A MINOR PROJECT REPORT SUBMITTED IN THE PARTIAL

FULFILLMENT OF
REQUIREMENT FOR THE AWARD
OF DEGREE OF
BACHELOR OF TECHNOLOGY
IN
CHEMICAL ENGINEERING
SUBMITTED TO
B.I.T. SINDRI
(DHANBAD, JHARKHAND)

SUBMITTED BY:
KARAN SOREN (1504033)
KOKIL ORAON (1504034)
KRITIKA KIRAN (1504035)
LUCKY ARON SOREN(1504036)
MAMTA MARKEY(1504037)
MANISHA SOREN (1504038)
MANJEET ORAON (1504039)
MANOJ TUBID (1504040)

UNDER THE GUIDANCE OF

PROF: AJAY AORAON

DEPARTMENT OF CHEMICAL ENGINEERING
 APPROVAL CERTIFICATE

This Minor Project work entitled “ Bio-ethanol from agricultural

waste” being submitted by …………………………, Roll
No:………………. has been examined by our team members and
is hereby approved for the award of degree of Bachelor of
Engineering in Chemical Engineering which has been submitted
to BIRSA INSTITUTE OF TECHNOLOGY, SINDRI

Internal Examiner
External Examiner
Date:
Date:
 ACKNOWLEDGEMENT

I wish to express my profound gratitude and indebtedness to

Prof. Ajay Oraon, Professor, Department of Chemical
Engineering, Birsa Institute of Technology, Sindri, for introducing
the present topic and for his inspiring guidance, constructive
criticism and valuable suggestions throughout this project work.
I would also express my gratitude to all my friends and professors
who have partially extended all sorts of help for accomplishing
this undertaking.

Department of Chemical Engineering

B.I.T. Sindri, Dhanbad
 1.INTRODUCTION
Due to increasing population the demand of energy is increasing throughout the world.
Primary source of energy is the fossil fueland non-renewable sources of fuel asnatural gas
oil and coal.Overconsumption of fossil fuel has created more pollution due to
GHGs.According to world energy council, petroleum natural gas and coal which are the
good source of energy collectively contribute to the nearly 82% of the global energy needs
and 1/5th of the CO2 is due to the 100% of the petroleum based fossil fuel.To reduce
dependency on resources we generally used alternate method for production of energy like
wind water biomass and geothermal heat form.

In future chemical industry may depend on biomass as alternative source. Biomass from
cellosic agriculture waste is the potential promising natural renewable inexpensive cost
effective and sustainable sources used for considerable and commercial production of bio
energy as bio ethanol.The renewable fuels such as bio diesel bio hydrogen derived from
sugarcane corn switchgrass algae etc. can be used as petroleum based in future.

1.1:BIO ENERGY
Bemdes Et Al estimated that potential global bio energy supply range from less than 100 to
over 400 Ej/year for 2050.Bio ethanol is a gasoline alternate while biodiesel is a diesel
alternate tto reduce GHGs emission when blended as an additive.Bio ethanol is produced
60% from sugarcane and 40% from other crops.Biodiesel produced from inedible vegetable
oils , waste oil and grease and was estimated that in 2017 about 60 billion lt. bio fuel
produced globally.It has been accounted that bio ethanol could sink about 90% CO2 and 60-
80% SO2 when blended with 90% gasoline.

Biomass is fourth biggest source of energy after petroleum and natural gas.It is broadly
used as liquid bio fuel gas for motor vehicles.Its cost is higher than fossil fuels.Bio ethanol
production in 2008 was 66.7 billion lt ,in 2013 it was grown upto 88.69 billion and in 2014 it
was 90.38 billion litres.

