The Fermentation Industry

Introduction

The term fermentation (Latin fermentare to boil) formerly stood for decomposition of food stuff usually
accompanied by evolution of gas. The fermentation of sugars to alcohols and carbon dioxide by yeast is
one of the oldest example. The term fermentation is now applied to changes brought about by micro
organisms. Evolution of gas is not an essential criterion. In our daily life we see many complex chemical
reactions, which are brought about by the agency of living organisms. Examples arc, souring and curdling
of milk, putrefaction of meat, production of indigo dye fruin the compound indicate in the food, curing of
tobacco, the development of benzaldehyde or oil of bitter almonds from the amygda contained in the
almond seed, conversion of fruit juices into wines etc. All these processes are called fermentation
processes, in which complex organic material is broken down into smaller substances and decomposition
is brought about by the action of living organisms, which secrete the enzyme catalyst suitable to the
process.

Although the fermentation of fruits to alcohol and making of beverages out of fruits and grams have been
established for centuries, it is only dung the past generation that the wider application of fermentation
has been recognized. Now man is directing the life process of micro-organisms the yeasts, bacteria and
moulds, to the production of large number of chemicals, such as alcohol, acetone, acetic acid, lactic acid,
citric acid and many antibiotics which are of great synthetic as well as industrial importance

Although the fermentation of fruits to alcohol was known to primitive humans, and although the making
of various beverages out of fruits and grains has been well established for centuries, only during the past
century has wider application of this procedure been recognized. Now scientists are directing the life
processes of yeasts, bacteria, and molds to produce chemicals. The modern definition of industrial
fermentation is any microbial process controlled by humans that produces useful products.

The foundation of the scientific understanding of fermentation, indeed of the action of all
microorganisms, hence of their economic control, rests firmly upon the genius of one man, Louis Pasteur.'
He showed that fermentation is directly caused by the life processes of minute organisms. By
understanding how these microorganisms function and by recognizing that varieties of yeasts, for
example, act differently and that the environment fundamentally affects even a given strain, these
processes of fermentation can be controlled in an exact scientific manner. Microorganisms, which include
bacteria, yeasts, and molds, feed upon organic materials. It is this feeding that interests the manufacturer,
for if they are supplied with the necessary energy foods, together with other needed nutrients, these
micro vegetative organisms will not only grow and multiply but will change the food into other chemical
substances.
Yeasts and bacteria are unicellular and of very small dimensions. Yeasts are irregularly oval and around
0.004 to 0.010 mm in diameter. Bacteria are smaller, mostly less than 0.007 mm in the longer dimension,
and more diverse in shape. Many of them, bacilli, are rod-shaped. Yeasts multiply by budding, and bacteria
by binary fission. Molds are multicellular filaments and increase by vegetative growth of the filament.
Sporulation provides for the next cycle, as it does also with many bacteria. The vegetative reproduction
cycle of these bacteria and of yeast is short-measured in minutes. Because of this, they multiply
exceedingly fast. The production of lactic acid in 1880 was the beginning of industrial fermentation to
produce a useful product other than alcohol. During World War I, Chaim Weizmann developed a
fermentation process to convert corn to acetone and n-butanol. Acetone was necessary for the
manufacture of cordite for use in ammunition and without this fermentation process Brit- ain would have
had a serious shortage of ammunition. Citric acid and gluconic acid were successfully produced between
1920 and 1940, but the young petrochemical industry then made most fermentation processes
uneconomical. During World War II, the discovery of antibiotics, such as penicillin, set the stage for the
great technological advances in controlling microbiological processes that are commonly used today

USES AND ECONOMICS

Many fermentation processes are in direct competition with strictly chemical syntheses. Alcohol, acetone,
butyl alcohol, and acetic acid produced by fermentation have largely been superseded by their synthetic
counterparts. Almost all the major antibiotics are obtained from fermentation processes. Dextran is
another fermentation product. The microbiological production of vitamins has also become economically
important. In Chap. 40 antibiotics, hormones, and vitamins are presented together with several
flowcharts. The enzyme glucose isomer is used to produce high fructose syrup from corn, and proteases
produced from mold enzymes are used to coagulate protein in milk to produce cheese.

Fermentation under controlled conditions involves chemical conversions. Some of the more important
processes are: oxidation, e.g, alcohol to acetic acid, sucrose to citric acid

Condition favourable for the fermentation process

The most important factor that influence the process o f fermentation are-

1) Temperature: The enzymes are destroyed at very high temperature. Most of them become
inactive above 80C. The process is, however very slow below 20C. So the temperature should be
neither too low nor too high. Generally, an optimum temperature of 30-50C is most favourable.
2) Aeration: Fermentation process usually pocked well only in the presence of air.
3) Concentration: High concentration of a solution renders an enzyme inactive. Thus the solution
used in fermentation should be sufficient dilute to favour the process.
4) Presence of other substances: Certain inorganic salt solution act as food for the ferment cell.
5) Absence of preservatives: Preservatives are those substances which destroy the ferment and
retard the fermentation reaction. Hence these substances should be absent.
Characteristics of enzymes in fermentation industry
Enzymes play a crucial role in the fermentation industry. Here are some of their key characteristics:

1) Biocatalysts: Enzymes act as biological catalysts, accelerating the rate of chemical reactions in
fermentation processes without being consumed themselves.
2) Specificity: Each enzyme is highly specific to its substrate, meaning it will only catalyze a particular
reaction or a specific group of reactions.
3) Temperature and pH sensitivity: Enzymes have optimal temperature and pH ranges at which they
function most efficiently. Deviations from these ranges can impact their activity.
4) Substrate concentration: Enzymes exhibit saturation kinetics, where their activity increases with
increasing substrate concentration up to a certain point, beyond which further increases do not
impact the reaction rate.
5) Enzyme concentration: The rate of reaction is directly proportional to the enzyme concentration
until all substrate molecules are converted.--Denaturation: High temperatures or extreme pH
levels can cause enzymes to denature, losing their structure and activity.
6) Immobilization: In industrial applications, enzymes can be immobilized onto surfaces or carriers
to improve their stability and reusability.--Enzyme inhibition: Various factors, such as product
accumulation or the presence of specific molecules, can inhibit enzyme activity.
7) Fermentation optimization: Enzymes are used in fermentation processes to break down complex
substrates into simpler ones, converting sugars into alcohol, acids, or other products.
8) Industrial scale production: Enzymes used in the fermentation industry are typically produced
through microbial or recombinant DNA technologies, allowing for large-scale production.

Overall, enzymes offer remarkable benefits in the fermentation industry by providing specificity,
efficiency, and control in various bioprocesses.

Types of Fermentation Processes

Batch Fermentation

In batch fermentation, all the components are mixed at once then the
reaction undergoes without any further intake from outside. During the
whole process, no extra nutrients are added. It is a closed system
because all the components are added at once and no other
components are added in between the process of fermentation.
There are three phases in the batch fermentation process - lag phase,
exponential phase, and stationary phase.
 • In the lag phase, microbes adapt to the environment of the culture
and

• In the exponential phase, the microbial cells grow rapidly and

consume most of the nutrients and the last phase is

• The stationary phase is when the growth of microbes stops due to

the consumption of all nutrients. It is the simplest type of all
industrial fermentation.

Fed-Batch Fermentation

• It is a modification of batch fermentation.

• In this nutrition is added aseptically and the amount of liquid

culture in the bioreactor increases as the culture is added
systematically.

• It is a type of semi-open system.

• It yields a better result than batch fermentation.

• After consumption of early substrate continuous and constant

nutrition is added.

Continuous Fermentation

• It is a type of fermentation in which constant addition and flow of

solution occur.

• Microorganisms and sterile nutrients are added continuously and

the nutrient solutions and microbes are transformed
simultaneously.

• It is a type of open fermentation system in which comments can be

added and removed in between the process.
 • There are many methods of continuous fermentation.

Microorganisms Used in Fermentation

• Yeast: Yeast is a single-celled fungus used in various fermentation processes. It plays a crucial role
in alcohol fermentation, converting sugars into ethanol and carbon dioxide. This process is widely
used in brewing beer, wine-making, and producing spirits like whiskey and rum. Yeast is also
essential for bread-making, where it produces carbon dioxide gas, causing the dough to rise.
• Lactic Acid Bacteria (LAB): LAB are a group of bacteria that convert sugars into lactic acid during
fermentation. This process is used in the production of various dairy products, including yogurt,
cheese, and buttermilk. The fermentation of lactose by LAB enhances the flavor, texture, and shelf
life of these products.
• Acetic Acid Bacteria: Acetic acid bacteria are responsible for the fermentation of alcohol into
acetic acid, which is the primary component of vinegar. These bacteria oxidize ethanol to acetic
acid and are commonly used in the production of different types of vinegar, such as apple cider
vinegar and balsamic vinegar.
• Mold and Bacteria for Soy Sauce and Tempeh: The fermentation of soybeans to produce soy sauce
and tempeh involves specific molds (e.g., Aspergillus oryzae for soy sauce) and bacteria (e.g.,
Rhizopus oligosporus for tempeh). In soy sauce production, the mold breaks down the proteins
and carbohydrates in the soybeans to form a flavorful liquid. For tempeh, the mold binds the
soybeans into a dense, cake-like form.

These microorganisms are essential in fermentation processes, as they convert sugars and other organic
compounds into various desirable products, adding flavor, preserving food, and creating unique textures.
Their controlled use allows for the production of a wide range of fermented foods and beverages with
distinct characteristics and cultural significance.

Fermentation substrate
Fermentation substrates are the organic compounds or materials that microorganisms use as a food
source during the fermentation process. The type of substrate used can vary depending on the
microorganism and the desired end product. Here are some common fermentation substrates:

Sugars: Various sugars are commonly used as fermentation substrates. Glucose, sucrose, fructose, and
maltose are frequently utilized by microorganisms like yeast to produce alcohol, carbon dioxide, and other
byproducts in processes such as brewing and wine-making.

