ETHANOL

commonly called alcohol, ethyl alcohol, and drinking alcohol

the principal type of alcohol found in alcoholic beverages,
produced by the fermentation of sugars by yeasts.
It is a neurotoxinc, psychoactive drug, and one of the
oldest recreational drugs. It can cause alcohol
intoxication when consumed in sufficient quantity.
Ethanol is a volatile, flammable, colorless liquid with a slight
chemical odor.
Empirical formula is C2H5OH

CHEMICAL STRUCTURE

HISTORY OF ETHANOL
The fermentation of sugar into ethanol is one of the earliest organic
reactions that man learned to carry out and the history of man-made
ethanol is very long.
Dried ethanol residue have been found on 9 000 year old pottery in China
which indicates that Neolithic people in this part of the world may have
consumed alcoholic beverages.
Distillation was well known by the early Greeks and Arabs. Greek
alchemists working in Alexandria during the first century A.D. carried out
distillation.
Fractional distillation was invented by Tadeo Alderotti in the 13th century.
The year 1796 is significant for ethanol history because this is when
Johann Tobias Lowitz obtained pure ethanol by filtering distilled ethanol
through activated charcoal.
In mid 1800s, ethanol became one of the first structural formulas to be
determined The scientist behind the description was Scottish chemist
Archibald Scott

HOW TO MAKE ETHANOL ?

Ethanol can be produced in different ways:
through fermentation - biological process
through ethylene hydration - a petrochemical
process
Wet milling process
Dry milling processes

PRODUCTION OF ETHANOL BY FERMENTATION

Fermentation is the oldest way for humans to produce

ethanol, and this is the traditional way of making
alcoholic beverages. It is also the process used for the
vast majority of ethanol fuels on the market.
When certain species of yeast metabolize sugar, the
end result is ethanol and carbon dioxide. One
example of such a species is Saccharomyces
cerevisiae, which has been used by brewers since
ancient times.
In Greek Saccharo means sugar and myces means
fungus

 This is the chemical formula for turning sugar into ethanol and
carbon dioxide:
C6 H12 O6
2 CH3CH2OH + 2 CO2
C6 H12 O6 is simple sugar, also known as
dextrose or D-glucose.
CH3CH2OH is ethanol
CO2 is carbon dioxide
Most types of yeast will stop reproducing when the alcohol
content reaches 15% ethanol by volume, or even earlier, putting
a natural limit on the alcohol concentration achieved through
fermentation

BIOCHEMICAL PROCESS

MICROORGANISMS USED
Bacteria

Zymomonas mobilis
Clostridium acetobutylicum
Klebseilla pnemonia
Candida brassica

Fungi
Saccharomyces cerevisiae
Schizosaccharomyces

BIOETHANOL PRODUCTION
Bioethanol can be produced in three ways:
First generation bioethanol

SUGAR
STARCH

SUGAR

ETHANOL
ETHANOL

Second generation bioethanol

CELLULOSE OR HEMICELLULOSE

Third generation bioethanol

ALGAE

SUGAR

ETHANOL

ETHANOL

CROPS USED IN FIRST GENERATION

BIOETHANOL PRODUCTION

INDIA
BRAZIL
USA
EUROPE

:
:
:
:

SUGARCANE
SUGARCANE
CORN
WHEAT AND BARLEY

CHOICE OF RAW MATERIAL DEPENDS

UPON

Ease of processing of various plant materials

Prevailing conditions of climate
Landscape and soil composition
Sugar content

SECOND GENERATION BIOETHANOL

PRODUCTION
Second generation biofuels are made from lignocellulosic biomass or
woody crops, agricultural residues or waste, which makes it harder to
extract the required fuel.
Second generation biofuel technologies have been developed
because first generation biofuels manufacture has important limitations.
The goal of second generation biofuel processes is to extend the amount of
biofuel that can be produced sustainably by using biomass consisting of the
residual non-food parts
of
current
crops,
such
as stems, leaves and husks that are left behind once the food crop has been
extracted, as well as other crops that are not used for food purposes (nonfood crops) which contains lignin, cellulose and hemicellulose.
Lignocellulosic biofuels reduces greenhouse gas
emissions with 60-90% when compared with fossil
petroleum (Brjesson.P. et al. 2013)

