made through lactic
acid fermentation.
Pickling and preservation. Sauerkraut, kimchi, and pickles use fermentation to
preserve vegetables and improve flavor.

Soy products. Fermented soybeans produce miso, soy sauce, and tempeh.
2. Beverage Production
Alcoholic drinks. Beer, wine, and spirits are made by yeast fermentation of sugars.
Non-alcoholic drinks. Kombucha, ginger beer, and kvass are created through
controlled fermentation.
3. Industrial and Biofuel Applications
Ethanol production. Fermentation of sugars from crops like sugarcane or corn
produces bioethanol for fuel.
Acids production. Lactic acid, used in food and biodegradable plastics, citric acid,
a food additive, and acetic acid, which is vinegar.
Solvent production. Acetone and butanol are made through ABE fermentation.
4. Health and Medical Uses
Probiotics. Fermented foods like yogurt and kimchi contain beneficial bacteria
that support gut health.
Antibiotic production. Certain antibiotics, like penicillin, are produced using
fermentation technology.

penicillin

Vaccine manufacturing. Some vaccines are made using microbial

fermentation systems.
5. Scientific Research
Microbial studies. Fermentation is used in laboratories to study metabolism
and enzyme activity.
Biotechnology. It is used in genetic engineering to produce insulin, enzymes,
and other bio-products.
Risks of consuming fermented food
Food that is improperly fermented has a notable risk of exposing the eater to
botulism. Alaska has witnessed a steady increase of cases of botulism since 1985.
Despite its small population, it has more cases of botulism. This is caused by the
traditional Eskimo practice of allowing animal products such as whole fish, fish
heads, walrus, sea lion and whale flippers, beaver tails, seal oil, birds, etc., to
ferment for an extended period of time before being [Link] risk is
exacerbated when a plastic container is used for this purpose instead of the old-
fashioned method, grass-lined hole, as the botulinum bacteria thrive in the
anaerobic conditions created by the air-tight enclosure in plastic.

botulinum bacteria
How safe is it to consume fermented food?
Fermented foods generally have a very good safety record even in the developing world where the
foods are manufactured by people without training in microbiology or chemistry in unhygienic,
contaminated environments. They are consumed by hundreds of millions of people every day in
both the developed and the developing world. And they have an excellent safety record. What is
there about fermented foods that contribute to safety? While fermented foods are themselves
generally safe, it should be noted that fermented foods by themselves do not solve the problems of
contaminated drinking water, environments heavily contaminated with human waste, improper
personal hygiene in food handlers, flies carrying disease organisms, unfermented foods carrying
food poisoning or human pathogens and unfermented foods, even when cooked if handled or stored
improperly. Also improperly fermented foods can be [Link], application of the principles
that lead to the safety of fermented foods could lead to an improvement in the overall quality and
the nutritional value of the food supply, reduction of nutritional diseases and greater resistance to
intestinal and other diseases in infants.

Limitations of fermentation include:

Introduction of undesired microorganisms increases the risk of contamination, resulting in
lower yields or impure products
The need for specialized equipment and can increase the costs
Energy-intensive operations and the need for stringent conditions increase energy
consumption
The multifaceted nature of microorganisms can result in unexpected and undesirable end
products
The end product is often impure and has to undergo further purification treatment
Microbial cultures are likely to exhibit natural variations over time, potentially compromising
the quality and consistency of the end product
Theory
Wheat flour, gram flour, rice flour, potato juice and carrot juice contain starch as the major
constituent. Fermentation is the slow decomposition of complex organic compounds into simpler
compounds by the action of enzymes. Enzymes are biological molecules that catalyze (i.e, increase
the rates of) chemical reactions. Fruit and vegetable juices contain sugar such as sucrose, glucose
and fructose. The chemical equations below summarize the fermentation of sucrose, whose
chemical formula is C₁₂H₂₂O₁₁ . One mole of sucrose is converted into four moles of ethanol and
four moles of carbon dioxide:

Sucrose is hence first converted to glucose and fructose with the enzyme invertase, while enzyme
zymase converts glucose and fructose to ethyl alcohol.

Invertase
Invertase (systematic name: beta-fructofuranosidase) is an enzyme that catalyses the hydrolysis
(breakdown) of sucrose.
Related to invertases are sucrases. Invertases and sucrases hydrolyze sucrose to give the same
mixture of glucose and fructose. Invertases cleave the O-C (fructose) bond, whereas sucrases cleave
the O-C (glucose) bond. For industrual use, invertase is usually derived from yeast. It is also
synthesized by bees, who use it to make honey from nectar. Optimum temperature at which the
rate of reachon is al tis greatest is 600 C and an optimum ph of 4.5.
Zymase
Zymase is an enzyme complex ("mixture") which catalyzes the fermentation of sugar into ethanol
and carbon dioxide. They occur naturally in yeasts. Zymase activity varies among yeast strains.

