Table of content

1. Introduction
1.1
1.Introduction
1.1 General description of Ethanol
Production of industrial alcohol is an age old practice. But with time, the usage
areas as well as production techniques have gone through a major
transformation. Industrial alcohol is distilled ethyl alcohol(C2H5OH), normally of
high proof, produced and sold for other than beverage purposes. It is usually
distributed in the form of pure ethyl alcohol, completely denatured alcohol,
especially denatured alcohol and proprietary solvent blends. Ethyl Alcohol is the
common name for the hydroxyl derivative of the hydrocarbon ethane.
Industrial alcohol is distilled ethyl alcohol normally of high proof, produced and
sold for other than beverage purposes. Industrial alcohol finds its applications in
many chemical industries, pharmaceutical industries, Ink Industries and various
allied applications. Much of this alcohol is obtained synthetically from ethylene.
However, its production from microbial fermentation using variety of cheap
sugary substrates is still commercially important. The various substrates used for
ethanol production are sugar crops such as sugarcane, sugar beet, sorghum, etc.
provide a good substrate.
Bye product of these crop processing, e.g., molasses, sweet sorghum syrup, etc.
are the most common substrates. Cereals like maize, wheat, rice etc. areal soused
for ethanol production. Distillation of industrial alcohol, which is normally not
used for consumption, can be made in a two-step process. The process of
distillation is one with as low dynamics making it essential to have a carefully
planned and designed control system. Ethyl alcohol or ethanol ranks second only
to water as the most widely used solvent in chemical industry and as these
industries have expanded, so the demand for industrial alcohol has increased.
Table 2.1: Physical properties of Ethanol
Properties
Molecular formula C2H6O
Molar mass 46.07 g mol−1
Exact mass 46.041864814 g mol−1
Appearance Colorless liquid
Density 0.789 g/cm3
Melting point −114 °C, 159 K, -173 °F
Boiling point 78 °C, 351 K, 172 °F

1.2 History of product

The word alcohol derives from Arabic al-kuhul, which denotes a fine powder of
antimony used as an eye makeup. Alcohol originally referred to any fine powder,
but medieval chemists later applied the term to the refined products of
distillation, and this led to the current usage. Mostly, all the ethanol used
industrially is a mixture of 95% ethanol and 5% water, which is known simply as
95% alcohol. Although pure ethyl alcohol is available, it is much more expensive
and is used only when definitely required.
The world faces the fact that population increases vertiginously and
industrialization, as well as consumption of unrenewable resources such as oil and
its derivatives, grows in the same measure. To make economic growth
independent of the use of fossil fuels and environmental pollution from
combustion, interest has increased in using renewable resources to obtain
energy, known as alternative energy sources. A renewable resource is biomass
generated by photosynthetic organisms (autotrophs) that store energy in the
form of sugars, which can be transformed into ethanol for use as biofuel through
the process of fermentation. To reduce the competition for crop land that may
exist between renewable resources and food production, non-conventional (low-
cost) byproducts and raw material not used for human food or animal feed with a
high content of fermentable starch or sugars are sought.

The fermentation of sugar into ethanol is one of the earliest biotechnologies

employed by humans. Ethanol has historically been identified variously as spirit of
wine or ardent spirits, and as aqua vitae or aqua vita. The intoxicating effects of
its consumption have been known since ancient times. Ethanol has been used by
humans since prehistory as the intoxicating ingredient of alcoholic beverages.
Dried residue on 9,000-year-old pottery found in China suggests that Neolithic
people consumed alcoholic beverages.

The inflammable nature of the exhalations of wine was already known to ancient
natural philosophers such as Aristotle, Theophrastus, and Pliny the Elder.
However, this did not immediately lead to the isolation of ethanol, even despite
the development of more advanced distillation techniques in second- and third-
century Roman Egypt. An important recognition, first found in one of the writings
attributed to Jābir ibn Ḥayyān (ninth century CE), was that by adding salt to
boiling wine, which increases the wine's relative volatility, the flammability of the
resulting vapors may be enhanced. The distillation of wine is attested in Arabic
works attributed to al-Kindī and to al-Fārābī, and in the 28th book of al-Zahrāwī's
In the twelfth century, recipes for the production of aqua ardens ("burning
water", i.e., ethanol) by distilling wine with salt started to appear in a number of
Latin works, and by the end of the thirteenth century it had become a widely
known substance among Western European chemists.

The works of Taddeo Alderotti (1223–1296) describe a method for concentrating

ethanol involving repeated fractional distillation through a water-cooled still, by
which an ethanol purity of 90% could be obtained. The medicinal properties of
ethanol were studied by Arnald of Villanova (1240–1311 CE) and John of
Rupescissa (c. 1310–1366), the latter of whom regarded it as a life-preserving
substance able to prevent all diseases (the aqua vitae or "water of life", also
called by John the quintessence of wine).

In China, archaeological evidence indicates that the true distillation of alcohol

began during the Jin (1115–1234) or Southern Song (1127–1279) dynasties. A still
has been found at an archaeological site in Qinglong, Hebei, dating to the 12th
century. In India, the true distillation of alcohol was introduced from the Middle
East, and was in wide use in the Delhi Sultanate by the 14th century.

In 1796, German-Russian chemist Johann Tobias Lowitz obtained pure ethanol by

mixing partially purified ethanol (the alcohol-water azeotrope) with an excess of
anhydrous alkali and then distilling the mixture over low heat. French chemist
Antoine Lavoisier described ethanol as a compound of carbon, hydrogen, and
oxygen, and in 1807 Nicolas-Théodore de Saussure determined ethanol's
chemical formula. Fifty years later, Archibald Scott Couper published the
structural formula of ethanol. It was one of the first structural formulas
determined.

Ethanol was first prepared synthetically in 1825 by Michael Faraday. He found

that sulfuric acid could absorb large volumes of coal gas. He gave the resulting
solution to Henry Hennell, a British chemist, who found in 1826 that it contained
"sulphovinic acid" (ethyl hydrogen sulfate). In 1828, Hennell and the French
chemist Georges-Simon Serullas independently discovered that sulphovinic acid
could be decomposed into ethanol. Thus, in 1825 Faraday had unwittingly
discovered that ethanol could be produced from ethylene (a component of coal
gas) by acid-catalyzed hydration, a process similar to current industrial ethanol
synthesis.

Ethanol was used as lamp fuel in the U.S. as early as 1840, but a tax levied on
industrial alcohol during the Civil War made this use uneconomical. The tax was
repealed in 1906. Use as an automotive fuel dates back to 1908, with the Ford
Model T able to run on petrol (gasoline) or ethanol. It fuels some spirit lamps.
Ethanol intended for industrial use is often produced from ethylene. Ethanol has
widespread use as a solvent of substances intended for human contact or
consumption, including scents, flavorings, colorings, and medicines. In chemistry,
it is both a solvent and a feedstock for the synthesis of other products. It has a
long history as a fuel for heat and light, and more recently as a fuel for internal
combustion engines.

1.3 objective of the product

1. Medical

A. Anesthetic

Ethanol is the oldest known sedative, used as an oral general anesthetic during
surgery in ancient Mesopotamia and in medieval times. Mild intoxication starts at
a blood alcohol concentration of 0.03-0.05% and induces anesthetic coma at
0.4%. However, this use carried the high risk of deadly alcohol intoxication and
pulmonary aspiration on vomit, which led to use of alternatives in antiquity, such
as opium, cannabis, and later diethyl ether in the late Middle Ages.

