ABHINAV BHARTI HIGH SCHOOL

CHEMISTRY INVESTIGATORY PROJECT

Topic -STUDY THE RATES OF
FERMENTATION OF FRUITS OR VEGETABLE
JUICES
NAME-RUDRA JAISWAL
CLASS-12
SECTION-B
Roll- 12693608
 CERTIFICATE

This is to certify that Rudra Jaiswal of Class XII Science

has prepared the project on “Comparative Study Of
The Rate Of Fermentation Of Various Food Materials"

This project has been prepared under guidance of lab

instructor Biswanath sir and as per the
Norms provided by the CBSE.

………………………… …………………………
SUBJECT TEACHER EXAMINER
 ACKNOWLEDGEMENT
I would like to express my special thanks of gratitude to
my Chemistry teacher Mrs. Arpita Chakraborty for their
able guidance and support in completion of my project.

I would like to extend my gratitude to the Principal

Mrs. Sraboni Samanta and my Parents for providing me
with all the facility
that was required for this project.
 CONTENTS
->Introduction
->Theory
->Aim
->Apparatus
->Required
->Experiment
->Observation
->Result
->Precautions
->Bibliography
 FERMENTATION

What is Fermentation ???

Fermentation is a slow decomposition of complex
organic compound into simpler compounds by the
action of enzymes.Enzymes are generally protons
example of fermentation are souring of milk curd,
bread making wine making.
The word 'Fermentation' had been derived from latin
(Ferver means to 'boil'). As during fermentation there is
lot of frothing of liquid due to the evolution of carbon
dioxide, it gives the appearance as if it is boiling. Sugars
like glucose, sucrose when fermented in presence of
yeast cells are converted to ethyl.
alcohol. During fermentation of, starch is first
hydrolyzed to maltose by action of enzyme
diastase.Fermentation is carried out at a temperature
of 4-16°C (40-60°F). This is slow for most kinds of
fermentation, but is beneficial for cider as it leads to
slower fermentation with less loss of delicate. Apple
based juices with cranberry also make fine ciders and
many other fruit purees or flavourings can be used,
such as grapes, cherry, raspberry. The cider is ready to
drink after a three-month fermentation period, though
more after it is material in the vats for up to 2 to 3
years.Fermentation is a metabolic process that
converts sugar to acids, gases, or alcohol. It occurs in
yeast and bacteria, and also in oxygen-starved muscle
cells, as in the case of lactic acid fermentation.

Effect of oxygen
Fermentation does not require oxygen. If oxygen is
present,some species of yeast (e.g., Kluyveromyces
lactis or Kluyveromyces lipolytica) will oxidize pyruvate
completely to carbon dioxide and water. This process is
called cellular respiration. But these species of yeast
will produce ethanol only in an anaerobic environment
(not cellular respiration).
However, many yeasts such as the commonly used
baker's yeast Saccharomyces cerevisiae, or fission yeast
Schizosaccharomyces pombe, prefer fermentation to
respiration. These yeasts will produce ethanol even
under aerobic conditions, if they are provided with the
right kind of nutrition. During batch fermentation, the
rate of ethanol production per milligram of cell protein
is maximal for a brief period early in this process and
declines progressively as ethanol accumulates in the
surrounding broth. Studies demonstrate that the
removal of this accumulated ethanol does not
immediately restore fermentative activity, and they
provide evidence that the decline in metabolic rate is
due to physiological changes (including possible
ethanol damage) rather than to the presence of
ethanol. Several potential causes for the decline in
fermentative activity have been investigated.Viability
remained at or above 90%, internal pH remained near
neutrality, and the specific activities of the glycolytic
and alcohologenic enzymes (measured in vitro)
remained high throughout batch fermentation. None of
these factors appears to be causally related to the fall
in fermentative activity during batch fermentation.
Bread baking
The formation of carbon dioxide - a byproduct of
ethanol fermentation causes bread to rise. Ethanol
fermentation causes bread dough to rise. Yeast
organisms consume sugars in the dough and produce
ethanol and carbon dioxide as waste products. The
carbon dioxide forms bubbles in the dough, expanding
it into something of a foam.Nearly all the ethanol
evaporates from the dough when the bread is baked.

History of Fermentation
Since fruits ferment naturally, fermentation precedes
human history. Since ancient times, however, humans
have been controlling the fermentation process. The
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 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".

