INTRODUCTION :-

Fermentation is the slow decomposition of complex

organic compounds into simpler compounds by the action
of enzymes- Enzymes are complex organic compounds,
generally proteins- There are many examples of
fermentation processes which we come across in daily life;
souring of milk or curd, bread making, wine making and
brewing- Fermentation word has been
derived from Latin (Ferver which
means ‘to boil’). As during
fermentation there is lot of frothing of
the liquid due to the evolution of
carbon dioxide, it gives the
appearance as if it is boiling.

Louis Pasteur in 1860 demonstrated that

fermentation is a purely physiological process carried out
by living micro-organisms 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 cells. He proposed that
fermenting activity of yeast is due to active catalysts of
biochemical origin. These biochemical catalysts are called
enzymes. Enzymes are highly specific. A given enzyme
acts on a specific compound or a closely related group of
compounds.
Sugars like glucose and sucrose when fermented in the
presence of yeast Cells are converted to ethyl alcohol.
Sucrose is first converted to glucose and fructose with an
enzyme invertase. Enzyme zymase converts glucose and
fructose to ethanol.
 Sucrose
Invertase
C12H22O12+ H2O CH2OH
J
Sucrose
Glucose Fructose

C6H12O6
A Glycosidic
Glucose
Glucose or fructose bond
Fructose

During fermentation of starch, starch is first hydrolysed to

maltose by the action of enzyme diastase. The enzyme
diastase is obtained from germinated barley seeds.
Maltose is converted to glucose by enzyme maltase-
Glucose is converted to ethanol by another enzyme
zymase. Maltose

Diastase
2(C6H10O5)n + nH2O Diastase nC
►nC 1222H
12// O22
n
O11
Maltose
Starch
Maltase
C12H22O11 + H2O C6H12O6
Maltose Glucose

Zymase
C6H12O6 2C2H5OH + 2CO2
Glucose Ethyl Alcohol

Enzymes maltase and zymase are obtained from yeast.

 EXPERIMENT 1

To compare the rate of fermentation of the Apple Juice

and Orange juice

THEORY:-

The fruit and vegetable juices contain sugars such as

sucrose, glucose and fructose.These sugars on
fermentation in the
 presence of the enzymes invertase and zymase give ethanol via
the evolution of carbon dioxide.

Invertase C6H12O6 + C6H12O6

C12H22O12
Glucose Fructose
Sucrose

Ethanol
Zvmase
C6H12O6 + C6H12O6 • ►2C2H5OH + 2CO2
Glucose Fructose Ethanol

Glucose is a reducing sugar and gives red coloured

precipitates with Fehling solution, when warmed.
When the fermentation is complete, the reaction
mixture stops giving any red colour or precipitate OH O
with Fehling solution.

o OH
Fehling Reagent
REQUIREMENTS:-

1.Conica! flask (250 ml) ; 2. Test tube and Water bath

3.Apple juice ; 4.Orange juice ; 5.Fehling solution A and
Fehling solution B ; 6.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°- 4O°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 rereagent .

5. Place the test tube in boiling water bath for about 2

minutes and note the colour of the solution or precipitate.

6. Repeat the step 4 after every 10 minutes when the

reaction mixture stops giving any red colour or precipitate
with Fehling reagent, the completion of fermentation is
 indicated.

7. Note the time taken for completion of fermentation.

8. Repeat the same steps by taking 5-Oml of orange juice.

Pasteurs Salt Solution:- Pasteur salt solution is prepared by dissolving

ammonium tartrate lO.Og; potassium phosphate 2.0 g; calcium phosphate 0.2 g,
and magnesium sulphate 0.2 g, dissolved in 860 ml of water.
 OBSERVATIONS:-
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

Seri Time (in Colour of reaction mixture or reaction 'with

al min) fehling solution in case of
No. Apple Juice Orange Juice
1 10 Red Red

2 20 Red Red
3 30 Red Red
4
40 Red Brown
5 50 Brownish Red No Change
6 60 Brown No Change
7 70 No Change No Change

RESULT:-
The rate of fermentation of apple juice is more than the rate
of fermentation of orange juice.