B. Antiseptic

Ethanol is used in medical wipes and most commonly in antibacterial hand

sanitizer gels as an antiseptic for its bactericidal and anti-fungal effects. Ethanol
kills microorganisms by dissolving their membrane lipid bilayer and denaturing
their proteins, and is effective against most bacteria, fungi and viruses. However,
it is ineffective against bacterial spores, but that can be alleviated by using
hydrogen peroxide. A solution of 70% ethanol is more effective than pure ethanol
because ethanol relies on water molecules for optimal antimicrobial activity.
Absolute ethanol may inactivate microbes without destroying them because the
alcohol is unable to fully permeate the microbe's membrane. Ethanol can also be
used as a disinfectant and antiseptic because it causes cell dehydration by
disrupting the osmotic balance across the cell membrane, so water leaves the cell
leading to cell death.

C. Antidote
Ethanol may be administered as an antidote to ethylene glycol poisoning and
methanol poisoning. Ethanol serves this process by acting as a competitive
inhibitor against methanol and ethylene glycol for alcohol dehydrogenase.
Though it has more side effects, ethanol is less expensive and more readily
available than fomepizole, which is also used as an antidote for methanol and
ethylene glycol poisoning.

D. Medicinal solvent

Ethanol, often in high concentrations, is used to dissolve many water-insoluble

medications and related compounds. Liquid preparations of pain medications,
cough and cold medicines, and mouth washes, for example, may contain up to
25% ethanol and may need to be avoided in individuals with adverse reactions to
ethanol such as alcohol-induced respiratory reactions. Ethanol is present mainly
as an antimicrobial preservative in over 700 liquid preparations of medicine
including acetaminophen, iron supplements, ranitidine, furosemide, mannitol,
phenobarbital, trimethoprim/sulfamethoxazole and over-the-counter cough
medicine.

E. Pharmacology

In mammals, ethanol is primarily metabolized in the liver and stomach by alcohol

dehydrogenase (ADH) enzymes. These enzymes catalyze the oxidation of ethanol
into acetaldehyde (ethanal):

CH3CH2OH + NAD+ → CH3CHO + NADH + H+

When present in significant concentrations, this metabolism of ethanol is
additionally aided by the cytochrome P450 enzyme CYP2E1 in humans, while
trace amounts are also metabolized by catalase.

The resulting intermediate, acetaldehyde, is a known carcinogen, and poses

significantly greater toxicity in humans than ethanol itself. Many of the symptoms
typically associated with alcohol intoxication—as well as many of the health
hazards typically associated with the long-term consumption of ethanol—can be
attributed to acetaldehyde toxicity in humans.
The subsequent oxidation of acetaldehyde into acetate is performed by aldehyde
dehydrogenase (ALDH) enzymes. A mutation in the ALDH2 gene that encodes for
an inactive or dysfunctional form of this enzyme affects roughly 50% of east Asian
populations, contributing to the characteristic alcohol flush reaction that can
cause temporary reddening of the skin as well as a number of related, and often
unpleasant, symptoms of acetaldehyde toxicity. This mutation is typically
accompanied by another mutation in the alcohol dehydrogenase enzyme ADH1B
in roughly 80% of east Asians, which improves the catalytic efficiency of
converting ethanol into acetaldehyde.

2. Fuel

The largest single use of ethanol is as an engine fuel and fuel additive. Brazil in
particular relies heavily upon the use of ethanol as an engine fuel, due in part to
its role as one of the world's leading producers of ethanol. Gasoline sold in Brazil
contains at least 25% anhydrous ethanol. Hydrous ethanol (about 95% ethanol
and 5% water) can be used as fuel in more than 90% of new gasoline-fueled cars
sold in the country.

The US and many other countries primarily use E10 (10% ethanol, sometimes
known as gasohol) and E85 (85% ethanol) ethanol/gasoline mixtures. Over time, it
is believed that a material portion of the ≈150-billion-US-gallon (570,000,000 m3)
per year market for gasoline will begin to be replaced with fuel ethanol.

Australian law limits the use of pure ethanol from sugarcane waste to 10% in
automobiles. Older cars (and vintage cars designed to use a slower burning fuel)
should have the engine valves upgraded or replaced.

According to an industry advocacy group, ethanol as a fuel reduces harmful

tailpipe emissions of carbon monoxide, particulate matter, oxides of nitrogen, and
other ozone-forming pollutants. Argonne National Laboratory analyzed
greenhouse gas emissions of many different engine and fuel combinations, and
found that biodiesel/petrodiesel blend (B20) showed a reduction of 8%,
conventional E85 ethanol blend a reduction of 17% and cellulosic ethanol 64%,
compared with pure gasoline. Ethanol has a much greater research octane
number (RON) than gasoline, meaning it is less prone to pre-ignition, allowing for
better ignition advance which means more torque, and efficiency in addition to
the lower carbon emissions.

Ethanol combustion in an internal combustion engine yields many of the products

of incomplete combustion produced by gasoline and significantly larger amounts
of formaldehyde and related species such as acetaldehyde. This leads to a
significantly larger photochemical reactivity and more ground level ozone. This
data has been assembled into The Clean Fuels Report comparison of fuel
emissions and show that ethanol exhaust generates 2.14 times as much ozone as
gasoline exhaust. When this is added into the custom Localized Pollution Index
(LPI) of The Clean Fuels Report, the local pollution of ethanol (pollution that
contributes to smog) is rated 1.7, where gasoline is 1.0 and higher numbers
signify greater pollution. The California Air Resources Board formalized this issue
in 2008 by recognizing control standards for formaldehydes as an emissions
control group, much like the conventional NOx and Reactive Organic Gases
(ROGs).

More than 20% of Brazilian cars are able to use 100% ethanol as fuel, which
includes ethanol-only engines and flex-fuel engines. Flex-fuel engines in Brazil are
able to work with all ethanol, all gasoline or any mixture of both. In the U.S. flex-
fuel vehicles can run on 0% to 85% ethanol (15% gasoline) since higher ethanol
blends are not yet allowed or efficient. Brazil supports this fleet of ethanol-
burning automobiles with large national infrastructure that produces ethanol
from domestically grown sugarcane.

Ethanol's high miscibility with water makes it unsuitable for shipping through
modern pipelines like liquid hydrocarbons. Mechanics have seen increased cases
of damage to small engines (in particular, the carburetor) and attribute the
damage to the increased water retention by ethanol in fuel.

A. Rocket fuel

Ethanol was commonly used as fuel in early bipropellant rocket (liquid-propelled)

vehicles, in conjunction with an oxidizer such as liquid oxygen. The German A-4
ballistic rocket of World War II (better known by its propaganda name V-2), which
is credited as having begun the space age, used ethanol as the main constituent
of B-Stoff. Under such nomenclature, the ethanol was mixed with 25% water to
reduce the combustion chamber temperature. The V-2's design team helped
develop U.S. rockets following World War II, including the ethanol-fueled
Redstone rocket which launched the first U.S. satellite. Alcohols fell into general
disuse as more energy-dense rocket fuels were developed, although ethanol is
currently used in lightweight rocket-powered racing aircraft.

B. Fuel cells

Commercial fuel cells operate on reformed natural gas, hydrogen or methanol.

Ethanol is an attractive alternative due to its wide availability, low cost, high
purity and low toxicity. There is a wide range of fuel cell concepts that have
entered trials including direct-ethanol fuel cells, auto-thermal reforming systems
and thermally integrated systems. The majority of work is being conducted at a
research level although there are a number of organizations at the beginning of
the commercialization of ethanol fuel cells.

C. Household heating and cooking

Ethanol fireplaces can be used for home heating or for decoration. Ethanol can
also be used as stove fuel for cooking.

3. Feedstock

Ethanol is an important industrial ingredient. It has widespread use as a precursor

for other organic compounds such as ethyl halides, ethyl esters, diethyl ether,
acetic acid, and ethyl amines.