Louis Pastuer in 1860 demonstrated that fermentation

is apurely physiological process carried out by living
microorganism like yeast. This view was abandoned in
1897 when Buchner demonstrated that yeast extract
could bring about alcoholic fermentation in the
absence of any yeast cell. He proposed that fermenting
activity of yeast is due to active catalysts of biochemical
origin. These biochemicals are called enzymes.
Enzymes are highly specific compound or a closely
related group of compounds. Fermentation has been
utilized for many years in the preparation of beverages.
Materials from Egyption tombs demonstrated the
procedures used in making beer and leavened bread.
The history of fermentation, whereby sugar is
converted to ethanol by action of yeast, is also a history
of chemistry. Van Helmont coined the word iogaslt in
1610 to describe the bubble produced in fermentation.
Leeuwenhoek observed and described the cell of yeast
with his newly invented microscope in 1680. The fruit
and vegetable juices contain sugar such as sucrose,
glucose, and fructose. These sugars of fermentation in
presence of enzyme invert are and zymose give with
the evolution of CO2. Maltose is converted to glucose
by enzyme maltose, glucose to ethanol by another
enzyme zymose.

Importance of Fermentation
Enrichment of the diet through development of a
diversity
of flavours, aromas and textures in food substances.
Preservation of substantial amounts of food through
lactic
acid alcohol, acetic acids and alkaline fermentation.
Biological enrichment of food substances with
proteins,
essential amino acids, fatty acids and vitamins.
Elimination of anti-nutrients.
A decrease in cooking times and fuel requirements.

There are two types of respiration:

->Aerobic respiration - which needs oxygen
->Anaerobic respiration - which does not need oxygen

Fermentation or Anaerobic Respiration

Fruit juices contain various sugars like glucose, fructose
etc. When juices are treated with yeast and it converts
sugar into glucose and fructose. These
monosaccharides are further converted into ethyl
alcohol by another enzyme known as zymase.

The relative rates of fermentation can be established

with fillings solution A and B. Since glucose in an aldose
gives red precipitate with Fehling's solution. When all
the quantity of glucose is converted to ethanol, the
mixture will not give red colour.

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 0-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 60-0 C and an optimum pH
of 4.5.

Saccharomyces cerevisiae commonly called baker's

yeast is the chief strain used for the production and
purification of the enzyme. Invertase in nature exists in
different isoforms. In yeasts, it is present either as
extracellular Invertase or intracellular Invertase.
In plants, there are three isoforms each differing in
biochemical properties and subcellular locations.
Invertase in plants is essential not only for metabolism
but also help in osmoregulation, development and
defence system. In humans, the enzyme acts as an
immune booster, as an anti-oxidant, an antiseptic and
helpful for bone cancer or stomach cancer patients in
some cases. Zymase: Zymase is an enzyme complex
that catalyses the fermentation of sugar into ethanol
and carbon dioxide. It occurs naturally in yeasts.
Zymase activity varies among yeast strains. Zymase is
also the brand name of the drug pancrelipase. Zymase
is an enzyme complex ("mixture") which catalyses the
fermentation of sugar into ethanol and carbon dioxide.
They occur naturally in yeasts. Zymase activity varies
among yeast strains.

ADDITION OF YEAST
Yeasts can grow in the presence or absence of air.
Anaerobic growth, growth in the absence of oxygen, is
quite slow and inefficient. For instance, in bread dough,
yeast grow very little. Instead, the sugar that can
sustain either fermentation or growth is used mainly to
produce alcohol and carbon dioxide. Only a small
portion of the sugar is used for cell maintenance and
growth. In contrast, under aerobic conditions, in the
presence of a sufficient quantity of dissolved oxygen,
yeast grow by using most of the available sugar for
growth and producing only negligible quantities of
alcohol. This means that the baker who is interested in
the leavening action of carbon dioxide works under
conditions that minimize the presence of dissolved
oxygen. On the other hand, a yeast manufacturer that
wants to produce more yeast cell mass, works under
aerobic conditions by bubbling air through the solution
in which the yeast is grown. The problem posed to the
yeast manufacturer, however, is not as simple as just
adding air during the fermentation process. If the
concentration of sugar in the fermentation growth
media is greater than a very small amount, the yeast
will produce some alcohol even if the supply of oxygen
is adequate or even in abundance. This problem can be
solved by adding the sugar solution slowly to the yeast
throughout the fermentation process. The rate of
addition of the sugar solution must be such that the
yeast uses the sugar fast enough so that the sugar
concentration at any one time is practically zero. This
type of fermentation is referred to as a fed-batch
fermentation.
The role of yeast in winemaking is the most important
element that distinguishes wine from grape juice. In
the absence of oxygen, yeast converts the sugars of
wine grapes into alcohol and carbon dioxide through
the process of fermentation. The more sugars in the
grapes, the higher the potential alcohol level of the
wine if the yeast are allowed to carry out fermentation
to dryness. Sometimes winemakers will stop
fermentation early in order to leave some residual
sugars and sweetness in the wine such as with dessert
wines. This can be achieved by dropping fermentation
temperatures to the point where the yeast are inactive,
sterile filtering the wine to remove the yeast or
fortification with brandy to kill off the yeast cells. If
fermentation is unintentionally stopped, such as when
the yeasts become exhausted of available nutrients,
and the wine has not yet reached dryness this is
considered a stuck fermentation.