Lactose: Lactose, the primary sugar in milk, is fermented by lactic acid bacteria (LAB) to produce lactic
acid. This is the basis for fermenting dairy products like yogurt, cheese, and buttermilk.
Ethanol: Ethanol, an alcohol, can serve as a substrate for acetic acid bacteria to produce acetic acid during
vinegar fermentation.

Soybeans: Soybeans are used as a substrate for fermentation processes such as soy sauce and tempeh
production. Specific molds and bacteria break down the proteins and carbohydrates in soybeans to create
the final products.

Starch: Starchy materials like grains (e.g., barley, rice, corn) or tubers (e.g., cassava, potatoes) can be
converted into sugars, which serve as substrates for various fermentation processes.

Cellulose: Some microorganisms can ferment cellulose, a complex carbohydrate found in plant cell walls.
This process is used in certain biogas production and composting applications.

Fruits and Vegetables: Fermentation of fruits and vegetables can yield a variety of products, such as
fermented pickles, sauerkraut, kimchi, and fruit preserves.

Molasses: Molasses, a byproduct of sugar refining, contains sugars and other nutrients suitable for
fermentation. It is often used in industrial fermentation processes.

These substrates provide the necessary nutrients for microorganisms to thrive and undergo metabolic
processes, converting the organic compounds into various products through fermentation. The choice of
substrate and microorganism determines the final outcome, flavor, and characteristics of the fermented
product

SOME FERMENTATION PROCESSES

Digestion of food is a very complicated process depending on the consecutive influences of a whole series
of enzymes. The enzymes, pepsin and ptyalin (origin: saliva) in gastric juice work together to increase the
rate of breaking down of large molecules of protein and starch into small ones. The lare are easily utilised
by the cells of the body. Enzymes are thus all important in the control of biological processes.

Alcohol is produced by the action of zymase enzyme on certain sugars, the almost important of which is
glucose, which is found in sweet fruits and honey along with fructose. Zymase is secreted by the m
organism known as yeast. When yeast is added to a solution of glucose, the latter decomposes into alcohol
and carbon dioxide. The decomposition is brought about through the agency of zymase.

C6H12O6 + (Zymase) → 2C2H5OH + CO2

It should be noted that all sugar cannot be fermented by zymase. Cane sugar or beat sugar and wheat
sugar or maltose is not fermented by zymase.

These sugars are also fermented by adding yeast into the solution of these sugars Fermentation in takes
place due to the fact that the yeast secrets not only the zymase, but also the enzymes invertase and
maltase. Invertase brings about the conversion of cane sugar or beat sugar or sucrose into glucose and
fructose, by a process known as hydrolysis.

C12H22O11(sucrose) +H2O (Invertase) → C6H12O6 (glucose) + C6H12O6(fructose)

n.C12H22O11(maltose) +H2O (maltase) → 2C6H12O6 (glucose)

Fermentable sugars can also be obtained from starch. The latter is derived mainly from rice, wheat and
maize

The enzyme, diastase is contained in malt. The latter is germinated barley or wheat. The malt is produced
by allowing the steeped (in water) barley to germinate for 7-10 days at temperatures 10'C -15°C and then
heated for stopping the germination. The barley grains are first cleaned and graded and then steeped for
some days in water at a temperature of 10-15°C. The steeped barley is then spread out or "couched" to a
depth of two or three feets on the floor of the malt house. As the germination proceeds, he moist grains
soon become to sprout and to become hot With the advance of germination, heat is evolved and the
temperature tends to rise. Hence, the couch or pile is broken down and the germinating barley spread
out in a thin layer, only a few inches in depth. The depth of the pile is carefully controlled. so that
temperature may not become too low or too high. For proper germination, the pile or couch is turned
over from time to time so as to allow access of air to the grains and sprinkled with water from time to
time when necessary. During the germination of the barley, diastase and other enzymes are produced
and when growth has proceeded sufficiently fare, the process of germination is stopped by allowing the
rootlets to wither, (i.e. rootless are allowed to fade away by withdrawing moisture from the germinating
grain) and then drying the malt in a kiln. After withering, which requires about 24 hours, the green malt
is hoisted on the floor or a kiln. The green malt is piled on the floor 12-18 inches deep and frequently
turned over for drying-and curing. Drying, which requires about 2 days, is first carried out at about 27°C
and then temperature is allowed to increase gradually to 38°C. After drying, the temperature is increased
to about 70'C and then allowed to rise to 95°C. This process is called curing. Killed malt is obtained in this
manner. It is cooled and stored in bins with less than 3% moisture.