4. Solvent

Ethanol is considered a universal solvent, as its molecular structure allows for the
dissolving of both polar, hydrophilic and nonpolar, hydrophobic compounds. As
ethanol also has a low boiling point, it is easy to remove from a solution that has
been used to dissolve other compounds, making it a popular extracting agent for
botanical oils. Cannabis oil extraction methods often use ethanol as an extraction
solvent, and also as a post-processing solvent to remove oils, waxes, and
chlorophyll from solution in a process known as winterization.
Ethanol is found in paints, tinctures, markers, and personal care products such as
mouthwashes, perfumes and deodorants. However, polysaccharides precipitate
from aqueous solution in the presence of alcohol, and ethanol precipitation is
used for this reason in the purification of DNA and RNA.

5. Low-temperature liquid

Because of its low freezing point of −114 °C (−173 °F) and low toxicity, ethanol is
sometimes used in laboratories (with dry ice or other coolants) as a cooling bath
to keep vessels at temperatures below the freezing point of water. For the same
reason, it is also used as the active fluid in alcohol thermometers.

2. Main raw material of Industrial alcohol

2.1 Available raw materials
Most of the ethanol is produced from renewable raw materials, which are
essentially sources of carbohydrates as sugars (sugarcane, beet, sweet sorghum,
Jerusalem arti- choke, fruits), starch (corn, wheat, cassava, sweet potato), and
lignocellulosic materials.

The ethanol industry of today utilizes raw materials rich in saccharides,

such as sugar cane or sugar beets, and raw materials rich in starch, such as corn
and wheat. The concern about supply of liquid transportation fuels, together with
the concern about global warming, have turned the interest towards large-scale
ethanol production from lignocellulosic materials, such as agriculture and forestry
residues. Baker’s yeast Saccharomyces cerevisiae is the preferred fermenting
microorganism for ethanol production because of its superior and well-
documented industrial performance. Extensive work has been made to
genetically improve S. cerevisiae to enable fermentation of lignocellulosic raw
materials. Ethanolic fermentation processes are conducted in batch, fed-batch, or
continuous mode, with or without cell recycling, the relative merit of which will
be discussed.
First generation Alcohol is obtained by the process of fermentation of the
sugars present in the different raw materials, which can be metabolized by
microorganisms of the genera Saccharomyces, Zymomonas, Kluyveromyces, and
Zygosaccharomyces.

Raw materials with available fermentable sugars

These sugars are present in:-

 sugarcane,
 sugar beets and
 sweet sorghum.

Use of these raw materials requires only milling, fermentation,

distillation and denaturalization (so that it is not apt for human
consumption). Moreover, for its use in mixtures with gasoline, it must
be dehydrated, which is necessary to displace its azeotropic point.
Methods of dehydration include adsorption with molecular meshes,
pervaporation, vacuum distillation, extractive distillation with solvents
or with salts or with both at the same time. Ethanol is also dehydrated
with hybrid processes that combine two or more of the above
processes.

In Brazil, they are researching how to reduce ethanol production costs

through integration in the use of energy and systems of co-generation
in production plants and decreasing expenditures of vapor and water,
or the use of sugarcane bagasse in the integration of first generation
(1G) and second generation (2G) ethanol production processes; 2G
processes involve the use of lignocellulosic substrates (as will be
discussed below) and the use of sweet sorghum as complementary raw
material, grown in areas neighboring the sugarcane and harvested
between cycles of the main crop (Jonker et al., 2015). Also studied is the
sustainable development and updating of the technologies of
biorefineries for producing bioelectricity (Khatiwada et al., 2016). After more
than 50 years of experience with 1G ethanol, progress was oriented
more toward optimization of the integral use of raw materials and the
byproducts of the processes, integration of first generation processes
with second generation processes, analysis of the effects of the
varieties of the same raw material, crop conditions, harvest seasons
and methods, and storage time of the raw material on ethanol yields.
One important aspect is mechanized harvest of sugarcane, which
increases the presence of inorganic compounds such as potassium,
calcium, silica, iron and copper, relative to the traditional process of
burning and manual cutting. report increases of 13 % calcium, 32 %
magnesium and 7.6 % silica in the sugarcane juice after mechanical
harvesting, relative to traditional burning and cutting. These minerals
affect fermentation because magnesium increases ethanol yield, but an
excess of copper reduces yields to 0.35 g g-1.

Interest in sweet sorghum and other sorghum types for ethanol

production has increased. This crop can be grown with little water
(compared with other cereals) and is highly efficient photosynthetically
for conversion of CO2 to biomass. Moreover, total sugar content of the
juice of the sweet sorghum stem is comparable with that of sugarcane
juice. Sugarcane processing machinery has been adapted to harvest and
mill sweet sorghum. proposed designing machinery specifically for
sorghum stems to improve juice extraction indexes. Sorghum has many
qualities, but its high moisture content (70%) and the low bulk density
limit the distances within which transporting it after harvest is
profitable. Also, conservation of the sugars in the stems is difficult
because the native bacteria of the crop, such as Leuconostoc, degrade a
large proportion of sugars during transport and storage. Inhibitors of
Leuconostoc growth during transport, such as SO2 gas, have been
tested. However, storing the gas in hermetic tanks at the sorghum
collection sites, as well as its elimination when it arrives at the
processing plant, make it an expensive system.
Extracted sweet juices lose 20 % of their fermentable sugar content in
three days at ambient temperature and 40 % to 50 % in seven days.
Other affected aspects are decreased pH, increase in total acidity and
quantity of reducing sugars. Furthermore, the presence of ethyl
acetate, acetaldehyde and ethyl carbamate in sweet juices have a
negative effect on fermentation, causing low yields. Juices from milled
material must be filtered, clarified and pasteurized. Filtering removes
lignocellulosic fractions, leaves, dirt and other contaminants from the
collection stage. Clarifying is done to eliminate inorganic compounds,
and pasteurization eliminates the microorganisms present. For
clarification, the processes of sedimentation, sulfation, and
carbonatation generate byproducts known as sludge. To generate less
contaminating sludge from clarification, natural products are used, such
as extracts from leaves and seeds of Moringa oleifera Lam., which
remove more iron and calcium than synthetic polymers. High-pressure
treatment (600 MPa 6 min-1) is an alternative to high temperatures for
elimination of microorganisms and inactivating enzymes from sweet
juices. Also, ohmic heating for 1 min and 32 V cm-1 has been tested to
inactivate polyphenol oxidase and reduce microorganisms. High-
pressure methods and ohmic heating maintain juice quality stable and
inhibit enzymatic activity, thus increasing its useful life in storage.
Therefore, the search for methods of conserving raw materials and
sweet juices for long periods without loss of fermentable sugars
requires further study.

First generation technology has been applied industrially for 40 years.

For this reason, besides the studies mentioned, there are many studies
related to analysis of environmental problems this technology
generates. Among these problems is the biomass produced, separated
in the process of distillation and known as vinasse, which is produced at
a ratio of 12 L vinasse L-1 ethanol. Vinasse is a liquid with suspended
particles, acid pH and high chemical oxygen demand (COD) because of
the organic matter present. Forty years ago, vinasse was a highly
contaminating liquid residue that caused serious environmental
problems in the aquifers where it was discharged. Its final disposal was
a problem because there was little technology to treat it and costs were
high. This situation changed because of new technologies with better
results that decreased operation costs. To treat residues of alcoholic
fermentation, physicochemical, chemical and biological methods were
tested. The biological methods are the most appropriate due to the
large quantity of organic biodegradable compounds present in their
composition. Several alternatives were proposed for the treatment or
reuse of vinasses, among which is feeding chickens, production of
biogas, and fertigation. In addition, standardized methods were created
to analyze life cycles to determine entrance and exits of energy,
evaluate generation of byproducts and residues in the entire supply
chain, and minimize impacts from crop establishment to consumption
of fuel ethanol (Cavalett et al., 2012; Gallejones et al., 2015; Miret et al., 2016).