Bioethanol from cellulose

2nd generation of bioethanol
Researchers are exploring ways to make ethanol from other feedstocks or
plant materials.
Cellulosic ethanol production starts with the biomass or plant materials and
breaks down the cell wall to release the starch or sugars in the plants leaves
and stems. These simpler compounds are then fermented into ethanol.
The combination of lignin, hemicellulose and cellulose in plant material is
very resistant to breakdown into the molecular sugar components. Two main
pathways to converting lignocellulose to biofuels are:
Biological specialized enzymes or microbes break pre-treated biomassbased cellulose into sugars, which are then fermented into alcohols.
Thermochemical biomass is converted by pyrolysis, gasification, or
liquefaction into gaseous and liquid chemical "building blocks. It is
recombined through catalytic processes into a variety of fuels and chemicals.

THIRD GENERATION BIOETHANOL

PRODUCTION

Major feedstock is ALGAE

Advantages
Dont occupy agricultural lands
Do not need any fertilizer for cultivation
Significant carbohydrate content, higher ethanol yields are obtained
Algae can grow in every season and everywhere such as salty waters, fresh
waters, lakes, deserts and marginal fields etc

Two types of algae

Microalgae - high amount of lipid, protein and carbohydrate mainly starch
Macroalgae - carrageenan, laminaran, mannitol, alginate
Chlamydomonas reinhardtii has highest amount of acrbohydrate content

CONDITIONS FOR FERMENTATION

Carbon source- Pure sugar or molasses(10-18% , a
concentration higher than this is detrimental to the yeast
strain)
Nitrogen Source- Mostly available in form of
Ammonium Sulphate
Growth Factor- can be provided in form of molasses.
pH- 4.8 5
Temperature- 32C-45C
Time- depend on strain, usually between 30 to 70 hrs(23 days)

PRODUCTION PROCESS
PRETREATMENT
Most of the raw material require some degree of pretreatment .
In general sugary material require mild or no pretreatment while cellulosic material require extensive
pretreatment.
This is because cellulosic material have to be
subjected to acidic or enzyme hydrolysis to release
monosaccharide units that are required for alcohol
fermentation.

 Production process of alcohol is carried out in

3 stages
1. Inoculum
2. Proper fermentation
3. Recovery

PREPARATION OF INOCULAM
After selection of desired organism and its in
pure form the Inoculam is prepared under
aseptic condition.
For this condition organism is fist culture in
flask to increase the size of the inoculam
which increases the size of the inoculam which
can then be used for inoculation.

PROPER FERMENTATION
Previously Batch fermentation was used but now
Continuous fermentation is used as it increases
the alcohol production by 10-12 fold as compare
to batch fermentation.
Large fermentor of size 1,25000 gallon are used .
Initially aeration is required for good growth of
micro organism .Later anaerobic conditions are
created by withdrawal of oxygen .
It take 2-3 days for the fermentation to complete.

A STUDY
Fermentation of sugar by Saccaharomyces cervisiae for
production of ethanol in a batch experiment .
When the temperature was increased to 45C, the system still
showed high cell growth and ethanol production rates, while it
was inhibited at 50C. The maximum specific growth rate and
the maximum specific ethanol production rate were observed
between 30-45C.
pH 4.0-5.0 was the optimal range for the ethanol production
process. The highest specific ethanol production rate for all the
batch experiments was achieved at pH5.0. Formation of acetic
acid was increased when the pH was below 4.0, while butyric
acid was produced when the pH was higher than 5.0

BATCH FERMENTATION
In Batch fermentation bacteria are inoculated
into the bioreactor .
Then under optimal condition(temp, pH etc) the
bacteria go through all the growth phases(LAG,
exponential and stationary).
It may be necessary to add acid or alkali to
maintain pH and antifoaming agents to
minimize foam under optimal condition for
growth.