America and Brazil are major producers of bioethanol contributing towards 56.72% and
26.72% of production respectively.
 TABLE 1.1: BIOETHANOL PRODUCTION FROM DIFFERENT OUNTRIES FROM YEAR 2004
TO 2014

COUNTRY 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013 2014

ALLGERIA - - - - - - - - - - -
EGYPT - - - - - - - - - - -
SUBSAHAR - - - - - - - - - - -
AN AFRICA
REPUBLIC 15 15 15 15.5 16 15.42 15.74 15.9 16.07 16.18 16.26
OF SOUTH
AFRICA
ARGENTIN 174 157 205 225 315 416 440 454.7 468.9 483.9 497.6
A
BRAZIL 15207 15807 17931 22445 27674 25804 28960 31391 34298 37396 40625
BANGLADE - - - - - - - - - - -
SH
China 3673 3438 3509 3679 3964 4109 4368 4648 4823 4961. 5121.
9 1
India 1178.2 1120.4 1663.5 2081.9 2084.5 1680. 1703.5 2429. 2481. 2532 2574
3 8 5 7
Indonesia 163 177 176 196 208 240 424 440 461 485 509
Iran - - - - - - - - - - -
Malaysia 79.28 62.28 63.28 64.30 64 66.25 66.48 66.84 67.03 67.48 67.98
Pakistan - - - - - - - - -- - -
Saudi Arabia - - - - - - - - - - -
EU 27 2576 2940 3701 3887 5021 5761. 6465 7538 9154 10795 11773
5
RUSSIA - - - -- - - - - - -
Ukraine - - - - - - - - - - -
Canada 396 405 544 839 1083 1130 1572 1703 1713 1729 1721
United state 12596.4 15332 20171 28929 35190 40543 46024 49113 51321 54057 57199
5
Australia - 27 62 100 155 238 383 386 389 392 394
Mexico 35 58 49 61 61 65 70 75 76.95 78.83 80.71
Japan - 113 113 110 110.20 100.2 100.20 130 130 130 130
0
Turkey 19.02 46.58 50.76 44.34 54.4 64.34 64.99 65.18 65.49 65.76 65.86
OECD 15627 18876 24641 33926 40621 48194 54616 58946 62786 67183 71300
Non OECD 21084 21449 24562 29880 95560 33942 37964 41750 45199 48806 52544
DEVELOPE - 18818 24592 33865 41560 47774 54545 58871 62709 67104 71219
D
Developing - - - - -- - - - - - -
Least - - - - - - - - - - -
developing
It has been reported that 442 billion lt of bio ethanol can be produced from lignocellulosic
biomass and that total crop residues and wasted crops can produce 491 billion lts of bio ethanol
per year of separation of free cellulose from agro residue.Enzyme hydrolysis can be done by
microbes or fungi where microbes include- clostridium ,cellulomonas, thermonopora, bacillus
etc. and fungi include trichroderma, penicillium, fusarium etc.

Fermentation is required for conversion of glucose to ethanol by microbes.eg

sacharomycescerevisiae, Escherichia coli, zymomonasaetc.It is environmental friendly gas and
acceptable technology also reduces the green house gas emission.

TABLE 1.2: ETHANOL IS GIVEN IN MILLION GALLON

4500 THE ETHANOL IS GIVEN IN MILLION GALLON.

4000
Global standpoints of bio ethanol production from cellulosic materials.
3500
 INTERNATIONAL SCENARIO
3000
The first oil crisis came during world war 2 in 1973 due to soaring demand of
2500 petroleum and scaling up burning of transportation biofuel more.
2000 According to world watch institute the world‟s oil consumption
TOPMOSTincreased
ETHANOL from
1500 2004-2005 demand increased by 5.3% typically in China, US PRODUCING
,CanadaCOUNTRIES
and UK.
1000 US independently was the world‟s biggest polluter in 2005 and consumed about
500
140 billion gallons of transportation fuel and 398 million metric tons of carbon
emitted into the atmosphere by the gas guzzling vehicles.
0
Brazil US China India France RussiaSouth Korea UK Saudi Arabia
 1.2: INTERNATIONAL SCENARIO
TABLE 1.3: WORLDS TOTAL PRODUCTION OF FUEL ETHANOL FROM YEAR 2004 TO
2013