Chemical test: Fehling's solution

To test for the presence reducing sugars to the juice, a small amount of Fehling' solution is added
and boiled in a water bath. During a water bath, the solution progresses in the colors of blue
(with no glucose present), green, yellow, orange, red, and then brick red or brown (with high
glucose present). A colour change would signify and the presence of glucose.
Sucrose (table sugar) contains two sugars (fructose and glucose) joined by their glycosidic bond
in such a way as to prevent the glucose isomerizing to aldehyde, or the fructose to alpha-
hydroxy-ketone form. Sucrose is thus a non-reducing sugar which does not react with Fehling's
solution. (Sucrose indirectly produces a positive result with Benedict's reagent if heated with
dilute hydrochloric acid prior to the test, although after this treatment it is no longer sucrose.)
The products of sucrose decomposition are glucose and fructose, both of which can be detected
by Fehling's as described [Link] comparing the time required for completion of fermentation
of equal amounts of different substances containing starch the rates of fermentation can be
compared.
Addition of yeast
In wine making, yeast is normally already present on grape skins. Fermentation can be done with
this endogenous "wild yeast," but this procedure gives unpredictable results, which depend upon the
exact types of yeast species present. For this reason, a pure yeast culture is usually added, this yeast
quickly dominates the fermentation. Baker's yeast is the common name for the strains of yeast
commonly used as a leavening agent in baking bread and bakery products, where it converts the
fermentable sugars present in the dough into carbon dioxide and ethanol. Baker's yeast is of the
species Saccharomyces cerevisiae, which is the same species commonly used in alcoholic
fermentation, and so is also called brewer's yeast.
Pasteur's salt
Pasteur's salt solution is prepared by dissolving ammonium tartarate, 10.0 g; potassium phosphate,
2.0 g; calcium phosphate, 0.2 g; and magnesium sulphate, 0.2 g dissolved in 860 ml of water.
The Pasteur's salts in solution act as a buffer to any acids the yeast may create. Since yeast only
converts sugar (most likely sucrose or glucose) to ethanol under anaerobic conditions, and it is
unreasonable to assume that there will be no oxygen present in the laboratory, some acetic acid is
created as a result. Pasteur salts act as buffers to prevent the proteins in yeast from denaturing due to
increased acidity caused by acetic acid production.
 Experiment:
AIM:
To compare the rate of fermentation of wheat flour, gram (besan) flour, rice flour, potato juice and
carrot juice and determine which sample shows the highest rate of fermentation under identical
conditions.

Requirements:
Conical flasks (6)
Active dry yeast
Gas syringes or inverted
Distilled or boiled cooled water
graduated cylinders and
Wheat flour
trough
Thermometer Gram flour (besan)

Stopwatch or clock Rice flour

Weighing balance Fresh potato juice
Measuring cylinders Fresh carrot juice
Hotplate or water bath
Amylase or malted barley powder
Glass rods and clamps

PRINCIPLE:
Yeast ferments available sugars into ethanol and carbon dioxide. The rate of
fermentation can be compared by measuring the volume of CO₂ produced over time
under identical conditions.
PREPARATION OF SAMPLES:
Prepare 10 percent w/v suspensions of flours and gelatinize by heating. If available, saccharify
flours with amylase or malted barley at about 37°C for 30–60 minutes. Prepare fresh potato and
carrot juice by blending and straining. Bring each sample to the same final volume and temperature
before adding yeast.

PROCEDURE:
Label six flasks: Glucose control, Wheat, Gram, Rice, Potato, Carrot.
Add 100 mL of each prepared sample into the respective flask.
Prepare yeast suspension by dissolving 1 g active dry yeast in 10 mL warm water at 37°C. Allow
to activate 5 minutes. Add 5 mL yeast suspension to each flask.
Attach gas syringes or arrange inverted graduated cylinders to collect CO₂. Ensure apparatus is
airtight.
Place flasks at constant temperature about 30–35°C. Start stopwatch immediately when yeast
is added.
Record CO₂ volume at 0, 5, 10, 15, 30, 45, 60 and 90 minutes.
Repeat each sample in triplicate in a real lab and use mean ± SD.