PHASES

Lag phase
Acceleration phase
Logarithmic (log) phase
Deceleration phase
Stationary phase
Death phase

LAG PHASE
initial period just after the inoculation
During lag phase micro organisms adapt to the new
environment (available nutrient ,pH etc)
No increase in cell number, although cellular weight
may slightly increase.
ACCELERATION PHASE
Brief transient period during which cell start growing
slowly .
Connect lag phase to log phase.

LOG PHASE
Most active growth of micro- organism and multiplication occur
during log phase.
Number of cells and rate of population increases doubles with each
consecutive time period.
Growth rate of microbes in log phase is dependent on
substrate(nutrient supply)
STATTIONARY PHASE
Depletion of nutrients and accumulation of metabolic end product .
Microbial growth may either completely slow down or completely
stop
Biomass may remain constant .
The number of cells produced is limited by growth factor and as a
result the rate of cell growth matches the rate of cell detah.

DEATH PHASE
Cells die at an exponential rate

 At last fermentation is stopped and product is

collected.
Then after cleaning and sterilization of
fermentor is ready for another batch

ADVANTAGES
ADVANTAGES
VERSATILE- can be used for different
reaction everyday
SAFE- can be properly sterilized. Little risk of
infection or strain mutation. Complete
conversion of substrate is possible.

DISADVANTAGS
High labor cost: Skilled labor is required.
High proportion of Down time between
batches
Product Variability The quality and quantity
(to some extent ) may vary from one batch to
other batch.
Safety Problem- when cleaning, filling and
emptying.

CONTINIOUS FERMENTATION
In, continuous fermentation fresh medium flows into
the fermentor continuously, and a part of the medium
is withdrawn from the fermentor at the same flow rate
of the inlet flow.(so that working volume remain
constant)
Chemostat bioreactor are frequently used in industrial
manufacturing of ethanol.
BY changing the rate at which the medium is added to
the bioreactor the specific growth rate of the microorganism can be easily controlled

ADVANTAGES
Low labor cost
Growth rate is higher as nutrients are
continuously added to the reactor.
More efficient as the fermentor operates
continuously.(No down time )
DISADVANTAGE
If contamination occur huge volume of
product may be lost.

Power Alcohol
Ethyl alcohol which is used to generate power; for example as an
additive to motor fuels to act as fuel for internal combustion engine is
called power alcohol. It has generally 80% petrol and 20% alcohol.
There have been decades of motor fuel application experience in the
United States and other countries with ethanol. Ethanol has been
blended with fuels to produce more efficiency.
The practice of blending ethanol started in India in 2001. Government
of India mandated blending of 5% ethanol with petrol.
Rather than using chemically produced alcohol, nowadays ethanol
prepared by biomass is being used for this blending. This is
considered more environment friendly and hence are called biofuels.
These biofuels have overtaken the traditional fossil fuels over the past
few years all over the world.

BIOFUELS
Biofuels are those liquid or gaseous fuels produced from biomass
resources and used in place of, or in addition to, diesel, petrol or
other fossil fuels for transport, stationary, portable and other
applications.
Three broad categories of bio-fuels are identified in India:
- bio-ethanol: ethanol produced from biomass such as sugar
containing materials, like sugar cane, sugar beet, sweet sorghum,
etc.; starch containing materials such as corn, cassava, algae etc.;
and, cellulosic materials such as bagasse, wood waste, agricultural
and forestry residues etc.;
-biodiesel: a methyl or ethyl ester of fatty acids produced from
vegetable oils, both edible and non-edible, or animal fat of diesel
quality; and
-other biofuels: biomethanol, bio CNG, biosynthetic fuels etc.