Country Sugar and 200 200 200 200 200 200 201 201 201 201
starchy 4 5 6 7 8 9 0 1 2 3
crops
US Corn/maize 13 15 18.3 24.6 34 41 49.5 54.2 50.4 50.3
BRAZIL Sugarcane 15 15 17.5 1 27 26 27.6 21 21.6 25.5
GERMANY Wheat 0.02 0.2 0.5 - 0.5 0.8 1.5 0.8 0.8 0.8
FRANCE Sugarbeet 0.1 0.15 - - 1.2 0.9 1.1 1.1 1.0 1.0
,wheat
CHINA Corn 2 1 1 1.8 1.9 2.1 2.1 2.1 2.1 2.0
sugarcane
maize
Argentina Sugarcane - - - 0.02 - - 0.1 0.2 0.2 0.5
ITALY Cereals - - 0.13 - 0.13 0.1 0.1 0.0 - -
SPAIN Barley 0.2 0.3 0.4 - 0.4 0.4 0.6 0.5 0.4 0.4
wheat
India Sugarcane - 0.3 0.3 0.2 0.3 0.2 - - 0.5 -
wheat
CANADA Wheat/cerea 0.2 0.2 0.2 0.8 0.9 1.1 1.4 1.8 1.8 1.8
l
CZECH Sugar beet - 0.15 0.0 - - - - - - 0.2
REPUBLIC
COLOMBIA Sugarcane - 0.2 0.2 0.3 0.3 0.3 0.4 0.3 0.4 0.4
SWEDEN Wheat - 0.2 0.14 - 0.14 - - - - -
MALAYSIA - - - - - - - - - - -
UK - - - - - - 0.2 0.3 - - -
DENMARK Wheat - 0.1 - - - - - - - -
AUSTRIA Wheat - 0.1 - - - 0.1 - - 0.2 -
SLOVAKIA Corn - 0.1 - - - - - - - -
THAILAND Sugarcane, 0.2 - - 0.3 0.3 0.4 0.4 0.5 0.7. 1.0
cassava
AUSTRALI Sugarcane 0.07 - - 0.1 - - - - - 0.3
A
Belgium Wheat - - - - - 0.2 0.3 0.4 0.4 0.4
EU Various - - - 2.16 - - 4.5 4.3 4.2 4.5
cereal and
sugar beet
Poland Rye - 0.05 0.12 - 0.12 - 0.2 - - 0.2
World total 31 33 39 49.6 67 76 86 86.1 83.1 87.2
TABLE 1.5: POTENTIAL BIO ETHANOL PRODUCTION FROM FOOD CROP BY CONTINENT.

CONTINENTS Corn Barley Oat Rice Wheat Sorghum Sugarcane Total

Africa 2.17 0.12 0.002 0.71 0.55 1.55 0.23 5.33
Asia 6.82 0.83 0.04 14.4 6.78 0.37 0.82 30.1
Europe 1.09 1.35 0.30 0.02 2.70 0.003 - 5.45
North America 0.21 0.005 0.01 0.63 0.02 - - 0.87
Central 1.21 0.01 0.0004 0.05 0.16 0.09 0.18 1.70
America
Oceania 0.01 0.13 0.001 0.02 0.54 0.0004 0.0001 0.70
South 2.87 0.03 0.03 0.93 0.60 0.12 0.37 4.95
America
1.3:NATIONAL SCENARIO
During 2007 ,156 million tons of crude oil was consumed of which 77% was imported.It was
predicted that this will rise about 6 million barrels per day by 2030, formulated India as the third
largest importer of oil.Currently India produces 1.3 billion litres of ethanol from cane molasses
against an installed capacity of 3.2 billion lts.

Now India is adopting a new policy of production of ethanol from cellulosic biomaterials which is
an E20 by 2017.It is also been reported that biofuel production in India using sugarcane and
wheat are the major feedstock.Also it is estimated that about 199.1 million metric tons of
sugarcane is cultivated in excess of huge area in India hence, it generates surplus amount of
residue.It is stated that ethanol can also be produced by an agro waste ie banana peels which is
rich in carbohydrates crude protein and reducing sugars and owing to affordable and renewable
low cost material makes it potential feedstock for ethanol production.

