Vriksha Global School

Session: 2024-2025 Class: Grade 12

CHEMISTRY PROJECT
Roll No:

Topic: “To compare rate fermentation of give sample of wheat

flour gram, flour, rice flour and potato”

Submitted To: Submitted By:

C.R. Jayalakshmi Laksanaa.S.D. D
 INDEX
S.No Content Page

1 Certificate 3

2 Acknowledgement 4

3 Objective 5

4 Introduction 6

5 Materials Required 7

6 Procedure 8

7 Observations 9

8 Bibliography 10
 CERTIFICATE OF EXCELLENCE

This is to certify that Laksanaa.S.D. D student of grade 12 has

successfully completed the research on the below mentioned project
under the Guidance of Mrs. C R Jayalakshmi during the year of 2024 -
2025 in partial fulfilment of chemistry practical examination conducted by
Vriksha Global School.

Sign of the Sign of the

External Examiner Internal Examiner
ACKNOWLEDGEMENT

In the accomplishment of this project successfully, many people have best

owned upon their blessings and the heart pledge support, this time I am
utilizing to thank all the people who have been concerned with this
project.

Primarily I would like thank god for being able to complete this project
with success. Then I would like to thank me
Principal Ms.Yamini and my Chemistry teacher Mrs.Jayalakshmi whose
valuable guidance has been the one that helped me to patch this project
and make it full proof success, her suggestion and instruction has served
as the major contribution towards the completion of this project.

Then I would like to thank my parents who have helped me with their
valuable suggestions and guidance which has been very helpful in various
phases of the completion of the project.

Secondly, I would also like to thank my friends, and classmates who

helped me a lot in finalizing this project within the limited time frames.
INTRODUCTION
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 food.
HISTORY

Since fruits ferment naturally, fermentation precedes human

history. Since ancient times, however, humans have been
controlling the fermentation process. The earliest evidence of
winemaking dates from eight thousand years ago in Georgia, in
the Caucasus area. Seven thousand years ago jars containing
the remains of wine have been excavated in the Zagros
Mountains in Iran, which are now on display at the University of
Pennsylvania. There is strong evidence that people were
fermenting beverages in Babylon circa 5000 BC, ancient Egypt
circa 3150 BC, pre- Hispanic Mexico circa 2000 BC, and Sudan
circa 1500BC. There is also evidence of leavened bread in
ancient Egypt circa1500 BC and of milk fermentation in Babylon
circa 3000 BC. French chemist Louis Pasteur was the first known
zymologist, when in 1854 he connected yeast to fermentation. Pasteur
originally defined fermentation as "respiration without air".
Contributions to biochemistry
When studying the fermentation of sugar
to alcohol by yeast, Louis Pasteur
concluded that the fermentation was
catalysed 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 LOUIS PASTEUR
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.

YEAST
Uses
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 food through lactic

acid, alcohol, acetic acid and alkaline fermentations.

❖ Biological enrichment of food substrates with protein,

essential amino acids, essential fatty acids, and vitamins.

❖ Elimination of ant nutrients.

❖ A decrease in cooking times and fuel requirements.

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. 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.

BOTULINUM BACTERIA
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 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 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.
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 conditions i.e. 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.
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 C12 H22 011. One mole of sucrose is
converted into four moles of ethanol and four moles of carbon dioxide:

C12H22O11 + H2O + Invertase 2 C6H12O6

(Glucose + Fructose)

C6H12O6 + Zymase 2 C2H5OH + 2CO2

(Glucose + Fructose)

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
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 600 C and an optimum pH of 4.5.

Invertase
C12H22O11 + H2O Sucrose C6H12O6 + C6H12O6
(Sucrose) (Glucose) (Fructose)
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.
Zymase
C6H12O6 + C6H12O6 2C2H5OH +
2CO2
(Glucose) (Fructose Ethanol)
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 colours 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
glyosidic 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 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 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, 10.0g;
potassium phosphate, 2.0g; calcium phosphate, 0.2g; and magnesium sulphate,
0.2g dissolved in 860ml of water. The Pasteur’s salts in a 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 reasonable 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,
rice flour potato juice and carrot juice and determine the
substance which has the highest rate of fermentation amongst the
various samples taken.

Requirement
a. Chemical requirement
Pasteur’s salts

Yeast
Fehling’s solution

Test tubes
Beaker

Bunsen burner, tripod stand watch glass

PROCEDURE
1.Take 5gm of wheat flour in 100ml conical flask and add 30 ml
of distilled water.
2.Boil the contents of the flask for about 5 minutes.
3.Filter the above contents after cooling; the filtrate obtained is
wheat flour extract.
4.Take the wheat extract into a conical flask and add 5ml of 1%
aq. NaCl solution.
5.Keep this flask in a water bath maintained at a temperature
of 50-60 degree Celsius and add 2ml of malt extract.
6.After 2 minutes take 2 drops of the reaction mixture and add
to dilute iodine solution.
7.Repeat the above step after 2 minutes. When no bluish colour
is produced the fermentation is complete.
8.Repeat the above steps for gram flour too.

9.Take 5.0ml of carrot juice and potato juice in two clean 250ml
conical flask and dilute with 50ml of distilled water separately.
10.Add 2.0gram of Baker’s yeast and 5.0 ml of solution of
Pasteur’s salts to the above conical flasks.
25
11.After 10 minutes 5 drops of the reaction take the mixtures
from the flask and add to a test tube containing 2ml of Fehling
reagent. Place the test tubes in a boiling water bath for about 2
minutes. Note the colour of the solution or precipitate.
12.Record the total time take for completion of fermentation.

Precautions
 All apparatus should be clean and washed properly.
 The flask should not be rinsed with any of the
solution.

Observation
Time required for the fermentation__
Wheat flour--10 hours
Gram flour--12.5 hours
Potato 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

Bibliography:
Wikipedia – The free encyclopaedia - (http://en.wikipedia.org)

Comprehensive Practical Chemistry manual www.icbse.com