FERMENTATION

Fermentation: The decomposition of complex organic compounds into simpler substances

takes place in presence of living micro organisms. These changes are termed as Fermentation
Process
Merits of Fermentation Process:
1)
Cheaper raw materials may be used.
2)
Desired changes may brought about in one step, which would otherwise be produced
by a lengthy process.
3)
Its the only process for synthesizing complex compounds.
4)
It works under less drastic and economic conditions, compared to the chemical
process.
5)
Unwanted components may removed from the products.
Conditions favourable for Fermentation:
1)

2)
3)

4)

5)

Temperature: At very high temperatures the ferments are destroyed, while at very
low temperature the reaction is extremely slow. There is a certain optimum
temperature for every fermentation reaction, at which it proceeds most favourably.
Generally 25-30C is the most favourable temperature.
Presence of other substance: Certain nutritive solutions (generally inorganic salt
solutions) are added to act as food for the ferment cells.
Absence of Preservatives: Substances like boric acid and mercury salts, which retard
the fermentation reaction are called preservatives. Sine they destroy the ferments, so
should be absent.
Concentration: Ferments are rendered inactive by high concentration of the solution
to which these are added. The solutions used for fermentation should be sufficiently
diluted to favour the fermentation reaction.
Aeration: Mostly, the fermentation proceeds well only in the presence of air.

INDUSTRIAL APPLICATIONS OF FERMENTATION PROCESS:

Manufacture of Ethyl Alcohol by Fermentation:
Manufacture of Ethyl alcohol is of two types (i) from inexpensive starchy materials
(ii) from Molasses.
From Molasses: Molasses is syrup left after the separation of cane sugar or beet sugar crystal
from concentrated juice. Its a dark coloured, syrupy mass that still contains about 30%
uncrystalisable sucrose and about 32% over invert sugar (a mixture of glucose and fructose).
The different steps involved in the manufacture are:
(i)

Dilution: The molasses is diluted with water, until a concentration of 8-10% sugar is
obtained in solution. A small amount of dilute sulphyric acid is added so as to adjust
the pH of solution to about 4 to 4.5. The acidity is not only favourable for the growth

of yeast but it is also unfavourable for the growth of undesirable bacteria. Then to
this solution, a small quantity of ammonium sulphate is added which acts as a food
for the yeast cells.
Alcoholic Fermentation: The diluted solution is taken in big fermentation tanks and
some yeast is added (5% by volume of the liquid). The mixture is then allowed to
stand for a few days. The temperature is maintained at about 30C. Then the
fermentation starts and the enzymes invertase and zymase were produced by the
yeast. Then, invertase converts sucrose into glucose and fructose, which are then
converted into ethanol by another enzyme zymase.
Invertase
C12 H 22 O11 H 2 O

C 6 H 12 O6 C 6 H 12 O6

(ii)

Cane Sugar
Glucose
Fructose
Zymase
C 6 H 12 O6 2C 2 H 5 OH 2CO2
Glucose
Ethyl alcohol
The fermentation process is complete in three days and CO2 is liberated as the byproduct. The fermentated liquor called Wash (or) Wort.
(iii)

Fractional distillation: The fermented liquor (wash) possess the approximate

composition of 6 to 10% ethyl alcohol and 3 to 5% glycerol, higher alcohols (fuel oil)
and acetaldehyde. Then the wash is subjected to fractional distillation in a Still,
which consists of two fractionating columns, known as analyzer and rectifier, which
are provided with perforated plates. It works on counter-current principle, since
steam and alcohol travel in opposite direction through the still.
The wash is preheated by circulating the
coil round the rectifier and then introduced
into the analyzer. A current of steam is
passed from the bottom of the analyzer,
where by the alcohol present in the wash
rise up alongwith steam and enter the
rectifier. During the upward passage, steam
goes on condensing, while the vapours of
alcohols leaving from the top of the rectifier
and condensed and collected. This is
known as raw spirit, which contains about
90% alcohol (raw spirit).
The liquid
collected at the bottom of analyzer, is called

spent wash.
(iv)

Rectification: Ethyl alcohol, obtained above contains in addition to water various other
impurities which are removed by careful fractional distillation. The following fractions
are obtained first fraction containing low boiling liquids like acetaldehyde. Second
fraction containing 93-95% ethyl alcohol and is called rectified spirit. Last fraction
containing water and fuel oil.

(v)

Preparation of Absolute Alcohol: The rectified spirit contains 93-95% ethyl alcohol and
5-7% of water. Since it is a constant boiling mixture, so water cannot be removed simply
by distillation. Hence, some special methods are adopted for removing water from
rectified spirit so as to obtain 100% ethyl alcohol (i.e., absolute alcohol). To get absolute
alcohol, rectified spirit is treated with quicklime (CaO) and then distilled.
CaO H 2O Ca(OH ) 2

By products of alcoholic industry:

1)
Carbon dioxide (used for the preparation of dry ice)
2)
Acetaldehyde (Obtained in the first fraction)
3)
Fusel oil (Used for the preparation of amyl alcohol and amyl acetate)
4)
Spent wash (Used as a cattle food)
5)
Argol (Used for manufacturing of tartaric acid)

Preparation of Ethyl alcohol from Starch:

The manufacture of ethyl alcohol from starchy materials (potatoes, barley and maize) involves
the following steps:
1) Saccharification: The starch is first converted into maltose and this process is known as
saccharification. It completes in three steps.
a. Malting: Most of the barley is allowed to germinate in dark at 15C. Germinated
barley (called malt) is heated to 60C to stop further growth. It is then crushed and
extracted with water and filtered. The filtrate containing diastase enzyme, is
known as malt extract.
b. Mashing: It is the process of liberated starch from starchy materials. The starchy
materials is treated with superheated steam so as to break the cell walls and to
form a paste like mass called Mash.
c. Hydrolysis: Malt extract is added to mash at 50-60C, when the hydrolysis of the
starch into maltose takes places in about half an hours time.
Diastase
50 60 0 C

2(C 6 H 10 O5 ) nH 2 O
nC12 H 22 O11
Starch
Maltose
2) Alcoholic fermentation: The hydrolysed starch solution (containing maltose) is then
fermented by yeast for 2 to 4 days at 30C. The enzyme maltose (present in the yeast)
converts glucose to ethyl alcohol.
C12 H 22 O11 H 2 O Maltase
2C 6 H 12 O6
Maltose
Glucose
zymase
C 6 H 12 O6 2C 2 H 5 OH 2CO2
Glucose
Ethyl alcohol
3) Fractional distillation:
The fermented liquor (wash) possess the approximate
composition of 6 to 10% ethyl alcohol and 3 to 5% glycerol, higher alcohols (fuel oil)
and acetaldehyde. Then the wash is subjected to fractional distillation in a Still, which
consists of two fractionating columns, known as analyzer and rectifier, which are

provided with perforated plates. It works on counter-current principle, since steam and
alcohol travel in opposite direction through the still.
(Diagram is as above the
preparation of E.A. from Molasses)
The wash is preheated by circulating the coil round the rectifier and then introduced into
the analyzer. A current of steam is passed from the bottom of the analyzer, where by the
alcohol present in the wash rise up alongwith steam and enter the rectifier. During the
upward passage, steam goes on condensing, while the vapours of alcohols leaving from
the top of the rectifier and condensed and collected. This is known as raw spirit, which
contains about 90% alcohol (raw spirit). The liquid collected at the bottom of analyzer, is
called spent wash.
(4)

Rectification: Ethyl alcohol, obtained above contains in addition to water various other
impurities which are removed by careful fractional distillation. The following fractions
are obtained first fraction containing low boiling liquids like acetaldehyde. Second
fraction containing 93-95% ethyl alcohol and is called rectified spirit. Last fraction
containing water and fuel oil.

(5)

Preparation of Absolute Alcohol: The rectified spirit contains 93-95% ethyl alcohol and
5-7% of water. Since it is a constant boiling mixture, so water cannot be removed simply
by distillation. Hence, some special methods are adopted for removing water from
rectified spirit so as to obtain 100% ethyl alcohol (i.e., absolute alcohol). To get absolute
alcohol, rectified spirit is treated with quicklime (CaO) and then distilled.
CaO H 2O Ca(OH ) 2

Manufacture of Acetic acid by fermentation:

Acetic acid (CH3COOH) in the form vinegar (6-10% of acetic acid) is obtained by the
fermentation process, which is based upon the oxidation of fermented liquors (10-15% alcohol).
Fermentation is done by bacteril Mycoderma aceti in presence of air at 30-35C. The process is
termed as acetous fermentation
Its a slow process and takes about two to three
months for completion. A quick method has
been discovered, which is known as quick
vinegar process (German Process)
Procedure: Large wooden vats fitted with a
wood sharings impregnated with old vinegar (old
vinegar contains bacterium mycoderma aceti).
Dilute alcoholic solution containing the growth
nutrients (like phosphates and inorganic salts) is
allowed to trickle down through the sharings,
while an upward current of air enters through the
holes, near the bottom. The heat generated in the
oxidation of alcohol to acetic acid maintains the
temperature at 30-35C. The dilute acetic acid
solution obtained from first vat is then allowed to
trickle from the tops of a few other similar vats,

kept in series, till the concentration reaches about 8-10%. The process required 8-10 days for
compiletion.
Preparation of pure acetic acid (glacial acetic acid):
Acetic acid can be obtained from vinegar with the help of lime. The calcium acetate can be
crystalysed from the solution which yields pure acetic acid.
CaO 2CH 3COOH Ca (COOCH 3 ) 2 H 2 O
Distillati on
Ca (COOCH 3 ) 2 H 2 SO4
CaSO4 2CH 3COOH

Manufacturing of Acetone:
According to Weizmann process, starch (or) molasses is fermented with microorganism
clostridium acetobutylicum. Carbon dioxide and hydrogen are evolved during the reaction. The
products of fermentation reaction are butanol, acetone and ethanol, approximately in the ratio
6:3:1. These are separated and purified by distillation.
Manufacturing of Lactic Acid:
Lactic acid is prepared by the frmentation of sugar with lactic bacilli at 37-42C in the presence of
powdered chalk. Chalk helps to keep the acidity below 1% which is necessary because even 1%
free acid enders the bacilli inactive. Crystals of calcium lactate separate out and are removed.
Free acid is regenerated from the calcium salt by the addition of requisite quantity of dilute
sulphuric acid. The calcium sulphate is filtered off and the filtrate is distilled under reduced
pressure to get free lactic acid.
End-products obtained by Fermentation Process:
Types of end products of fermentation include:

Microbial cells (e.g. bacteria, yeast, fungal spores)

Microbial enzymes (e.g. milk clotting enzymes or rennets, recombinant fungal and
bacterial rennets for cheese manufacture)

Microbial metabolites (e.g. alcohols ethanol, butanol, 2, 3-butanediol, isopropanol;

chemicals lactate, propionate, proteins, vitamins, antibiotics; and fuels methane)

Recombinant products (e.g. hormones)