Cellulosic Material as agro residues for bio ethanol production.

600-900 million tons rice straw is produced globally

The percentage distribution of cellulose, hemicelluloses and lignin on dry weight basis of various
agro residues have been depicted in table 1.7.

Sources Cellulose% Hemicelluloses% Lignin%

Corn stover 31(mf wt%) 43(mf wt%) 13(mf wt%)
Wheat straw 32.4(mf wt%) 41.8(mf wt%) 41.8(mf wt%)
Cereal straws 35-40% 26% 15-20%
Poplar aspen 42.3(mf wt%) 31(mf wt%) 16.2(mf wt%)
Rice straw 32.6 27.3 18.4
Baggase 65(total - -
carbohydrate)
Seaweed 20.35(alpha) 25.73 0-15
Paper 85-99 0 0
Cotton seed 80-85 5-20 0
hairs
Oil palm frond 49.8(alpha) 83.5(Holocellulose) 20.5
Coconut 44.2(alpha) 56.3(Holocellulose) 32.8
Pineapple leaf 73.4(alpha) 80.5(Holocellulose) 10.5
Banana stem 63.9(alpha) 65.2(Holocellulose) 18.6
Softwood 40-50 25-30 25-35
Hardwood 40-50 25-35 20-25
Big blustem 29-37 21-25 17-24
Switchgrass 31-35 24-28 17-23
Jatropha waste 56.31 17.47 23.91

A perennial gas namely elephant grass can be used to produce ethanol.

According to demirbas stated that the second and third generation bio fuels are also called
advanced bio fuels During 1st generation of bio fuel sugar, starch or vegetable oil were used to
produce biofuels such as ethanol, bio diesel and bio gas .

Second generation used non-food crops such as wheat straw,corn,wood,solid waste,energy

crop.While third generation biofuels are produced by using algae but the main problem with the
algae was that it makes merely 0.1% of the mass rest of 99.9% water.

The fourth generation involves using vegetable oil for production of biofuel.

Ethanol production in India (billion

litres/year)
0.35

0.3

0.25

0.2
Ethanol production in India
0.15
(billion litres/year)
0.1

0.05

0
2004 2005 2006 2007 2008 2009

2.RAW MATERIALS
2.1: BAGASSE
Bagasse is the dry pulpy fibrous residue that remains after sugarcane or sorghum stalks are
crushed to extract their juice. It is used as a biofuel for the production of heat, energy, and
electricity, and in the manufacture of pulp and building materials.
Agave bagasse is a similar material that consists of the tissue of the blue agave after extraction
of the sap.
For every 10 tonnes of sugarcane crushed, a sugar factory produces nearly three tonnes of wet
bagasse. Since bagasse is a by-product of the cane sugar industry, the quantity of production in
each country is in line with the quantity of sugarcane produced.
The high moisture content of bagasse, typically 40–50 percent, is detrimental to its use as a
fuel. In general, bagasse is stored prior to further processing. For electricity production, it is
stored under moist conditions, and the mild exothermic process that results from the
degradation of residual sugars dries the bagasse pile slightly. For paper and pulp production, it
is normally stored wet in order to assist in removal of the short pith fibres, which impede the
paper making process, as well as to remove any remaining sugar.
A typical chemical analysis of washed and dried bagasse might show.