The malt, which contains a considerable amount of starch and a small amount of sugar, together with the
enzyme diastase, is crushed and mixed with hot water and raw grain or potato starch is then added. The
temperature is maintained between 40-60°C. As a result of fermentation in presence of diastase, starch
is converted into maltose or malt sugar. This process is called mashing. After the completion of mashing,
the sweet liquid or wort is then conveyed to the fermenting vats and yeast is added. The enzyme maltase
present in the yeast converts maltose into glucose. The latter is converted into alcohol and carbon dioxide
by the enzyme zymase also present in yeast. Small amounts of higher alcohols, succinic acid, glycerine c.
are also produced.
When fermentation is carried out at about 15°C, a turbulent effervescence is produced due to the
liberation of CO2 gas. The liberation of CO2 is so rapid that the yeast cells are carried to the surface of the
liquid and as a result, a thick froath is formed. This process is known as top fermentation.

When fermentation is allowed to proceed at a low temperature, say at about 6°C.the evolution of carbon
dioxide is too slow to carry the yeast cells to the surface of the liquid. The yeast cells, therefore, main at
the bottom of the vat. This process is called bottom fermentation.

MANUFACTURE OF POWER ALCOHOL

The various important raw materials for the manufacture of power alcohol or ethyl alcohol are accharine
materials (such as molasses, sugar beets, sugar cane etc), starchy materials such as starch, potatoes,
cereal grains cc), cellulosic materials (such as sulphite liquor from paper mills) and hydrocarbon gases.
ETHYL ALCOHOL FROM MOLASSES
Molasses is a dark coloured viscous liquid left after the crystallization of cane sugar from cane juice. It
contains 50-55% total sugars (cane sugar. glucose and fructose) of which 35-40% is cane sugar sucrose.
Black strap molasses is concentrated cane juice, from which no cane sugar has been extracted

Molasses is converted into ethyl alcohol by means of yeast, which contains enzymes invertase and zymase
responsible for fermentation

C12H22O11(sucrose) +H2O (Invertase) → C6H12O6 (glucose) + C6H12O6(fructose)

C6H12O6 + (Zymase) → 2C2H5OH + 2CO2

25 gallons of molasses containing about 52% of sugar content have been found to produce about one
gallon of 95% ethyl alcohol.

PREPARATION OF WASH
Molasses, because of strong concentration of sugar, does not support direct yeast fermentation. The
molasses is, therefore, diluted to a concentration of 10-15 percent sugars. If concentration is high, more
alcohol may be produced and may kill the activity of the bacteria or yeast. If the concentration is low,
large quantity of wort or fermented liquor is to be diluted to get the same quantity of alcohol,

A small amount of dilute sulphuric acid is also added to check the growth of undesirable bacteria, which
would otherwise form lactic acid and acetic acid. The pH of the solution is also brought down to 4-5-5.0
with sulphuric acid. Some nutrients for yeast, such as ammonium phosphate are also added, if sufficient
yeast fold is not already present. The resulting solution is called mash. It is pumped to a large steel
fermenter, closed in moderh plant, to collect the CO, evolved and to afford easier conditions for cleaning
and sterilizing. A charge of selected yeast (about 5% of the total volume) is added to the fermentor and
the temperature is maintained at about 30°C. The reaction is exothermic and cooling coils are necessary
to maintain the temperature. Fermentation seis in and enzymes present in the yeast bring about the con
version of sucrose and other sugars into ethyl alcohol. The enzyme invertase brings about the hydrolysis
of sucrose into glucose and fructose. The yeast also produces another enzyme, zymase, which changes
the mono saccharides glucose and fructose into ethyl alcohol and carbon dioxide. In about 48 hours, 8-10
percent alcohol is produced. The carbon dioxide liberated in the process is scrubbed with water and then
sold as dry ice or otherwise disposed off. The fermentation is arrested by passing air through the liquid
The yeast is filtered off and the filtrate thus obtained is called wash or wort and contains about 6-10
percent alcohol.
The various stages of fermentation are:

In first few hours yeast cells multiply. The optimum temperature for yeast propagation is 86F-88°F. Rapid
fermentation then takes place and oxygen is given off rapidly. This process is called boiling, which takes
place at 90-95°F. The process of boiling continues for 24-30 hours and temperature is maintained by
cooling colls. Finally the fermentation takes place slowly and increase in alcohol is slow. Depending upon
the sugar contents present in the solution and temperature of the fermentation, the fermentation time
varies from 36-50 hours. The wash or wort is then distilled in coffey still.

DISTILLATION
The wash or wort is then distilled in a specially designed continuous still, called coffey still. It consists of
two tall fractionating columns, called the analyser and the rectifier, which are provided with perforated
horizontal plates having valves opening upwards. The coffey still works on the counter current principle
and steam and alcohol travel in opposite directions through the still.

The wash containing 6-10 percent alcohol is pumped to the upper sections of the still after passing several
heat exchangers. A current of steam is passed from the bottom end of the analyser. Steam passes up the
analyser and takes away the vapours of alcohol from the down coming wash. The mixture of alcohol and
steam vapours leaves the analyser at the top end and enters the rectifier near the base. Here, it heats the
wort flowing through the pipes on its way to the analyser. In the rectifier, condensation takes place and
most of the steam is condensed because of higher b.p. (100°C) than that of ethanol (78.3°C). The
uncondensed alcohol vapours escape near the top and are condensed with the help of a condenser. The
alcohol thus obtained is about 90% pure. The residue left in the still is used as cattle feed. 90% alcohol is
then subjected to fractional distillation. The first fraction contains low boiling liquids, such as
acetaldehyde. The second fraction contains 95% ethanol, called rectified spirit or industrial alcohol. and
last fraction contains fusel oil.