From the above, it can be inferred that the main interests concerning
raw materials with high fermentable sugar content are increasing
yields, improving energy balances, diversifying raw materials, and
diminishing environmental impact.

Raw materials with high starch content

Other first-generation sources used to obtain ethanol are raw materials

that have a high starch content, such as cereals, tubers and rhizomes.
For these raw materials, the following steps are used for ethanol
production: milling, liquification, saccharification, fermentation,
distillation and dehydration. The country with more published studies
on ethanol from starch between 2012 and 2015 was China, the US and
India with 187, 181 and 74 published documents, respectively. The
main research topics center in three fundamental areas: understanding
agronomic aspects and their effect on ethanol yields, new operations in
industrial processes, and new laboratory techniques to predict starch
contents and ethanol yields.

In agronomy, the study of maize and sorghum with several focuses is

highlighted, especially on final ethanol yield, for example effects of
deficient irrigation (Liu et al., 2013), content of moisture at harvest (Huang et al.,
2012
), fungus infections and harvest maturity (Dien et al., 2012). In other
cereals, transgenic rice with high carbohydrate content was analyzed to
evaluate its ethanol yields (Kim et al., 2015).

In industry, the method most used to fragment the cereals is dry milling
(Orts and McMahan 2016), in which whole cereal grains are converted into flour,
which is processed without separating the grain components. The flour
is mixed with water to form a paste and α-amylase, pullulanase and
amyl glycosidase are added to convert the starch into glucose.
Ammonia is added to the process as a nitrogen source for the yeast and
to control system pH. This high-glucose mixture is converted to ethanol
by action of the yeast, while the solid residue, which has high fiber, lipid
and protein contents, is toasted and converted into a byproduct known
as distillers degraded grain soluble (DDGS). DDGS are mixed with syrup
from the process and sold as a supplement for livestock. Wet milling is
less used because of its high cost and water consumption, but starch
from the grains is isolated and ethanol yield is higher (Kandil et al., 2011). Dry
milling begins the process with starch, which is mixed with lipids and
proteins, and in the processes of liquefication and saccharification
these macromolecules decrease enzyme activity because they act as
inhibitors of the process (Srichuwong et al., 2010). For this reason, we
recommend investigating the interactions of the non-starch
components (lipids and proteins) with starch and their effects during
hydrolysis. To understand whether pigments are also inhibitors of the
process Wang et al. (2016) validated the use of pigmented sorghum and
determined that the high content of anthocyanins had no effect on
ethanol yield.
In the industrial stage there are innovative methods for extracting
fermentable sugars and non-structured carbohydrates with a diffusion
process that uses biomass from chopped sweet sorghum stems and
grains (Appiah-Nkansah et al., 2016). The process of milling and elimination of
physical barriers that limit interaction of enzymes with the starch was
studied and, according to Chuck et al. (2012), ethanol yield improves when the
sorghum grains are decorticated. In the studies mentioned, cereal
treatment technologies and ethanol yield are compared; energy
expenditures decrease and ethanol yields increase. However,
quantitative analysis of expenditures and comparison of different
process proposals should be studied. It is also necessary to evaluate
water balances and economic analyses of the process before initiating
industrial scaling.

In the laboratory, development of a practical method of detecting

approximate starch and ethanol contents using spectroscopy FT-NIR is
highlighted (Li et al., 2015). This could speed up industrial scale analysis to
control the ethanol production process systematically.

Fruits, rhizomes, roots and tubers are non-conventional sources of

starch used particularly in tropical countries. Graefe et al. (2011) analyzed
ethanol production from bananas in Costa Rica and Ecuador. Bananas
have an advantage over tubers because of their ripening and, because
they are climacteric, they naturally hydrolyze starch and thus no
enzyme treatment is required to reach the fermentable sugars (Asiedu, 1987;
Bugaud et al., 2009
). Production of ethanol from bananas at different stages of
ripeness showed that the highest ethanol yield was from green
bananas, immediately after harvesting. Ethanol yield from overripe
bananas was 23 % less than with immature bananas. This result is
attributed to a decrease in dry-base matter because of the metabolic
activity during ripening (Hammond et al., 1996). Due the higher yields, it is
recommended to used green bananas to produce ethanol. However,
technical problems exist, such as peeling the fruit because the shape of
the bananas makes automatizing the process difficult, and the peel is
strongly attached to the pulp at that physiological stage of the fruit.

One problem of biotechnological processes is inhibition by the end-

product. In the measure that alcoholic fermentation occurs, ethanol is
produced, which inhibits the metabolic pathway and the process of
bioconversion is detained until it causes cell death. A method used to
decrease this inhibition is to ferment the sugars immediately after their
release. This is achieved by simultaneous saccharification and
fermentation (SSF). Fermentable sugars are produced by starch
hydrolysis, and fermentation is carried out in the same reactor. Several
raw materials were processed in this way, and the results are better
than separate saccharification and fermentation. With green bananas,
theoretic ethanol efficiencies above 95 % were obtained in an enzyme
SSF system and the use of glucoamylases, pectinolytic enzymes and
yeast (Bello et al., 2012). Feasibility of a non-domesticated variety of mandioca
(Manihot glaziovii), as a non-food raw material with a high starch
content, was evaluated for production of ethanol by SSF. Ethanol
concentrations of 190 g L-1 were obtained. Another technological
development in inhibitors is Zimomonas mobilis bacteria isolated from
the African palm and Mexican pulque. They are osmotolerant and
resistant to high concentrations of ethanol and permit fermentation at
high concentrations, or very high gravity (VHG). The advantage is that
at high sugar concentrations (up to 360 g L-1) there is no contamination
by other bacteria. In these conditions, ethanol concentration from
sweet sorghum was 17.6% v/v (Deesuth et al., 2016). Process integration can
produce better yields than individual processes. Chu-ky et al. (2016) tested
simultaneous very high gravity liquefication, saccharification and
fermentation (SLSF-VHG) in rice by-products (broken rice grain) and
obtained a theoretical 83.2% ethanol.

In Brazil and Colombia new sources and methods for ethanol

production are being sought. Soccol (1997) achieved degradation of raw
mandioca starch granules during fermentation in solid state with
amyloglucosidase from Rhizopus oryzae.Cinelli et al. (2015) described the
method of cold hydrolysis of starches in which the process’ energy
demand decreases because of starch hydrolysis at temperatures below
its gelling temperature, that is, with starch in its granular form. This
technology generates another perspective of the process as an
alternative method that can be carried out without the liquefication
stage, which is achieved through amylolytic enzyme complexes
composed of endo-amylases, exo-amylases and debranching enzymes.
In Indonesia, Jusuf and Ginting (2014) and Kusmiyati (2015) studied sweet potato
(Ipomoea batatas L.) and the tuber Iles Iles (Amorphophalus
campanulatus), respectively, as sources for ethanol production. These
tubers have high potential for transformation because of their high
yield per unit of cultivated area. Sweet potatoes yield more than 30 t
ha-1 of tubers, which are 22.5 % starch, wet base, while Iles Iles has the
advantage of being cheap and not used for human food. As an
alternative source, ethanol yield of starch extracted from Taro corm
(Colocasia esculenta), a perennial plant of the Araceae family was
evaluated. Taro is cultivated in the tropics and subtropics of South
America, Asia, Oceania and Africa (Wu et al., 2016). With this raw material,
theoretic yields of 94.2% ethanol were achieved, an attractive yield for
ethanol production.