 Cellulose 45–55 percent

 Hemicellulose 20–25 percent
 Lignin 18–24 percent
 Ash 1–4 percent
 Waxes <1 percent
Bagasse is a heterogeneous material containing around 30-40 percent of "pith" fibre, which is
derived from the core of the plant and is mainly parenchyma material, and "bast", "rind", or
"stem" fibre, which makes up the balance and is largely derived from sclerenchyma material.
These properties make bagasse particularly problematic for paper manufacture and have been
the subject of a large body of literature.
FIG2.1: SUGARCANE WASTE OR BAGASSE
 2.2:POTATO PEEL WASTE
Potatoes are one of the most important agricultural crops for human consumption after wheat
rice and maize with 376 million tons produced in 2013. In developed countries up to 69.5% (in
2012) of total produced potatoes are processed. Potatoes are usually peeled during processing
and production losses in a form of potato peel waste (PPW) can vary from 15 to 40%,
depending on the peeling method Each year huge quantities of PPW as a by-product remain
after industrial potato processing. Abrasion peeling is typical for chips production, whereas
steam peeling is used for dehydrated and frozen potato products . Steam peelers are compact
and generate less product losses, but require high investment and operation costs. Because of
that, steam peeling is reasonable when high quantities of product (from 8 to 20 t h-1) have to be
peeled in limited space and appearance of brown ring (also known as heat ring, or cooking ring)
does not cause problems for final product . Brown ring occurs due to tissue damage and
enzyme-catalyzed phenolic oxidation reaction . It is reported, that chemical peeling using NaOH
could replace steam peeling to avoid heat-ring . Otherwise abrasion peeling is to be used. PPW
is not suitable for non-ruminants without further treatment because it is too fibrous to be
digested (Birch et al., 1981), but as an inexpensive by-product it contains a large quantity of
starch, nonstarch polysaccharides, lignin, polyphenols, protein and small amount of lipids. This
makes it a cheap and valuable base material for extraction of valuable products (such as natural
antioxidants, dietary fibre, biopolymers, etc.) and fermentation processes . Many studies were
made and articles were written on PPW application possibilities in order to minimize industrial
waste amount and find suitable application for PPW as a by-product. The aim of the present
study is to summarize the available literature on possible industrial PPW utilization methods and
highlight its application possibilities in food production.

TABLE 2.1:CHEMICAL COMPOSITION OF RAW POTATO PEEL, g 100 g-1

Compound Minimum and maximum Average content Source

values

water 83.3-85.1 84.2 Potatoes, raw, skin,

s.a.Arapoglou et al., 2009

Protein 1.2-2.3 1.8 Potatoes, raw, skin,

s.a.Arapoglou et al., 2009

Total lipid 0.1-0.4 0.3 Potatoes, raw, skin,

s.a.Arapoglou et al., 2009

Total 8.7-12.4 10.6 Potatoes, raw, skin,

carbohydrate s.a.Arapoglou et al., 2009

starch 7.8 Arapoglou et al., 2009

Total dietary fibre 2.5 Potatoes, raw, skin, s.a.

2.3: MANGO LEAF WASTE

Mango is processed to a maximum extent, thereby producing high quality of solid and liquid
wastes. Solid wastes, stones, stalks, trimmings and fibrous materials are obtained during the
preparation of raw material. This contributes about 40 to 50% of total fruit waste out of which, 5
to 10% is pulp waste and 15 to 20% is kernel (Anonymous, 2004; Madhukara et al., 1993; Maini
et al., 2000; Pandey et al., 2000). Liquid waste is the waste material that comes out of a factory
after washing of fruits, packaging, blanching, cooling and plant and machinery clean up and so
on. Utilization of this mango waste is both a necessity and a challenge. If a factory is processing
five tons of Totapuri mangoes per hour, about six tons of peel would be available as waste per
day of 8 h work. Approximately, 0.4 to 0.6 million tons of mango peel is generated annually in
India (Anonymous, 2004). This waste is either used as cattle feed or dumped in open areas,
where it adds to environmental pollution. The use of mango peel as a source of pectin and fibre
production also has been reported (Pandia et al., 2004). Grohmann et al. (1994; 1995; 1996;
1998) previously reported ethanol production from orange peel. Ethanol production from banana
(Manikandan et al., 2008) and pineapple peels (Ban-koffi and Han, 1990) were also
investigated. Mango peel is difficult to decompose, as it takes a very long time, because of its
complex composition. Suitability of mango peel for biogas production was investigated by
Madhukara et al. (1993). However, ethanol fermentation from fruit and vegetable wastes like
mango peel appears to give better returns. The presence of high amount of reducing sugars in
dried and fresh mango peel prompted us to make an attempt to utilize it as a raw material for
ethanol production and development of cheap medium. As far as we know, this is the first report
of its kind on ethanol production from mango peel.