Industrial alcohol

Industrial alcohol is ethanol which is also called as ethyl alcohol, pure alcohol, grain alcohol, or drinking
alcohol. Ethanol is a volatile, flammable, colorless liquid.

Raw material

Corn

Water

Reactions:-
2(C6H10O5)n + nH2O (diastage) → nC12H22O11

C12H22O11 + H2O (maltase) → 2C6H12O6

C6H12O6 (zymase) → C2H5OH + 2CO2

Process Description
The corn is degerminated, dehulled and milled, either in wet or dry. The milled corn mass was conveyed
to the cooker. Cooking was gelatinized the ground grain so that the barley malt amylases can be converted
the starch to fermentable sugars. The cookers may be batch or continuous and are operated under
pressure.

In the continuous process the grain was precooked for 1-5min with water and stillage. The dealcoholized,
fermented beer was discharged from the bottom of the beer still. The mash was continuously fed to a
steam heater that raised the temperature to 1750C. The mash was passed through a series of pipes and
discharged through a relief valve into a flash chamber.

Time in the cooker was about 1.5min and pressure was maintained at 60-100kPa gages. The temperature
of the mash was dropped to about 600C in the flash chamber.

The water and malted barley mixed with gelatinized grain mash. The mix was pumped through pipeline
for 2min at 600C and then it was sent to fermenters through pipe coolers. The starch was hydrolyzed to
about 70% maltose and 30% dextrin in the short time in the converter. To lower the pH, providing nutrition
to yeast and provide buffering action stillage (20-25% of the final mash volume) from the beer still was.
The initial pH was adjusted to 4.8-5.0 with sulfuric acid and/or stillage. As the reaction indicated,
fermentation is exothermic, so cooling may be necessary to ensure that maximum temperature does not
exceed 320C.The time of the fermentation from 40-72hr.

The liquors in the fermenter, after the action was finished are called beer. The alco separated by
distillation. The beer containing 5.5-11% alcohol by volume was pumped to the upper section of the beer
still. As the beer passes down the column, it loses its lighter boiling constituents. The liquid discharged
from the bottom of the still through heat exchanger was known as Stillage. The overhead containing
alcohol, water, and aldehydes passes through heat exchanger to the partial condenser, or dephlegmator,
which condensed 50% alcohol, containing volatiles or aldehydes. The condensate was conducted into the
aldehyde column, from which the low boiling impurities were separated as an overhead. Te eluent liquor
from bottom of the aldehyde column flows into the tifying column. In this the Jerumn the alcohol was
brought to the strength and finally purified to 95-95.5%
ALCOHOL BEVERAGES

There are three types of alcoholic beverages-distilled liquors, (spirits) wines and beers. Spirits are obtained
by producing alcohol containing liquid by fermenting saccharified starch or beet sugar, figs, strawberrys
etc. and then distilling the liquid Rum, Gin, Whiskey, Brandy, Vodka etc. are well known spirits. In the case
of whiskey, the process of fermentation is same as described in the above paragraphs. Malty barley is
used for its production and the wash is distilled in a special pot still. As a result of it, a mixture of pure
alcohol and water is obtained, which also contains some fusel oil, aldehydes, esters etc. The raw whiskey
thus obtained is then placed in casks to mature. As a result, aldehydes are converted into acids by
oxidation. These acids react with alcohols present to form esters. These give a special flavour and aroma
to the whiskey.

Rum is prepared from sugar beet in France, carrot beet in Portugal, strawberry in Brazil, and Molasses in
Cuba. Rum prepared from fermented molasses, owes its flavour to the presence of ethyl acetate and ethyl
butyrate. The rum is also flavoured by placing sugar cane leaves in the still.

Gin is obtained from plum spirit. The spirit obtained by distillation is artificially flavoured by redis- tilling
with juniper berries.
Brandy is obtained by the distillation of wine. Brandies of various aroma are obtained by distilling different
fermented juice of fruits such as apple, peas, figs, melons etc. Vodka is a spirit or whiskey class obtained
in Russia from rye. Wines are prepared by the fermentation of fruit juices. Wines are classified as natural
(alcohol 7-14%), fortified (alcohol 14- 30%), sweet or dry, still or sparking. The chief juice is that of grapes.
The grape juice contains glucose, fructose and various acids, especially tartaric acid. The skin of grapes
contain tannin, various essential oils or it may also be a colouring matter. All these materials come into
the juice, when grapes are crushed in crusher. The latter macerates the grapes but does not crush the
seeds and also removes part of the steams. The juice extracted from the wooden roller. (called crushers)
is called must. The must is first fermented into ethyl alcohol. After first active fermentation is over, the
new wine is filled full into casks, which are loosely closed in order to prevent the conversion of alcohol
into acetic acid. A fermentation known as still fermentation is allowed to proceed in the casks for 3-4
months. During this period, the yeast settles down and tartaric acid, alongwith various salts and colouring
matters, separates out as argol. The clear wine is then allowed to ripe for 2-4 years. During ripening, tannin
and some other impurities are precipitated. Moreover, alcohol and fusel oil react with acids present to
form esters. The latter give characteristic flavour to the wine. After ripening, the wine is bottled. Since
quality of grapes varies from place, some vintages give better wines than others.

