TO COMPARE RATE OF FERMENTATION OF GIVEN

SAMPLE OF WHEAT FLOUR, GRAM FLOUR, RICE

FLOUR POTATO JUICE AND CARROT JUICE.
(A Project Report)

Submitted by
b

In partial fulfillment of the

Requirement for the AISSCE of Class XI

2019-2020

SUNFLAG SCHOOL
Zb b Bhandara road , warthi
 SRI VIJAY VIDYASHRAM SR.SEC. SCHOOL
VIJAY NAGAR, DHARMAPURI – 636701
(Affiliated to CBSE, New Delhi, Affiliation Code – 1930307)

BONAFIDE CERTIFICATE

BIOLOGY PROJECT

Certified project work done by ARUN.A of class XII in Sri Vijay Vidyashram

Sr. Sec. School 2019 – 2020.

Submitted for Central Board of Senior Secondary Education examination

held in Biology Laboratory in Sri Vijay Vidyashram Sr. Sec. School,

Dharmapuri.

Registration No :

Held on :

Center Code :

Internal Examiner Principal External Examiner

 ACKNOWLEDGEMENT

I wish to express my deep gratitude and sincere thanks to Principal,

for his encouragement and for all the facilities that he provided for
this project work. I sincerely appreciate this magnanimity by taking me into her
fold for which I shall remain indebted to his.

vv
I extend my hearty thanks to Vishal Borkar sir Chemistry teacher.
who guided me to the successful completion of this project. I take this opportunity
to express my deep sense of gratitude for his invaluable guidance, constant
encouragement, constructive comments, sympathetic attitude and immense
motivation, which has sustained my efforts at all stages of this project work.
I can’t forget to offer my sincere thanks to my classmates who helped me to carry
out this project work successfully and for their valuable advice and support, which
I received from them time to time.

bBhumika sahu
b

1
i I 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 foods.

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
2
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 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 Louis Pasteur
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
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 botulinum bacteria

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 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 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 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 O11. One mole of sucrose is converted into four moles of
ethanol and four moles of carbon dioxide:

C12H22O11 + H2O + Invertase gives 2 C6H12O6

(Glucose + Fructose)

C6H12O6 + Zymase gives 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

6
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 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 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.
 7

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, 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. The Pasteur salts
act as buffers to the acidity so that the proteins in the yeast do not become
8
denatured.

EXPERIMENT
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

b. Apparatus Requirement

Conical flasks
Test tubes

Beaker

stand

Bunsen burner, tripod

and watch glass

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PROCEDURE

1. Take 5 gm of wheat flour in 100 ml 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 5 ml of 1% aq. NaCl
solution.
5. Keep this flask in a water bath maintained at a temperature of 50-60 degree
Celsius and add 2 ml of malt extract.
6. After 2 minutes take 2 drops of the reaction mixture and add to diluted
iodine solution.
7. Repeat the above step after every 2 minutes. When no bluish colour is
produced the fermentation is complete.
8. Repeat the above steps for Gram flour too.
9. Take 5.0 ml of carrot juice and potato juice in two clean 250 ml conical flask
and dilute with 50 ml of distilled water separately.
10. Add 2.0 gram of Baker’s yeast and 5.0 ml of solution of Pasteur’s salts to the
above conical flasks.
11. After 10 minutes 5 drops of the reaction take the mixtures from the flask and
add to a test tube containing 2 ml 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 taken for completion of fermentation.

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 All apparatus should be clean and washed properly.
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 encyclopedia - ( http://en.wikipedia.org)
Comprehensive Practical Chemistry

Chemistry manual

www.icbse.com

12
 Index
S.No Topic Page number
1. Introduction 1
2. History 2
3. Contributions to biochemistry 3
4. Uses 4
5. Risks of consuming fermented foods 4
6. Safety of fermented foods 5
7. Limitations 5
8. Theory 6
9. Experiment 8
10. Bibliography 12