TABLE 2.2: COMPOSITION OF FRESH AND DRIED MANGO PEEL

Content Fresh Mango peel Dried Mango peel

Moisture 4.3 ± 0.5 10 ± 1.2

Total solids 25.6 ± 4.6 70.5 ± 2.7

Reducing sugar 7.0 ± 1.8 30 ± 2.5

Non-reducing sugar 5.9 ± 0.4 4.3 ± 0.5

Protein 3.5 ± 0.5 4.0 ± 0.8

Cellulose and Lignin 25.2 ± 2.0 23 ± 1.2

2.4:BAKERS YEAST(Saccharomyces cerevisiae)
Baker's yeast is the common name for the strains of yeast commonly used in baking bread and
bakery products, serving as a leavening agent which causes the bread to rise (expand and
become lighter and softer) by converting the fermentable sugarspresent in
the dough into carbon dioxide and ethanol. Baker's yeast is of the species Saccharomyces
cerevisiae, and is the same species (but a different strain) as the kind commonly used in
alcoholic fermentation, which is called brewer's yeast. Baker's yeast is also a single-cell
microorganism found on and around the human body.
The use of steamed or boiled potatoes, water from potato boiling, or sugar in a bread dough
provides food for the growth of yeasts; however, too much sugar will dehydrate them. Yeast
growth is inhibited by both salt and sugar, but more so by salt than sugar. Some sources
say fats, such as butter and eggs, slow down yeast growth others say the effect of fat on dough
remains unclear, presenting evidence that small amounts of fat are beneficial for baked bread
volume.
Saccharomyces Exiguus (also known as S. minor) is a wild yeast found on plants, fruits, and
grains that is occasionally used for baking; however, in general, it is not used in a pure form but
comes from being propagated in a sourdough starter.
Baker's yeast is available in a number of different forms, the main differences being the
moisture contents. Though each version has certain advantages over the others, the choice of
which form to use is largely a question of the requirements of the recipe at hand and the training
of the cook preparing it. Dry yeast forms are good choices for longer-term storage, often lasting
more than a year at room temperatures without significant loss of viability. [15] In general, with
occasional allowances for liquid content and temperature, the different forms of commercial
yeast are considered interchangeable.
2.4.1:TYPES OF YEAST

 Cream yeast is the closest form to the yeast slurries of the 19th century, in essence being a
suspension of yeast cells in liquid, siphoned off from the growth medium. Its primary use is
in industrial bakeries with special high-volume dispensing and mixing equipment, and it is
not readily available to small bakeries or home cooks.
 Compressed yeast is, in essence, cream yeast with most of the liquid removed. It is a soft
solid, beige in color, and best known in the consumer form as small, foil-wrapped cubes
of cake yeast. It is also available in a larger-block form for bulk usage. It is highly
perishable; though formerly widely available for the consumer market, it has become less
common in supermarkets in some countries due to its poor keeping properties, having been
superseded in some such markets by active dry and instant yeast. It is still widely available
for commercial use, and is somewhat more tolerant of low temperatures than other forms of
commercial yeast; however, even there, instant yeast has made significant market inroads.
 Active dry yeast is the form of yeast most commonly available to non-commercial bakers in
the United States. It consists of coarse oblong granules of yeast, with live yeast cells
encapsulated in a thick jacket of dry, dead cells with some growth medium. Under most
conditions, active dry yeast must first be proofed or rehydrated. It can be stored at room
temperature for a year, or frozen for more than a decade, which means that it has better
keeping qualities than other forms, but it is generally considered more sensitive than other
forms to thermal shock when actually used in recipes.