Beers of different varieties are obtained from malt. Malt is germinated barley. For making beer, killed malt
is used, in order to restrict the amount of diastase in the malt. Moreover, in the process of mashing
complete conversion of the starch into maltose is not allowed to take place. A portion of starch is
converted into dextrin by an enzyme (diastase is a mixture of enzymes).

(C6H10O5)n → (C6H10O5)20

That is why, the amount of diastase is restricted. The protein in the malt is converted into peptone by the
proteolytic enzymes present in the malt. The dextrin is retained inorder to give body to the beer. because
it escapes fermentation by yeast. Further more, the nitrogenous compounds, the albuminoids and
proteins in the grains are converted into peptones and other substances by the enzyme peptase present
in the malt. These also provide body to beer and increase the nutritive properties of beer. Nitrogenous
com- pounds and the phosphates supply food to the yeast, which is used to ferment the cold wort to
produce alcohol and carbondioxide.

When mashing is complete, the wort is withdrawn and boiled with hops and after settling, the clear liquid
is fermented with yeast. Yeast contains many enzymes, of these maltase and zymase take part in the
conversion of maltose into alcohol. Maltase hydrolyses maltose into glucose. while zymase converts
glucose into alcohol and CO2. After completion of the active fermentation, the new beer is passed into
casks and slow fermentation is allowed to take place. The froath formed is passed out through the bung
hole. The beer is drawn off into casks or bottles. When rennet (containing enzyme rennin) is added to
milk, it brings about curdling of the milk as a result of decomposition of casein present in the milk into
paracasein. The latter forms the curd and whey albumin.
In making cheese, a culture of bacteria capable of producing lactic acid from milk sugar is added to the
milk at a controlled temperature and then rennet is added. Temperature and acidity of the milk is carefully
controlled. As a result, casein separates out as a curd, from which whey is removed. The resulting curd is
allowed to mature for some time and then it is ground, salted and pressed in a cheese mould. The cheese
is then ripened. During ripening casein decomposes into a number of different substances in pres- ence
of enzymes. These decomposition products give characteristic flavour to the cheese.

In recent years the importance of micro biological processes has been fully recognised and their industrial
applications have been extended. Lactic acid, citric acid and various vitamins are manufactured 'from
fermentation process. These substances are of great importance in the production of antibiotics.

A polymer of glucose, called dextran has also been made recently by making use of bacterial activity. This
polymer is used therapeutically and as a serum substitute or expander in order to maintain the circulation
of blood volumes in cases of shock.

MANUFACTURE OF BEER
The steeped barley is first germinated and after the proper germination, the process of germination is
stopped by drying and curing as described earlier. As a result, killed malt is obtained. During the process
of malting the diastase formed converts only a part of starch into maltose.

Formation of wort - The raw grain of maize is first gelatinised with high pressure steam and then mixed
with malt grist and washed in mash tum at about 70°C with water. The mash tum is a circular cast iron or
copper vessel provided with a lid and rating machine inside for smashing the grist against the wall of the
cylinder. The mash tum is covered with lagging material for maintaining the temperature at about 60-
70°C. After mashing, the wort is washed with hot water and then the entire mass is run from the mash
tum to filter presses or straining tube, where the wort is separated from the spent grains through a slotted
false bottom and run into a copper vessel, where the wort is boiled or cooked and hopped. For complete
recovery of all substances in solution, a process, called sparging is used in which decarbonated water is
sprayed through the grains at 155°F. The boiling or cooking is done by passing steam through the Jacket
of the copper vessel and the purpose of boiling is to concentrate the wort of the desired strength, to
sterilize it and destroy all the enzymes, to coagulate certain proteins by heat (180°F), to modify the malty
smell of the wort, and to extract the hop resins, tannin and aroma from the hops. The latter are added
gradually during the cooking process. The best hops are added at the end of cooking in order to retain the
hop oil in the wort. 10-15 lbs of hops are sufficient per thousand gallons of the wort. The boiling takes
about 3 hours and then the spent hops are separated from the boiling wort very quickly through a false
bottom in the hop jack, a vessel made of cast iron or copper. The spent hops form a filtering layer on the
false bottom (provided with the hop jack), through which the wort is filtered from sedimentory matter.
The filtered wort is then pumped to a cooler made of cast iron or copper. Cooling not only reduces the
temperature of the wort, but also allows the wort to absorb enough air to facilitate the process of
fermentation. In the cooler, the proteins and hop resins are also precipitated. The hot wort may be first
cooled to about 150-160"F in a large shallow cooler and then in a horizontal, brine cooled copper tube of
the open Baudelot cooler. The wort from the cooler falls through the bottom of a perforated pan on the
copper pipes, through which cooled brine passes. The wort absorbs sufficient amount of air and is cooled
to about 13-14°C, the optimum temperature for fermentation.

