Table of Contents

INTRODUCTION........................................................................................................................
1
OBJECTIVE..................................................................................................................... ...........
4
SCOPE AND LIMITATION............................................................................... ..........................
7
PRINCIPLE/THEORY...............................................................................................................
10
EXPERIMENT..........................................................................................................................
13
AIM:..........................................................................................................................................
13
REQUIREMENT:......................................................................................................................
13
PROCEDURE............................................................................................................................
16
OBSERVATION......................................................................................................................... 19
RESULT....................................................................................................................................
21
BIBLIOGRAPHY......................................................................................................................
22
 CERTIFICATE

This is to certify that ANUSHA PRASAD of Grade XII, AECS MAGNOLIA

MAARUTI PUBLIC SCHOOL, BANGALORE with
register number
has satisfactorily completed the project in Chemistry on
COMPARATIVE STUDY OF RATE OF FERMENTATION OF
FRUIT/VEGETABLE JUICES in partial fulfillment of the requirements of All
India Secondary School Certificate Examination (AISSCE) as prescribed by CBSE
in the year 2013-2014.

Signature of the Signature of the

Candidate Teacher In-Charge

Signature of the Signature of the

Principal External Examiner
 ACKNOWLEDGEMENT

I would like to thank my teachers, Mrs. Neelam and Mrs. Sowmya for
guiding me through this project and for their valuable inputs which provided
me with a constant nudge for improvement.

It is imperative to thank our Principal, Mrs. Seema Goel for providing

me the opportunity to work on this project.

It goes without saying that my classmates, especially Rochana Ramakrishna,

Pratusha Dinesh and Rahul for their help in due course of this project. My
parents have also played a part in helping me in this project. My thanks goes
out to them also.

This project and reading-up on the same has provided me with an in depth
understanding of the topic. It has nurtured my scientific temperament and
curiosity.

Signature of the

Candidate
 PRINCIPLE/THEORY

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  2 C6H12O6

Glucose + Fructose

+ Zymase  2 C2H5OH + 2CO2

C6H12O6
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.

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 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.

By comparing the time required for completion of fermentation of equal

amounts of different substances containing starch the rates of fermentation
can be compared.

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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 denatured.

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 EXPERIMENT

Aim:
To compare the rates of fermentation of some fruit/vegetable juices 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 reagent

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b. Apparatus Requirement

 Conical flasks  Test tubes  Beaker

 Bunsen burner, tripod stand and watch glass

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 PROCEDURE

1. 5.0 ml of apple juice was taken in a clean 250 ml conical flask and
diluted with 50 ml of distilled water.

2. 2.0 gram of Baker’s yeast and 5.0 ml of solution of Pasteur’s salts

were added to the above conical flask.
3. The contents of the flask were shaken well and the temperature of the
reaction mixture was maintained between 35-400C.

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4. After 10 minutes 5 drops of the reaction mixture were taken from the
flask and added to a test tube containing 2 ml of Fehling reagent. The
test tube was placed in a boiling water bath for about 2 minutes. The
colour of the solution or precipitate was then noted.

5. Step 4 was repeated after every 10 minutes until the reaction mixture
stopped giving any red colour or precipitate.

6. This time taken, i.e. time taken for the completion of fermentation was
noted.

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7. All the above steps were repeated by taking 5 ml each of grape juice,
black grape juice, sweet lime juice, orange juice and carrot juice.

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 Precautions:

 All apparatus should be clean and washed properly.

 The flask should not be rinsed with any of the
solution.

OBSERVATION

Volume of fruit juice taken = 5.0 ml

Volume of distilled water added = 50.0 ml
Weight of baker’s yeast added = 2.0 g
Volume of solution of Pasteur’s salts = 5.0
ml

Time Colour of reaction mixture on reaction with Fehling solution

( in
minutes ) Apple Sweet lime Carrot Orange Tomato
Juice Juice Juice Juice Juice

10 Red Red Red Red Red

20 Red Red Red Red Brownish
Red
30 Red Red No Change Red Brown
40 Red Red No Change Brown Dark Brown
50 Brownish Greenish No Change No Change No Change
Red Brown
60 Brown No Change No Change No Change No Change
70 No Change No Change No Change No Change No Change

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Graph

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 RESULT

The time taken for fermentation of carrot juice was well before the rest of
the juices, it’s recorded time being 30 minutes. This means that carrot juice
has the highest sucrose content from the various samples taken. After 50
minutes orange and tomato juices gave positive test for fermentation with
Fehling’s solution. For sweet lime juice time taken for fermentation was 60
minutes and for apple juice it was 70 minutes.

BIBLIOGRAPHY

Wikipedia - The free encyclopedia - (http://en.wikipedia.org)

Comprehensive Practical Chemistry

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