Second generation

In the last decade, most of the research in liquid fuels evaluated the
commercialization of ethanol produced with farm residues (cereal
straw, leaves and dry branches of forest crops) or industrial residues
(sugarcane bagasse and DDGS), composed mainly of cellulose,
hemicellulose and lignin; these biofuels are 2G.

The amount of residue generated by an industry and that can be used

to produce 2G ethanol is illustrated by Brazilian ethanol production
units, which processed more than 602 x106 t of sugarcane and
produced 24 x 109 L of ethanol from 2013 to 2014. Each ton of
sugarcane processed generates 270 to 280 kg of bagasse (Canilha et al., 2012),
demonstrating the high potential of residual biomass for 2G ethanol
production with these lignocellulosic residues.

These residues are the source for cogeneration of heat and electricity in
distilleries through thermochemical processes, such as direct
combustion, pyrolysis or gasification (Henrique et al., 2014), but their
biochemical transformation has certain technical limitations due to the
chemical nature of the lignocellulosic residues, which is heterogeneous
and different pretreatments, or modifications to the raw material, must
be applied. It is necessary to conduct laboratory studies that show the
technical feasibility of bioconversion of lignocellulosic residues to
ethanol.

Enzyme hydrolysis of cellulose is a limiting step in 2G ethanol

production because it needs complex enzymatic cocktails to de-
polymerize the residues, in addition to the complexity of the structural
arrangement of the components of the lignocellulosic residues that
make hydrolysis difficult. Lignin is a main component of these residues,
forming a structural arrangement together with the cellulose and the
hemicellulose that makes it difficult for cellulases to access the
cellulose hydrolyze it to glucose. Moreover, some cellulolytic enzymes
are often adsorbed by the hydrophobic surface of lignin that, therefore,
prevents them from acting on cellulose (Huron et al., 2016). Phenolic
compounds (tannins) of lignin, when de-polymerized, also become
inhibitors of fermentation, so that the effects and interactions of the
inhibitors of the processes of fermentation should be analyzed and
understood to enable improvement of production yields.

As mentioned in the section about the first generation, green or

immature banana pulp can be used to produce ethanol, but one of the
limiting steps is the peeling process. Eliminating the banana peel can be
avoided because both the banana peel and the pulp have high
carbohydrate content (Gebregergs et al., 2016), and both parts have been
studied for ethanol production (Parthiban et al., 2011). Therefore, ethanol can
be obtained from unpeeled green bananas. However, it should be taken
into account that some carbohydrates in the pulp and peel of green
bananas are complex polysaccharides and cannot be metabolized by
yeasts. For this reason, it is necessary to hydrolyze the lignin, the
cellulose and pectin to convert them into simple sugars. This is an
example of a raw material for ethanol production found in 1G and 2G
groups.

Milling bananas with their peel has technical problems derived from the
high viscosity and presence of fibers during and after milling (Afanador, 2005).
Oberoi et al. (2011)
report the use of dryers such as electric ovens to obtain a
dry brittle material that can be ground in laboratory mills, but in
industrial processing this would increase energy costs. Here, there is a
knowledge gap regarding decreasing viscosity in this step of the process
without drying and milling or the use of alternative energy (solar
energy, for example) for the drying process.

Second generation technology has given way to the concept of

biorefineries because through 2G processes ethanol is obtained as well
as methanol, synthesis gas, 2.5-dimethylfurane and tannins, a
determining factor for obtaining diverse products from lignocellulosic
raw materials. In this sense, one of the goals is to integrate 1G and 2G
technologies. Flório and Junior (2013) performed a heat analysis of production of
electricity and 2G ethanol from sugarcane bagasse and obtained better
heat effectivity since, besides fermenting sugarcane sucrose and the
glucoses that constitute cellulose, the process was optimized, and they
sought to ferment pentoses (mainly xylose) that form a structural part
of the hemicellulose. The process of bioethanol production was
simulated with the mixture glucose/xylose, and the effect of
temperature, pH and sugar concentrations was evaluated. The variables
of the process were reproduced using different conditions of operation
and it was demonstrated that simulation is a useful tool for generating
an optimal profile of alcoholic fermentation with the glucose/xylose
mixture (Reyes et al., 2016).

With the concept of biorefinery, Xu et al. (2018) propose obtaining diverse

products from lignocellulosic raw material using a modified method of
simultaneous saccharification and fermentation (MSSF), which involves
ecological technologies like hydrothermal pre-treatments and organic
solvents for production of a high cellulose content solid to be
transformed into ethanol. In addition, solvents are recycled to produce
adhesives based on plant proteins, and the xylose present in the
aqueous phase is used to obtain furfural. This type of integrated
proposals may be attractive for promoting commercial ethanol
production from lignocellulosic biomass.

Saccharomyces cereviciae and Zymomonas mobilis, used to ferment

hexoses, cannot use pentoses as a substrate because the metabolic
route for pentose conversion is unlike that of hexoses. For this reason,
other yeasts and bacteria are being studied. Candida shehatae (Guan et al.,
2013
) and Pichia stipitis (Travaini et al., 2016) have a high potential for fermenting
pentoses; their disadvantage is their low tolerance to ethanol, and in
the processes with these microorganisms inhibition by the end-product
occurs. As a solution to this technical problem, Kluveromyces
marxianus yeast has been proposed; it tolerates ethanol and can
ferment hexoses and pentoses (Lin et al., 2013). To increase use of pentoses,
process integration through use of fungi that metabolize wood without
pre-treating the lignin is being investigated. Mattila et al. (2017) used a single
step and a single microorganism for production of bioethanol with
phlebioid fungus species, achieving an ethanol yield of 5.9 g L-1.

Third generation
The search for raw materials for ethanol production has increased
interest in rapid generation of biomass with high energy density
(energy crops). Among these crops are found perennial grasses, micro-
and macro-algae, and cyanobacteria, all grouped in the denomination
of third generation (3G) biofuels. Algae are not seen as raw material for
ethanol production, but as producers of hydrogen as the substrate for
thermochemical conversion and as lipids for biodiesel (Brennan and Owende
2010
). Starches can be obtained by converting biomass from macroalgae.
The starches are then hydrolyzed and fermented to produce bioethanol
(Adams et al., 2009; Khambhaty et al., 2013; Scholz et al., 2013; Sudhakar et al., 2016) and biobutanol,
which has a higher energy density and greater compatibility with
gasoline than bioethanol (Dürre, 2008). One study on the economic
feasibility of microalgae as raw material for biorefineries was reported
by Konda et al. (2015). They highlight the potential advantages of producing a
broad portfolio of chemical products from microalgae to give economic
viability to industrial clusters denominated biorefineries. In the
laboratory, isolated alga strains are handled, but in natural conditions
water sources have a problem called eutrophication, which is the
proliferation of algae because of excess nutrients in the water. To solve
this problem, using these algae was proposed for ethanol production
and water quality improvement. Chen et al. (2017) patented a pre-treatment
technique using electro-coagulation and acid saccharification of algae in
lakes. They obtained up to 156 mg glucose g-1 of algae, which is
available for transformation to ethanol. The type of macroalgae most
used industrially are seaweeds, which are pluricellular organisms that
efficiently convert nutrients in the water and CO2 into biomass. There
are around 9,200 species of seaweed, but only 221 are economically
important (Mohammed, 2013). Macroalgae are raw material for bioethanol
production because they do not have a high content of lipids, as do
microalgae, but they do have a high content of sugars and other
carbohydrates that can be fermented. The alga growth variables
studied are those that affect production of carbohydrates, such as light,
temperature, nutrients, salinity and pH. With respect to the effect of
light, according to George et al. (2014), a photosynthetic light intensity of 60
μmol m-1s-1 and 12:12 h (light:dark) cycles achieved a biomass
production of 7.9 mg L-1 d-1 with Ankistrodesmus falcatus. It is important
to consider that the pretreatments, liquefication and saccharification of
the macroalgae have difficulties similar to those of the second-
generation raw materials since they also contained polysaccharides
that must be hydrolyzed to fermentable sugars, an aspect that requires
more research. Microalgae have a high content of lipids, but also of
carbohydrates. For this reason, Scenedesmus sp. has been proposed as
raw material for integrated production of ethanol and biodiesel
(Sivaramakrishnan and Incharoensakdi, 2018).