Fermentation of wort - The cooled wort is then transferred to a fermentation unit, which consists of three
chambers, upper, middle and lower. The fermentation process is exothermic reaction and so temperature
tends to increase. The temperature is maintained by passing cold or hot water through the coils fitted
with all the chambers. The upper and middle chambers are fermentation chambers and lower chamber is
the yeast chamber. The middle chamber is also fitted with a skimmer and a parachute (a funnel shaped
apparatus). The top of parachute is flushed with the top of the middle chamber and end of its stem
extends to the lower yeast chamber.

First of all, yeast collected from previous operation is placed in smaller vessel and small amount of wort
is added. The yeast is allowed to ferment and grow. The yeast is then transferred to the upper chamber
of the fermentation unit and wort is allowed to run simultaneously. Whole wort is transferred and yeast
is allowed to grow for about 2 days. After completion of fermentation to some extent, the whole mass is
transferred to the middle chamber, where fermentation is allowed to proceed for 6-7 days. Al this state
fermentation becomes complete. The yeast which floats on the surface (top fermentation) is continuously
skimmed into the parachute, from where it enters the yeast chamber through the stem. The beer is then
collected and casked. The beer, which collects in the yeast chamber with the yeast, is separated from the
yeast by pressing. The separated beer is transferred to the cask. Slow fermentation is allowed to continue
in casks. At the conclusion of the process, the beer is drawn off into casks or bottles. The beer contains
about 3-6 percent alcohol. A part of the yeast collected in the yeast chamber is used for next operation
and rest is first removed from the bitter taste of hops and sold as edible yeast (a substitute for meat).

MANUFACTURE OF SPIRITS
Scotch whiskey is prepared from barley malt. The barley grains are dried, steeped with water for 2-4 days
and then germinated as already described in case of beer. In this case curing is carried out at such a low
temperature, which is sufficient enough for stopping the process of germination. In this manner, loss of
diastase is prevented. In the production of spirits from maize, potato, rye etc., the enzyme diastase is
provided by green malt. The latter is mixed with the starchy material after the process of gelatinisation.
In this case, long malt is used and can be prepared by steeping dry barley or wheat with 40-45% water for
2- 4 days in presence of dilute milk of lime, which prevents the growth of bacterial and moulds, because
green malt is not killed. The steeped barley is then allowed to germinate for about 20 days on the floor of
malt house in order to bring about the formation of maximum amount of diastase. When malt (barley) is
used, it is first ground to form grist. It is then mashed in the mash tum in presence of water as described
in case of beer. The mashed wort is then washed with hot water and then transferred into the
fermentation vat. In the case of spirits or distillery process, the mashing is carried out at least twice or
thrice. The wort, after mashing and washing with hot water is taken out by a pipe and passed into a vessel,
called underback. The wort is allowed to stand for some time in the vessel and the action of diastase is
thus allowed to continue.

If potato, maize, rye etc., are used as a source of starch, 4-5% green malt is mixed in case of potatoes and
9-10% in case of other grains, such as maize, rye etc. Before mixing the malt, the starchy materials are
gelatinised by high pressure steam for about 2-3 hours at 2 atmospheric pressure and increased to 4
atmospheres. The gelatinising is carried out in a cooker containing potatoes and water and pressure is
steam is allowed to pass through. Condensed water is removed from time to time. After steaming, the
pressure is released. As a result, super heated water and steam inside the potatoes suddenly expand and
hence gelatinised. The gelatinised starch, called mash is then cooled by dropping it on copper pipes cooled
with water or brine.

The mash cooled at SS'C is introduced in a vat and requisite amount of finely ground malt is added. The
mixture is kept at about 55°C for fifteen minutes, The mash is saccharified at this temperature and then
some more mash is introduced and temperature is regulated at 65°C. At this temperature, whole of the
starch gets liquefied. Now remaining hot mass is run into the wort vat and temperature is regulated at
67.7°C. At this temperature bacteria are completely killed. The diastase remains almost unaffected under
these conditions. This diastase alongwith yeast converts the dextrin into alcohol. The wort formation takes
about one hour. The lactic acid or hydrofluoric acid is added to sour the wort. The sour wort is cooled first
to 26.7°C and pitched with distillery yeast and again cooled to about 15.6°C slowly.

The temperature, however, increases to about 30°C by the heat of fermentation. This temperature is not
allowed to increase further. After adding mother yeast to a small portion of soured wort, the solution is
allowed to stand for 12-15 hours. During this time yeast grows rapidly and becomes ripe. The ripe yeast
thus formed is then added to the main soured wort, the temperature is maintained and fermentation is
allowed to continue for about 3 days.