PRODUCTION OF BIOETHANOL
Wet Milling:
Wet milling is used to produce many products besides fuel ethanol. Wet
milling is called wet because the first step in the process involves
soaking the grain in water (steeping) to soften the grain and make it
easier to separate (fractionate) the various components
Dry Grind:
In 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:
1. Milling
2. Liquefaction
3. Saccharification
4. Fermentation
5. Distillation and recovery

PRODUCTION OF BIOETHANOL

BIOETHANOL PRODUCTION USING CORN

First generation ethanol production
1. Milling: involves processing the corn through a hammer mill .This whole

corn flour is slurried with water, and heat-stable enzyme (a-amylase) is added.
2. Liquefaction:
- accomplished using jet-cookers that inject steam into the corn flour slurry
to cook it at temperatures above 100C (212F).
- The heat and mechanical shear of the cooking process break apart the
starch, and the enzymes break down the starch polymer into small
fragments.
- The cooked corn mash is then allowed to cool to 80-90C (175-195F),
additional enzyme (a-amylase) is added, and the slurry is allowed to
continue liquefying for at least 30 minutes.
3. Saccharification :
-The slurry is cooled to approximately 30C (86F), and a second enzyme
glucoamylase is added. Glucoamylase completes the breakdown of the starch
into simple sugar.

4. Fermentation:
Yeast grown in seed tanks are added to the corn mash to begin the
process of converting the simple sugars to ethanol.
5. Distillation and Recovery :
After fermentation, the liquid portion of the slurry has 8-12%
ethanol by weight. Conventional distillation/rectification systems
can produce ethanol at 92-95% purity.
The residual water and corn solids that remain after the distillation
called stillage. is then centrifuged to separate the liquid (thin
stillage) from the solid fragments (wet cake or distillers grains).
The thin stillage is passed 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).

BIOETHANOL PRODUCTION USING SUGARCANE

First generation ethanol production

Cutting , washing and drying of sugarcane.

Buffer was used to dilute the enzyme alpha-amylase ( phosphate buffer) and
glucoamylase (sodium acetate buffer)
Liquefaction of sugarcane: Sugarcane was weighed and NaOH was added
to maintain pH at 4.5. Alpha amylase enzyme was added to it and heated
until 500C. Alpha- amylases will breakdown the cellulose into smaller size
called dextrin.
Saccharifaction of sugarcane bagasse- The mixture was cooled down to
400C. Glucoamylase was added to the mixture which hydrolyzed the dextrin
to fermentable glucose. The mixture was cooled to 32 0C and
Saccharomyces cerevisiea (baker yeast) was added.
Fermentation of sugarcane - Saccharomyces cerevisiea (baker yeast) was
used to ferment the simple sugar to ethanol and carbon dioxide.
Distillation: The bioethanol was distilled using rotary evaporator. The
sample was heated at 800C to get the bioethanol.

WHY IS BIOETHANOL GOOD

Availability - It is renewable. Unlike fossil fuels, biofuels can be easily
produced from raw agricultural materials.
Cleaner fuels - which means they produce fewer emissions on burning.
Reduce Greenhouse Gases: Fossil fuels, when burnt, produce large
amount of greenhouse gases i.e. carbon dioxide in the atmosphere
leading to global warming. Studies suggests that biofuels reduces
greenhouse gases up to 65 percent.
They release lower levels of carbon dioxide and other emissions when
burnt. Although the production of biofuels creates carbon dioxide as a
byproduct, it is frequently used to grow the plants that will be converted
into the fuel. This allows it to become something close to a self
sustaining system.
If more people start shifting towards biofuels, a country can reduce its
dependence on fossil fuels and also imported fuels from other country
can be reduced leading to economic development of our country.

IS BIOFUEL HARMFUL?
High Cost of Production: Even with all the benefits associated
with biofuels, they are quite expensive to produce in the current
market.
Monoculture: Growing same crop every year may deprive the
soil of nutrients that are put back into the soil through crop
rotation.
Industrial Pollution: The process with which they are produced
is largely dependent on lots of water and oil. Large scale
industries meant for churning out biofuel are known to emit
large amounts of emissions and cause small scale water
pollution as well.

Distillation and Ethanol

Recovery