Another high-density crop in lakes and lagoons is water hyacinth

(Eichhornia crassipe), which is an environmental problem because of its
high growth rate; it prevents light and oxygen from penetrating these
bodies of water and cause the death of other species in these
ecosystems. Water hyacinth is composed mainly of cellulose, which can
be converted to glucose, the fermentable substrate for ethanol
production (Zhang et al., 2018). Different pre-treatments were tested for water
hyacinth, and the most effective was the combination of diluted acid
and microorganisms (Phanerochaete chrysosporium), which resulted in
highly purified cellulose (39.4 %) and high content of reducing sugars
(430.66 mg g−1).

Fourth generation

Advances in bioengineering have led to the concept of fourth

generation biofuels (4G), which use genetically modified organisms
(GMO) that capture more CO2, such as genetically modified sugarcane
that has high content of lipids for simultaneous production of ethanol
and biodiesel (Huang et al., 2016). Also, fermentation with genetically modified
E. coli produces triglycerides from sugarcane sweet juices that are later
transformed to biodiesel. Amyris (USA), LS9 (USA) Sapphire Energy
(USA), Solazyme (USA) and Terrebonne (Canada) study the way to scale
the process to industrial levels (Steen et al., 2010; Westfall and Gardner, 2011). The use of
GMO has had positive results in yield and efficiency. According to Tanimura
et al. (2015)
, the genetically modified yeast Scheffersomyces shehatae JCM
18690, can simultaneously hydrolyze and ferment starch with a
productivity of up to 0.92 g L-1 d-1 after ten days, comparable to the
highest yield reported in ethanol production from maize with S.
cerevisiae. Huerta et al. (2005) increased the theoretical yield of converting
glucose or xylose into ethanol up to 27 % by genetically improving
ethanologenic strains of E. coli KO11. One of the main objectives was to
optimize the expression of the gene PdcZm, which codes for pyruvate
decarboxylase enzymes, and to find or generate more active versions of
the same gene.

Future trends

The largest proportion of the costs of producing biofuels is associated

with raw materials (Neto et al., 2016). Thus, proposals for the industrial sector
and systems of technological innovation in biofuels is focused on
reducing production costs and on finding new raw materials that meet
economic and environmental requirements and do not affect human
food security.

One viable alternative for bioethanol production is process integration,

which helps to improve productivity and use of energy resources and
has an impact in reducing operation costs. One example is the
production of 1G and 2G ethanol from sugarcane and its bagasse, which
can reduce the final cost of ethanol enough to make it competitive with
fossil fuels (Neto et al., 2016). Technical analyses and economic studies should
be conducted on integrated processes. From sugarcane, a raw material
of high productivity, applicable results are obtained from integrated
processes at an industrial scale, but other less productive raw
materials, such as those with high starch content, should be analyzed in
technological integration processes. Moreover, using carbon dioxide
from the ethanol production plant (fermentation stage) in algae
production offers a new perspective for integration of technologies
1G/3G.

The concept of biorefineries is a way of integrating processes; it

broadens the spectrum of products of a production plant and,
therefore, market possibilities. But its level of complexity is advanced,
posing challenges in the integral use of raw materials and calling for
further study because of the number variables involved.

A recent proposal concerns synthetic ethanol production from organic

wastes such as paper, wood and cow manure. The residues are dried in
the same plant where ethanol is produced. They are stored for a short
period and introduced into the gasifier together with carbon to produce
carbon monoxide and hydrogen (“syngas”) as well as pure carbon.
“Syngas” is refined and subjected to a catalytic process before being
compressed for transfer to the reactor for ethanol synthesis. In this
process, collection of the raw material has a major impact on the cost
of the end product, and the carbon used to improve economic
efficiency increases costs. The cost of ethanol, with an internal return
rate of 10 %, was 0.433 USD L-1 with paper wastes, 0.51 USD L-1 with
wood waste and 1.45 USD L-1 with cow manure. Economic feasibility of
producing ethanol was greater with the mixture of wood waste and
carbon. Using carbon in the mixture may be an option for obtaining
energy with household generated organic waste (Gwank et al., 2018).

Petrochemical installations for producing and refining oil contribute CO 2

emissions as does combustion of the energy compounds (gasoline,
diesel, butane and natural gas). Combustion of oil derivatives produces
only emissions and no fixation which occurs with other gases (for
example, nitrogen), and there is no stage at which these emissions can
be decreased; thus, it causes environmental pollution. In contrast, CO 2
generated by ethanol combustion is less than that generated by fossil
fuels and can be fixed through photosynthesis and thus its release into
the atmosphere is lower, contributing less to the greenhouse effect
(Quintella et al., 2011).

Life cycle analysis (LCA) of biofuels helps to understand the impact of

the generated CO2 on the environment. An evaluation conducted by the
Centro de Tecnología Copersucar (CTC) showed that for each ton of
sugarcane, the net effect is fixation of 694.7 kg CO2, considering the
entire cycle, from sugarcane cultivation up to its final use as ethanol.
These results showed that emission of 206.8 kg CO2 per t of sugarcane
is avoided when ethanol is used instead of gasoline (Paula et al., 2010).

Diverse factors should be considered during the life cycle analysis of

ethanol. Sugarcane under given conditions can generate negative CO2
balances, but because of the variety of raw materials with which
ethanol can be obtained, each of them should be analyzed specifically.
For example, application of chemical fertilizers and irrigation water for
the crop that can be used to produce ethanol inevitably involves CO 2
emissions in the processes of production (Gelfand et al., 2011). Clearing and
converting land to crop fields damage the soil carbon store; this is
denominated carbon debt (the amount of CO2 released by land use
change). Production of precise reliable tools for determining and
quantifying each relevant contribution, anthropogenic or not, in the life
cycle of ethanol is a challenge.

CONCLUSIONS

Diversification of raw materials for ethanol production led to the

classification of first and second generation. First generation ethanol is
the use of materials rich in simple sugars (sucrose from sugarcane) and
starch (from maize). In the production of second-generation ethanol,
the aim is to take advantage of low-cost agricultural byproducts (maize
stover, wheat straw, etc.) that are rich in lignocellulosic compounds.
Third generation biofuels involve raw materials such as perennial
grasses, micro and macro algae, and cyanobacteria. In the case of
macro algae, their simple sugars and starch can be used to produce
ethanol. In the production of fourth generation ethanol, genetically
modified crops (such as sugarcane that is more efficient in capturing
CO2) and genetically modified microorganisms with greater efficiency in
converting substrate into product. Today, the search is for high-starch
raw materials, preferably non-food crops that are high-yielding and
low-cost, as promising alternatives for ethanol production at lower cost
than production from sugarcane and maize (food in Mexico and whose
use is prohibited in energy laws.