ALCOHOLIC FERMENTATION:
It is a biological process which converts sugars such as
glucose, fructose, and sucrose into cellular energy,
producing ethanol and carbon dioxide as a side-effect.
Because yeasts perform this conversion in the absence
of oxygen, alcoholic fermentation is considered an
anaerobic process. Ethanol fermentation has many
uses, including the production of alcoholic beverages,
the production of ethanol fuel, and bread baking.

1. ACID FERMENTATION:
Lactic acid fermentation is a biological process by which
glucose and other six-carbon sugars (also, disaccharides
of six-carbon sugars, e.g., sucrose or lactose) are
converted into cellular energy and the metabolite
lactate. It is an anaerobic fermentation reaction that
occurs in some bacteria and animal cells, such as
muscle cells. Sometimes even when oxygen is
present and aerobic metabolism is happening in the
mitochondria, if pyruvate is building up faster than it
can be metabolized, the fermentation will happen
anyway. in homolactic fermentation, one molecule of
glucose is ultimately converted to two molecules of
lactic acid. Heterolactic fermentation, in contrast, yields
carbon dioxide and ethanol in addition to lactic acid, in
a process called the phosphoketolase pathway.
AIM:- To compare the rates of fermentation of the
following fruit or vegetable juices (i) Apple juice (ii)
Orange juice (iii) Carrot juice

REQUIREMENTS
Conical flasks (250 mL), test tubes and water bath,
Apple juice, Orange juice, Carrot juice, Fehling solution
A, Fehling solution B, solution of Pasteur salts and
distilled water.
PROCEDURE
1. Take 5.0 mL of apple juice in a clean 250 mL conical
flask and dilute it with 50 mL of distilled water.
2. Add 2.0 g of Baker’s yeast and 5.0 mL of solution of
Pasteur’s salts to the above conical flask.
3. Shake well the contents of the flask and maintain the
temperature of the reaction mixture between 35°–
40°C.
4. After 10 minutes take 5 drops of the reaction
mixture from the flask and add to a test tube
containing 2 mL of Fehling reagent. Place the test tube
in boiling water bath for about 2 minutes and note the
colour of the solution or precipitate
[Pasteur’s Salt Solution Pasteur salt solution is prepared
by dissolving ammonium tartrate, 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.]
Observation
All fruit juices do not undergo fermentation at the
same rate.The increasing order of the rate of
fermentation. carrot juice <orange juice < apple Juice.

Result
The rate of fermentation is carrot juice <orange juice <
apple Juice.
 BIBLOGRAPHY

Practical Chemistry Comprehensive - XIl

Class XII Chemistry Text Book
Internet
www.google.com
http://fermentationtechnology.blogspot.com
https://en.wikipedia.org/wiki/Fermentation
articles.mercola.com/fermented-foods.aspx
draxe.com/fermented-foods
www.morethanorganic.com/fermentation
AIM
To compare the rates of fermentation of the given
samples of wheat flour, gram flour, rice and potatoes.
REQUIREMENTS Conical flasks, test tubes, funnel, filter
paper and water bath. Wheat flour, gram flour, rice
flour, potatoes, 1% iodine solution.
PROCEDURE
1. Take 5.0 g of wheat flour in a 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. To the wheat flour extract taken in a conical flask,
add 5 mL of 1% aqueous NaCl solution.
5. Keep this flask in a water bath maintained at a
temperature of 50–60°C. Add 2 mL of malt extract.
6. After 2 minutes take 2 drops of the reaction mixture
and add to diluted iodine solution. Note the colour
produced.
7. 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.
8. Repeat the experiment with gram flour extract, rice
flour extract, potato extract and record the
observations.
OBSERVATION
Time required for the fermentation—-
# Wheat flour —10hours
# Gram flour—12.5hours
# Rice flour — 15 hours
#Potato—13 hours

CONCLUSION
Rice flour takes maximum time while wheat flour takes
the least.