DAV PUBLIC SCHOOL

CHEMISTRY INVESTIGATORY
PROJECT
CLASS – XII

Topic – Comparative Study Of Rate Of

Fermentation Of Following Materials: Wheat
Flour, Gram Flour, Potato Juice, Carrot Juice
By- SHAISTA ZAREEN
ROLL NO- 25
 BONAFIDE CERTIFICATE

This is to certify that SHAISTA ZAREEN , a student of

classXII has successfully completed the research on
the topic of
“Comparative Study Of Rate Of Fermentation Of
Following Materials: Wheat Flour, Gram Flour,
Potato Juice, Carrot Juice” with parents, under the
guidance of
Mr NCH MURALIKRISHNA & Mr B SUDHAKER
during the year 2023-2024.

Signature of Examiner

Signature of Teacher

Signature of Principal
ACKNOWLEDGEMENT

This project report on

“Comparative Study Of Rate Of Fermentation
Of Following Materials: Wheat Flour, Gram
Flour, Potato Juice, Carrot Juice”
which also helped me in doing a lot of research
work and provided me the first-hand experience
of having such a joyful and educative task. I
consider myself fortunate enough to have the
valuable guidance of teachers
Mr NCH MURALIKRISHNA & Mr B SUDHAKER
for giving us chance to do my project work under
their surveillance. I sincerely stretch out thanks
to my venerable mates for their guidance and
encouragement in carrying out this project.
I would like to show my respect and gratitude to
Mr TVN SIVAKUMAR sir the honourable Principle
of this institution for setting up the opportunity
to mount this project.
 OBJECTIVE
In this project, time taken for fermentation of
various fruit/vegetable juices had to be
compared. Fermentation is one of the oldest
methods of processing food into a form that is
suitable for preservation
In fermentation technology, we stress in
understanding the various process in fermentor
and how various intrinsic factors influence the
fermentation process, Fermentation technology
being an industrial microbiology subject are
geared in producing maximum amount of high
economical fermentation products. The objective
of this project is to compare the rates of
fermentation of different fruit and vegetable
juices. The information gained. from this
experiment may be used by wineries to
determine which fruit juice ferments best. But it
is difficult to understand and control the
fermentation process as it involves various
components such as effect of substrates.
products inhibition, conditions and complex
microbial interactions. Fermentation is affected
by several factors including the temperature, salt
concentration, pH, oxygen availability and
nutrient availability. The rate of fermentation can
be controlled by manipulating any of these
factors.
Temperature
Different yeasts tolerate different temperatures.
For Saccharomyces cerevisiae, it is around 35-
40°C. A variation of just a few degrees from this
temperature alters the activity of the microbes
and affects the quality of the final product.
Nutrients i.e. Sugar content
All bacteria require a source of nutrients for
metabolism. The fermenters require
carbohydrates, in this case sugars glucose and
fructose. The energy requirements of microbes
are very high. Limiting the amount of substrate
available can reduce the rate of fermentation.
Effect of oxygen
If oxygen is present, some species of yeast will
oxidize pyruvate completely to carbon dioxide and
water. Thus, these species of yeast will produce
ethanol only in an anaerobic environment.
However, many yeasts such as the baker's yeast
Saccharomyces cerevisiae, or fission yeast
Schizosaccharomyces pombe, prefer fermentation
to respiration. These yeasts will produce ethanol
even under aerobic conditions.
Hence the rate of fermentation varies
The fermentation process is not only complex,
but always in a state of flux. Process, we are
therefore in a situation to always be adaptive and
reactive to these changes so that throughout the
fermentation process we are always staining the
conditions in a narrow window of optimal
fermentation conditions.
In order to help us do this we need to know
fermentation kinetics. When we talk about
fermentation kinetics we are talking about
fermentation modelu. Kinetics and modellings are
very useful to us as tools to make fermentation
predictions and enhancing our experimental
designs to be more focused to the specific
problems such as the rate limiting steps or
product inhibition
The study of fermentation kinetics helps us by
providing clear quantitative data for us to
understand the process and improve the process
accordingly. Peering into observation ports might
be good advertising gimmick for fermentation
technology but do not really help much in
understanding the process or even to control and
predict the fermentation outcome. Subjective
observations will rarely help in producing
optimum fermentation process and thus affect
profitability studies and making decisions.
Its numbers that count!
Thus, the importance of the study of
fermentation kinetics or models.
The first step in the study of fermentation kinetics
is to understand the various processes involved in
the whole process. Such questions such as inputs
and outputs, the metabolic pathways involved and
type of products or side products formed. The
various individual reactions involved and what
factors control the metabolite level. Then only
after all the relevant data are obtained do we start
formulating the models.
 INTRODUCTION
Fermentation is typically the conversion of
carbohydrates to alcohols and carbon dioxide or
organic acids using yeasts, bacteria, or a
combination thereof, under anaerobic conditions
(absence of oxygen) by the action of enzymes.
Enzymes are complex organic compounds,
generally proscias. They are highly specific with
regard to their substrates. Fermentation in simple
terms in the chemical conversion of sugars into
ethanol. Ethanol fermentation, also referred to as
alcoholic fermentation is the biological process in
which sugars such as ghase, fructose, and sucose
are converted into cellular energy and thereby
produce ethanol and carbon dioxide as metabolic
waste products. All ethanol contained in alcoholic
beverages is produced by means of fermentation
induced by yeast. Wine is produced by
fermentation of the natural sugars present in
grupes and other kinds of fruit. Ethanol
fermentation occurs in the production of alcoholic
beverages and ethanol fuel, and in the leavening of
bread dough. Fermentation is used in preservation
techniques and in production of foods such as
yogurt, cottage cheese (pancer), dhokla, idli,
chocolates, cheese ete. Fermentation has been
derived from the Latin word ferver, which means
to boil, at during fermentation, there is a lot of
frothing in the liquid due to evolution of carbon
dioxide. This gives it the appearance as if it is
boiling!