MANUFACTURE OF WINES
Wines are prepared by the fermentation of fruit juices. Most important and widely employed is grape
juice, which contains glucose, fructose and various acids, especially tartaric acid. Skins of grapes contain
tannin, various essential oils and there may be colouring matter. The grapes are crushed mainly by
wooden rollers, called crushers. The crusher macerates them but does not crush the seeds and also
removes some of the stems. The juice extracted from the grapes is called must and contains glucose,
fructose, and other materials given above. The resulting must is run into large tanks, where sulphurous
acid is added to check the wild yeast. An active culture of selective and cultivated yeast (3-5% of the
volume of juice) is added and first active fermentation is allowed to proceed. The temperature is not
allowed to increase above 85°F by cooling coils, provided with the fermentation tanks. The CO2 gas is
liberated as a result of conversion of glucose into alcohol and this CO₂ carries the stems and seeds to the
top, which are partly prevented by a grating floated in the vat. This allows extraction of the colour and the
tannin from the skins and seed.
After the first active fermentation is over, the juice is pumped out of the bottom of the vat, back over the
top and the new wine is finally drawn into casks, which are filled full and loosely closed in order to prevent
the conversion of alcohol into acetic acid. In the casks, fermentation proceeds for several months, the
yeast ferments the remainder of the sugar. During this period, yeast settles down at the bottom and
tartaric acid along with various salts and colouring matters separates out. The wine is also given a celler
treatment during which the wine is first allowed to stand for about 2 months to remove suspended matter
and then racked for clarification. An insoluble precipitate with the tannin is also formed. Extra tannin may
also be added and then racked and filtered through asbestos or diatomaceous earth. The resulting
clarified wine is then blended with other wines, sugars, acid and tannin, in order to maintain commercial
standard.

Argols or potassium acid tartrate is removed from the wine by chilling and as a result, a more finished
wine is obtained.Argols or crude potassium acid tartrate constitute the commercial source of tartaric acid
and its compounds. Good sweet wine can also be obtained by a quick ageing method in about 4 months.
Pasteurization, refrigeration, sunlight. ultraviolet light, ozone, agitation and aeration are various ageing
methods. The wine may be kept at about freezing for 3-4 weeks and some oxygen is passed through it.
The wine is then racked, clarified and filtere

MANUFACTURE OF VINEGAR
Many weak alcoholic beverages, such as wines become sour when exposed to some time to the air. The
souring is due to the oxidation of alcohol to acetic acid by air in presence of certain fungi and bacteria,
e.g.. mycoderma aceti and bacterium aceti. This process is made use of in the production of vinegar.
Vinegar is a dilute solution of acetic acid and contains about 5.5 - 10% acetic acid. Vinegar stronger than
14% cannot be prepared, because bacteria causing the fermentation are killed.

Anaerobic oxidation of both ethanol and acetaldehyde is also possible by acetic bacteria in presence of
methylene blue, in which there is a direct participation of molecular oxygen in the hydrogen abstraction
process. This is much less likely during aerobic oxidation and it is very probable that a particular enzyme
system secreted by the bacteria is actually responsible for the oxidation of both aldehyde as well as the
alcohol, the oxygen acting as an aid for the bacterial cells to multiply fast in number and hence increasing
the concentration of the active enzyme (the real oxidising agent) rapidly. The most probable agent
bringing about the biodegradation has been found to be cytochrome oxidase or indophenol oxidase.

It should also be noted that temperature for fermentation should be 30-35°C, alcohol concentration
should be 12-14% (because in concentrated solutions the bacteria are killed), alcohol should be
contaminated with 10-12% acetic acid, because slightly sour media causes bacterial cell multiplication and
fer- mentation should be carried out in presence of nutrients such as MgSO4, (NH4)2SO4, (NH4)2HPO4

Moreover, addition of alcohol should be slow inorder to avoid increase in temperature, because the
process of fermentation or oxidation is exothermic. The supply or air during fermentation should also be
regulated because excess supply may cause the oxidation of acetic acid formed to CO2 and H2O by the
action of bacteria. Any fruit such as apple, grapes etc, 'containing more than 9% of sugar can be fermented
to vinegar in two steps. Moreover, sugar syrups, cornstarch, beer etc may also act as source of vinegar.
Vinegar is usually manufacture from malt, which is mashed and then fermented with diastase as in the
preparation of alcohol. The fruit extract or diluted molasses containing about 12% sugar is inoculated with
saccharomyces ellipsoids, a special wine yeast, which imparts a good flavour to fermented liquor and
ultimately goes to enrich the flavour of vinegar formed in the second phase of fermentation. The wort is
then fermented by yeast and the alcohol percentage is adjusted, if necessary. After fermentation, the
alcoholic liquor (containing less than 10% of alcohol) alongwith nitrogenous matter and salts necessary
for the growth of the bacteria is sprinkled over large wooden vat, provided with a perforated false bottom
and packed with basket work. Oxygen or air is admitted in the vat through the holes, provided at one side
of the vat near the bottom.