2.2 Characteristics of raw materials

2.3 Quantity of raw material required

3.Production process
3.1 Currently available Technologies

Manufacture of Ethanol • Ethanol maybeproduced by either synthetic

chemical method or by fermentation. • Fermentation was until about
1930 the main means of alcohol production. • In 1939, for example
75% of the ethanol produced in the US was by fermentation, in 1968
over 90% was madebysynthesis from catalytic hydration of ethylene.
• Dueto the increase in price of crude petroleum, the source of
ethylene used for alcohol production, attention has turned worldwide
to the production of alcohol by fermentation.

Microbial production of Ethanol: • Microbial production of ethanol

from the organic feed stocks and from plant substances such as
molasses is presently used for ethanol production. • Alcohol was
produced by fermentation in the early days but for many years by
chemical means through the catalytic hydration of ethylene. • In
modem era, attention has been paid to the production of ethanol for
chemical and fuel purposes by microbial fermentation. • Ethanol is
now-a-days produced by using sugar beet, potatoes, com, cassava,
and sugar cane (Fig. 20.6).

Both yeasts (Saccharomyces cerevisiae, S. uvarum S. carlsbergensis,

Candida brassicae, C. utilis, Kluyveromyces fragilis, K. lactis) and
bacteria (Zymomonas mobilis) have been employed for ethanol
production in industries. • The commercial production is carried out
with Saccharomyces cerevisiae. On the other hand, S. uvarum has also
largely been used. The Candida utilis is used for the fermentation of
waste sulphite liquor since it also ferments pentoses. • Recently,
experimentation with Schizosaccharomyces has shown promising
results. When whey from milk is used, strain of K. fragilis is
recommended for the production of ethanol. It is also found that
Fusarium, Bacillus and Pachysolen tannophilus (yeast) can transform
pentose sugars to ethanol. • Itisnoteworthy that the ethanol at high
concentration inhibits the yeast. Hence, the concentration of ethanol
reduces the yeast growth rate which affect the biosynthesis of
ethanol. • Thebacteria Zymomonas mobilis has a merit over yeast
that it has osmotic tolerance to higher sugar concentration. It is
relatively having high tolerance to ethanol and have more specific
growth rate.

1. Preparation of Medium: • Threetypes of substrates are used for

ethanol production: (a) Starch containing substrate (b) Juice from
sugarcane or molasses or sugar beet, (c) Waste products from wood
or processed wood. (d) Production of ethanol from whey is not viable.
• If yeast strains are to be used, the starch must be hydrolysed as
yeast does not contain amylases. After hydrolysis, it is supplemented
with celluloses of microbial origin so as to obtain reducing sugars.
About 1 ton of starch required 1 litre of amylases and 3.5 litre of
glucoamylases. Following steps are involved in conversion of starch
into ethanol (Fig. 20.7). • On the other hand, if molasses are used for
ethanol production, the bagasse can also give ethanol after
fermentation. • Several other non-conventional sources of energy
such as aquatic plant biomass, wood after hydrolysis with celluloses
gives ethanol. • Sulphite waste-liquor, a waste left after production of
paper, also contains hexose as well as pentose sugar. The former can
be microbially easily converted.

2. Fermentation: • Ethanol is produced by continuous fermentation.

Hence, large fermenters are used for continuous manufacturing of
ethanol. The process varies from one country to another. India, Brazil,
Germany, Denmark have their own technology for ethanol
production. • Thefermentation conditions are almost similar (pH 5,
temperature 35°C) but the cultures and culture conditions are
different. The fermentation is normally carried out for several days
but within 12h starts production. • After the fermentation is over, the
cells are separated to get biomass of yeast cells which are used as
single cell protein (SCP) for animal’s feed. The culture medium or
supernatant is processed for recovery of ethanol (Fig. 20.6). • Ethanol
is also produced by batch fermentation as no significant difference is
found both in batch and continuous fermentation. • Although as
stated earlier within 12h Saccharomyces cerevisiae starts producing
ethanol at the rate of 10% (v/v) with 10-20g cells dry weight/lit. The
reduction in fermentation time is accomplished use of cell recycling
continuously in fermentation.
3. Recovery: Ethanol can be recovered upto 95% by successive
distillations. To obtain 100%, it requires to form an azeotropic mixture
containing 5% water. Thus 5% water is removed from azeotropic
mixture of ethanol, water and benzene after distillation. In this
procedure, benzene water ethanol and then ethanol-benzene
azeotropic mixture are removed so that absolute alcohol is obtained.

General procedure for production of ethanol from

Sugarcane Regardless of whether the production is
done in a mass quantity or a backyard, the basic
steps for making ethanol are the same; 1. Procuring
the grain or plant 2. Converting this to sugar 3.
Fermentation 4. Distillation • On industrial scale,
ethanol is produced by the fermentation of molasses.
Molasses is the mother liquor left after the
crystallization of sugarcane juice. It is a dark colored
viscous liquid. Molasses contains about 60%
fermentable sugar. 1) Dilution of molasses Molasses is
first diluted with water in 1:5 (molasses: water) ratio
by volume Addition of • 2) Ammonium sulphate. If
nitrogen content of molasses is less, it is fortified
with ammonium sulphate to provide adequate
supply of nitrogen to yeast.

• 3) Addition of sulphuric acid Fortified solution of

molasses is then acidifies with small quantity of
sulphuric acid. Addition of acid favours the growth of
yeast but unfavours the growth of useless bacteria •
4) Fermentation The resulting solution is received in a
large tank and yeast is added to it at 35°C and kept
for 2 to 3 days. During this period, enzymes sucrose
and zymase which are present in yeast, convert sugar
into ethyl alcohol C12H22O11 + H2O à C6H12O6 +
C6H12O6 C6H12O6 à C2H5OH + 2CO2 5. Fractional
distillation Alcohol obtained by the fermentation is
called "wash" which is about 15% to 18% pure. By
using fractional distillation technique, it is converted
into 92% pure alcohol which is known as rectified
spirit or commercial alcohol. Production of ethanol
from various feed stocks involves the following steps.
I) Feed preparation 2) fermentation 3) distillation 4)
dehydration and 5) denaturing.

i ) Feed preparation The first step in making ethanol

is to prepare the feedstock to enter the fermentation
process. Cereal grains, such as corn, rye, rice, barley,
soybeans, wheat, and plants like sugar cane are the
major sources of feedstock's of fermentation. Some
producers use high starch plants such as potatoes.
Many different methods are used to prepare the
feedstock to enter the fermentation process. All of
the different processes ultimately produce a liquid
solution that contains fermentable sugars. These
solutions are clarified and heated to high temperature
for 20 to 30 minutes to reduce the bacterial levels
which can harm the performance of the process. After
this treatment the liquid mixture is removed and
subjected to fermentation process. If sugar cane is
used as a feedstock, the liquid mixture is said to be
sugarcane juice or molasses. ii) Fermentation of sugars
The liquid mixture obtained in the above process is
subjected to fermentation process by adding yeast
cells. Zymase, an enzyme from yeast, changes the
simple sugars into ethanol and carbon dioxide. The
enzymatic reaction carried over by the yeast in
fermentation produces mainly ethanol, CO2 and heat.
The fermentation reaction is actually very complex
and the resulting product is similar to beer or wine.
The impure culture of yeast Produces varying amounts
of other substances, including glycerin, methanol and
various organic acids. After fermentation, the liquid is
subjected to distillation to separate alcohol from water.

ii) Distillation Ethanol produced by fermentation

ranges in concentration from a few percent up to
about 14 percent balance is being water and other
components. The boiling point of ethanol (78.4'C) is
slightly lower than the boiling point of water
(100°C). Since the difference in the boiling point of
these materials is low they cannot be completely
separated by distillation. Instead, an azeotropic
mixture (i.e. a mixture of 96%ethanol and 4% water)
is obtained. Azeotropic mixture of alcohol cannot be
further concentrated by distillation. Distillation is used
to produce Rectified Spirit (RS, 94%v/v ethanol). iv)
Dehydration of Alcohol Pure alcohol can? be obtained
from distillation since it forms azeotrope with water
at 96% (vlv). Ethanol or absolute alcohol is produced
by dehydration of rectified spirit. Commercially
available technologies for dehydration of rectified
spirit are a) Azeotropic distillation and b) Molecular
Sieve Technology.