Yeasts are unicellular eukaryotic microorganisms

classified in the kingdom. Fungi, Yeast size can vary
greatly depending on the species, typically
measuring 3-4 m in diameter, although some
yeasts can reach over 40 mm. Most yeasts
reproduce asexually by mitosis, and many do so by
an asymmetric division process called budding.
Yausts do not form a single taxonomic or
phylogenetic grouping. The term yeast is often
taken as a synonym for Saccharomyces cerevisiae.

Natural fermentation precedes human history. The

curliest evidence of winemaking dates from eight
thousand years ago in Georgia, in the Caucas area.
Seven-thousand-year-old jars containing the
remains of wine have heen excavated in the Zagros
Misantains in Iran. There is strong evidence that
people were fermenting beverages in Babylon circa
3000 BC, ancient Egypt circa 3150 BC, pre-Hispanic
Mexico circa 2000 BC, and Sudan circa 1500 BC.
Ancient fermented food processes were
developed. long before man had any knowledge of
the existence of the microorganisma involved

When analying the fermentation of sigar to alcohol

by yeast, Louis Pasteur concluded that the
fermentation was catalyzed by a vital force, called
"ferments", within the youst cells. The "ferments"
were thought to function only within the yeast
cells. The ferments" were thought to function only
within living organism. Nevertheless, it was known
that yeast extracts (Yeast extract is the name given
to processed yeast products made by extracting
the cell contents (removing the cell walls) can
forment sugar even in the absence of living yeast
cells. While studying this process in 1897, Eduard
Buchner found that sugar was formented even
when there were no living yeast cells in the
mixture; by a yeast secretion that he termed
zymase. Le fermenting activity of yeast in due to
active catalyst of biochemical origin. In 1907 he
received the Nobel Prize in Chemistry for his
research and discovery of "cell-free fermentation."
Main uses of fermentation
The primary benefit of fermentation is the
conversion of sugars and other carbohydrates, eg,
converting juice into wine, grains into her
carbohydrates into carbon dioxide to leaven bread,
and sugars in vegetables into preservative organic
acids
Food fermentation has been said to serve five main
purposes:
• Enrichment of the diet through development of
a diversity of flavors, aromas, and textures in
food substrates.

• Preservation of substantial amounts of foods

through lactic acid, alcohol, acetic acid, and
alkaline fermentations

• Biological enrichment of food substrates with

proccia, essential amino acids, essential fatty
acids, and vitamins
• Elimination of antinutrients
• A decrease in cooking time and fuel
requirement
Risks of consuming fermented foods

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 than any other state in the
United States of America. 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 consumed. The 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.

Safety of Fermented Foods

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
contributes 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 unsafe. However,
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
One of the limitations of fermentation as a process
is its requirement for multiple reagents. Secondly,
in many cases the time taken is quite long and this
creates a need for catalyst. Without catalysts, the
reaction is extremely slow. The limitation of our
project is the slight error in the result and the
project is limited to the fermentation of the juices
with Baker's yeast and not under normal cond**-..
5 without adding Baker's yeast.

Owing to the different criterion on which the rate

of fermentation depends, if the experiment is not
carried out in the optimal temperature range, the
rates will turn out to be different than the actual
rates of the juices that have been taken. It is not
possible to get the exact theoretically estimated
value due to impurities in the reagents as well as
the compounds. Another point to be noted is that
the rates calculated from this experiment is just
one case and this can't actually access the rate of
fermentation of the fruit. An average needs to be
taken to access its actual value.
 PRINCIPAL/THEORY
Fermentation is the slow decomposition of
complex organic compounds into simpler
compounds by the action of enzymes Enzymes are
biological molecules that catalyze (ie, 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 C: HO. 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;
betafructofuranosidase) is an enzyme that
catalyzes 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 industrial 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 reaction is at its greatest is 60" C
and un 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's 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 above.
By 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, 20 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. The Pasteur salts act as
buffers to the acidity so that the proteins in the
yeast do not become denatured.
 EXPERIMENT
AIM: To compare the rates of fermentation of
wheat flour, gram flour, nice flour potato juice and
carrot juice and determine the substance which
has the highest rate of fermentation amongst the
various samples taken.
MATERIALS REQUIRED
• Conical flask
• Test tube
• Funnel
• Filter paper
• Water bath
• 1 % lodine solution
• Yeast
• Wheat flour
• Gram flour
• Rice flour
• Potato
• Aqueuos NaCl solution
PROCEDURE
• Take 5 gms of wheat flour in 100 ml conical
flask and add 30 ml of distilled water.
• Boil the contents of the flask for about 5
minutes.
• Filter the above contents after cooling, the
 filtrate obtained is wheat flour extract.
• To the wheat flour extract. taken in a conical
flask. Add 5 ml of 1% aq. NaCl solution.
• Keep this flask in a water bath maintained at a
temperature of 50-60 degree celsius. Add 2 ml
of malt extract.
• After 2 minutes take 2 drops of the reaction
mixture and add to diluted iodine solution.
• Repeat step 6 after every 2 minutes. When no
bluish colour is produced the fermentation is
complete. #Record the total time taken for
completion of fermentation.
• Repeat the experiment with gram flour extract,
rice flour extract, potato extract and record the
observations.
Observation

Time required for the fermentation-

Wheat flour-10 hours

Gram flour-12.5 hours
Pata juice 13 hours
Carrot juice-20 min

Conclusion

Carrot juice with the highest content of sucrose

among the given samples takes the least time to
get fermented.