OBSERVATION TABLE ( CO₂ volume in mL):

CALCULATIONS
Method
Initial rate approximated as change in CO₂ from 0 to 15 minutes divided by 15 minutes. This gives
rate in mL CO₂ per min and is a simple way to compare initial fermentation activity.
Calculations for each sample using CO₂ at 15 minutes:
Glucose control
CO₂ at 15 min = 62 mL
Rate = 62 ÷ 15 = 4.133 mL per min
Carrot juice
CO₂ at 15 min = 48 mL
Rate = 48 ÷ 15 = 3.20 mL per min
Potato juice
CO₂ at 15 min = 45 mL
Rate = 45 ÷ 15 = 3.00 mL per min
Wheat flour
CO₂ at 15 min = 22 mL
Rate = 22 ÷ 15 = 1.467 mL per min
Gram flour
CO₂ at 15 min = 20 mL
Rate = 20 ÷ 15 = 1.333 mL per min
Rice flour
CO₂ at 15 min = 17 mL
Rate = 17 ÷ 15 = 1.133 mL per min
TABLE: INITIAL RATES (mL CO₂ per min )

Sample CO₂ at 15 min Rate (mL/min)

(mL)

Glucose control 62 4.13

Carrot juice 48 3.2

Potato juice 45 3

Wheat flour 22 1.47

Gram flour 20 1.33

Rice flour 17 1.13

RANK ORDER (highest to lowest initial rate)

1 Glucose control
2 Carrot juice
3 Potato juice
4 Wheat flour
5 Gram flour
6 Rice flour

RESULT:
Carrot juice produced the highest fermentation rate among the food samples. Potato
juice was close behind. The glucose control produced the highest rate overall,
confirming the yeast was active. Flours showed lower fermentation rates under the
conditions used.
 Graph:

CONCLUSION
Under identical conditions, carrot juice showed the highest rate of fermentation among the
tested food samples, followed by potato juice. Simple sugar in glucose gave the fastest
fermentation overall. Flour samples produced lower rates because their starch must be
converted to fermentable sugars before yeast can ferment them efficiently.
PRECAUTIONS:
Use the same amount and batch of yeast for all samples.
Maintain constant temperature for all samples.
Ensure the gas collection system is airtight.
Cool heated samples to about 37°C before adding yeast to avoid killing yeast.
Start timing promptly after yeast addition to keep experimental times consistent.

SOURCES OF ERROR:
Gas leakage results in underestimated CO₂ volumes.
Unequal yeast amounts or differing yeast activity.
Temperature variations between flasks.
Incomplete or unequal saccharification of flours.
Timing or reading errors when measuring volumes.
CO₂ dissolution in liquid reducing measured gas.
Contamination by other microorganisms.
BIBLIOGRAPHY:
1. Core Textbooks & General References:
Berg, J. M., Tymoczko, J. L., Gatto, G. J., & Stryer, L. (2022). Biochemistry (10th ed.). W.H. Freeman. (For
fundamental principles of glycolysis, fermentation pathways, enzyme kinetics, and NAD+ function).
Nelson, D. L., & Cox, M. M. (2021). Lehninger Principles of Biochemistry (8th ed.). W.H. Freeman. (For in-depth
details on metabolic pathways, including alcoholic and lactic acid fermentation).
Hutkins, R. W. (2018). Microbiology and Technology of Fermented Foods (2nd ed.). Wiley-Blackwell. (For
comprehensive information on food fermentation applications, safety, and microbiology).
2. Historical Context:
Barnett, J. A. (2003). A History of Research on Yeast 8: Taxonomy. Yeast, 20(11), 995–1015.
[Link] (For the history of yeast research and fermentation science).
McGovern, P. E. (2009). Uncorking the Past: The Quest for Wine, Beer, and Other Alcoholic Beverages. University
of California Press. (For archaeological evidence of ancient fermentation practices worldwide).
The Nobel Prize. (1907). Eduard Buchner - Nobel Lecture: Cell-Free Fermentation. [Link].
[Link] (Primary source for Buchner's groundbreaking
work).
3. Safety & Practical Applications:
National Center for Home Food Preservation. (2018). Fermenting: Safety and Guidelines. University of Georgia.
[Link] (Authoritative guide on the risks and safe practices for
food fermentation).
Marco, M. L., et al. (2017). Health benefits of fermented foods: microbiota and beyond. Current Opinion in
Biotechnology, 44, 94–102. [Link] (Scientific review on the health impacts of
fermented foods).
Centers for Disease Control and Prevention (CDC). (2021). Botulism: Prevention. U.S. Department of Health &
Human Services. [Link] (For information on botulism risks, particularly
related to anaerobic conditions).
4. Online Educational Resources:
Britannica, T. Editors of Encyclopaedia. (2023, October 24). Fermentation. Encyclopedia Britannica.
[Link]
LibreTexts™ Chemistry. (2022, February 14). 5.10: Fermentation.
[Link]
entation
ScienceDirect. (n.d.). Topics: Fermentation. Elsevier. [Link]
biological-sciences/fermentation (For accessing a wide range of scientific articles on the topic).
Thank Uou