Industrial Ethanol Production

Commercial production of fuel ethanol in the U.S. involves

breaking down the starch present in corn into simple
sugars (glucose), feeding these sugars to yeast
(fermentation), and then recovering the main prod-uct
(ethanol) and byproducts (e.g., animal feed). Two major
industrial methods for producing fuel ethanol are used in
the U.S.: wet milling and dry grind. Dry-grind ethanol
production represents the majority of ethanol processing
in the U.S. (> 70% of production), and all newly
constructed ethanol plants employ some variation on the
basic dry-grind process because such plants can be built
at a smaller scale for a smaller investment.Wet
MillingWet milling is used to produce many products
besides fuel ethanol. Large-scale, capital-intensive, corn-
processing wet mills produce such varied products as
high fructose corn syrup (HFCS), biodegradable plastics,
food additives such as citric acid and xanthan gum, corn
oil (cooking oil), and livestock feed.

Wet milling is called “wet” because the first step in the

process involves soaking the grain in water (steep-ing) to
soften the grain and make it easier to separate
(fractionate) the various components of the corn kernel.
Fractionation, which separates the starch, fiber, and
germ, allows these various components to be processed
separately to make a variety of products. The major
byproducts of wet-mill ethanol production are two animal
feed products, corn gluten meal (high protein, 40%) and
corn gluten feed (low protein, 28%), and corn germ,
which may be further processed into corn oil.Dry GrindIn
the dry-grind ethanol process, the whole grain is
processed, and the residual components are separated at
the end of the process. There are five major steps in the
dry-grind method of ethanol production. dry-Grind
ethanol Processing steps1. Milling2. Liquefaction3.
Saccharification4. Fermentation5. Distillation and
recoveryMillingMilling involves processing corn through a
hammer mill (with screens between 3.2 to 4.0 mm) to
produce

a corn flour (Rausch et al., 2005). This whole corn f lour is

slurried with water, and heat-stable enzyme (a-amylase)
is added. Liquefaction T his slurry is cooked, also known
as “liquefaction.” Liquefaction is accomplished using jet-
cookers that inject steam into the corn flour slurry to
cook it at temperatures above 100°C (212°F). The heat
and mechanical shear of the cooking process break apart
the starch granules present in the kernel endosperm, and
the enzymes break down the starch polymer into small
fragments. The cooked corn mash is then allowed to cool
to 80-90°C (175-195°F), additional enzyme (a-amylase)
is added, and the slurry is allowed to continue liquefying
for at least 30 minutes. Saccharification After
liquefaction, the slurry, now called “corn mash,” is cooled
to approximately 30°C (86°F), and a second enzyme
(glucoamylase) is added. Glucoamylase completes the
breakdown of the starch into simple sugar (glucose). This
step, called “saccharification,” often occurs while the
mash is filling the fermentor in preparation for the next
step (fermentation) and continues throughout the next
step. Fermentation In the fermentation step, yeast grown
in seed tanks are added to the corn mash to begin the
process of converting the simple sugars to ethanol. The
other components of the corn kernel (protein, oil, etc.)
remain largely unchanged during the fermentation
process. In most dry-grind ethanol plants, the
fermentation process occurs in batches. A fermentation
tank is filled, and the batch ferments completely before
the tank is drained and refilled with a new batch. T he up-
stream processes (grinding, liquefaction, and
saccharification) and downstream processes (distillation
and recovery) occur continuously (grain is continuously
processed through the equipment). Thus, dry-grind
facilities of this design usually have three fermentors
(tanks for fermentation) where, at any given time, one is
filling, one is fermenting (usually for 48 hours), and one is
emptying and resetting for the next batch. Carbon dioxide
is also produced during fermentation. Usually, the carbon
dioxide is not recovered and

is released from the fermenters to the atmosphere. If

recovered, this carbon dioxide can be compressed and
sold for carbonation of soft drinks or frozen into dry ice
for cold product storage and transportation. After the
fermentation is complete, the fermented corn mash (now
called “beer”) is emptied from the fermentor into a beer
well. The beer well stores the fermented beer between
batches and supplies a continuous stream of material to
the ethanol recovery steps, including distillation.
Distillation and Recovery After fermentation, the liquid
portion of the slurry has 8-12% ethanol by weight.
Because ethanol boils at a lower temperature than water
does, the ethanol can be separated by a process called
“distillation.” Conventional distillation/rectification
systems can produce ethanol at 92-95% purity. The
residual water is then removed using molecular sieves
that selectively adsorb the water from an ethanol/water
vapor mixture, resulting in nearly pure ethanol (>99%). T
he residual water and corn solids that remain after the
distillation process are called “stillage.” This whole
stillage is then centrifuged to separate the liquid (thin
stillage) from the solid fragments of the kernel (wet cake
or distillers’ grains). Some of the thin stillage (backset) is
recycled to the beginning of the dry-grind process to
conserve the water used by the facility. T he remaining
thin stillage passes through evaporators to remove a
significant portion of the water to produce thickened
syrup. Usually, the syrup is blended with the distillers’
grains and dried to produce an animal feed called
“distillers’ dried grains with solubles” (DDGS). When
markets for the feed product are close to the plant, the
byproduct may be sold without drying as distillers’ grains
or wet distillers’ grains. Energy Use in Ethanol Production
It is true that the laws of physics dictate that energy will
be lost in converting one form of energy to another. Thus,
ethanol does have less energy than the corn used to
produce it. However, this is also true for converting crude
oil to gasoline and coal to electricity. T he important
questions about ethanol production are “is ethanol truly a
renewable fuel?” and “how much fossil fuel is used?”

Yes; ethanol is a renewable fuel. The energy used to

produce ethanol includes fuel for tractors, combines, and
transportation of the grain to the ethanol plant, as well as
the energy in processing the corn to ethanol. However,
the largest portion of the total energy present in corn is
solar energy captured by the corn plant and stored in the
grain as starch. When these amounts are totaled, the
energy in the ethanol exceeds the fossil fuel energy used
to grow and process the corn by 20 to 40% (Farrell et al.,
2006). Most of the energy for processing corn to ethanol
is spent on the distillation and DDGS drying steps of the
process. When wet distillers’ grain can be fed to livestock
close to the ethanol plant, the savings in natural gas for
drying can be as high as 20% of the total energy cost for
processing corn to ethanol. Conclusions Modern dry-grind
ethanol plants can convert corn grain into ethanol (2.7-
2.8 gallons per bushel) and DDGS (17 pounds per bushel).
This rather energyefficient process produces a renewable
liquid fuel that has significant impacts on the agricultural
economy and energy use in the U.S. Increasing ethanol
production presents many opportunities and challenges
for U.S. agriculture as demands on corn production for
feed, fuel, and export markets increase. Additionally,
advances in biotechnology and engineering are opening
possibilities for new raw materials, such as switch grass
and corn stover, to be used for even greater fuel ethanol
production into the future.

4. Byproducts

Ethanol fermentation produces unharvested byproducts

such as heat, carbon dioxide, food for livestock, water,
methanol, fuels, fertilizer and alcohols.[16] The cereal
unfermented solid residues from the fermentation
process, which can be used as livestock feed or in the
production of biogas, are referred to as Distillers grains
and sold as WDG, Wet Distiller's grains, and DDGS, Dried
Distiller's Grains with Solubles,