A single grain of active dry yeast. The numbered ticks on the scale are 230 µm apart

 Instant yeast appears similar to active dry yeast, but has smaller granules with substantially
higher percentages of live cells per comparable unit volumes. It is more perishable than
active dry yeast but also does not require rehydration, and can usually be added directly to
all but the driest doughs. In general, instant yeast has a small amount of ascorbic
acid added as a preservative. Some producers provide specific variants for doughs with
high sugar contents, and such yeasts are more generally known as osmotolerant yeasts.
 Rapid-rise yeast is a variety of dried yeast (usually a form of instant yeast) that is of a
smaller granular size, thus it dissolves faster in dough, and it provides greater carbon
dioxide output to allow faster rising. There is considerable debate as to the value of such a
product; while most baking experts believe it reduces the flavor potential of the finished
product, Cook's Illustrated magazine, among others, feels that, at least for direct-rise
recipes, it makes little difference. Rapid-rise yeast is often marketed specifically for use
in bread machines.
 Deactivated yeast is dead yeast which has no leavening value and is not interchangeable
with other yeast types. Typically used for pizza and pan bread doughs, it is used at a rate of
0.1% of the flour weight, though manufacturer specifications may vary. It is a powerful
reducing agent used to increase the extensibility of a dough.
For most commercial uses, yeast of any form is packaged in bulk (blocks or freezer bags for
fresh yeast; vacuum-packed brick bags for dry or instant); however, yeast for home use is often
packaged in pre-measured doses, either small squares for compressed yeast or sealed packets
for dry or instant. For active dry and instant yeast, in general a single dose (reckoned for the
average bread recipe of between 500 g and 1000 g of dough) is about 2.5 tsp (~12 mL) or about
7 g (1⁄4 oz), though comparatively lesser amounts are used when the yeast is used in a pre-
ferment. In general, a yeast flavor in the baked bread is not noticeable when the bakers'
percent of added yeast is less than 2.5%.
 3.PRETREATMENT TECHNOLOGIES
The increasing problem of the CO2 emissions besides some energy security concerns has
strengthened the interest in alternative,nonpetroleum-based sources of energy. Biomass is the
only suitable and renewable primary energy resource than can provide alternative transportation
fuels such as bioethanol or biodiesel. Current production of bioethanol relies on ethanol from
starch
and sugars but there has been considerable debate about its sustainability. Bioethanol
produced from lignocellulosic biomass is an interesting alternative since lignocellulosic raw
materials do not compete with food crops and they are also less expensive than conventional
agricultural feedstocks.The biological conversion of different lignocellulosicfeedstocks such as
forest and agricultural residues, or lignocellulosic crops dedicated to ethanol offers numerous
benefits but its development is still hampered by economic and technical obstacles.Some of the
most important factors to reduce ethanol production cost are: an efficient utilization of the raw
material to obtain high ethanol yields, high productivity, high ethanol concentration in the
distillation feed, and also process integration in order to reduce the energy demand.

3.1:FACTORS LIMITING ENZYMATIC HYDROLYSIS

The pretreatment is a necessary step to alter some structural characteristics of lignocellulose,
increasing glucan and xylan accessibility to the enzymatic attack. As it has been mentioned,
these
structural modifications of the lignocellulose are highly dependent on the type of pretreatment
employed and have a great effect on the enzymatic hydrolysis.

Main factors that influence the enzymatic hydrolysis of cellulose in lignocellulosicfeedstocks can
be divided in two groups: enzyme-related and substrate-related factors, though many of them
are interrelated during the hydrolysis process. Composition of the liquid fraction and solid
process streams resulting from different pretreatment approaches can be widely different.
These differences will have a great influence on the requirements for effective enzymatic
saccharification in subsequent processing step.The reduction of pretreatment severity is
sometimes required
to reduce economic cost. Low severity factor results in less sugar-release and consequently
higher amount and different types of enzymes will be required to achieve high sugar yields from
both
cellulose and hemicellulose fraction. In this context, development of hemicellulases and other
accessory enzymes needed for complete degradation of lignocellulose components has
become an
important issue.

1.CELLULOSE CYRSTALLINITY (CRYSTALLINITY INDEX ,Crl)

Degree of polymerization and cellulose crystallinity have been considered as important factors
in determining the hydrolysis rates of relatively refined cellulosic substrate.In some studies
wherein crystallinity was suggested to be important, the lignocellulosic materials were
mechanically pretreated, therefore any decrease in crystallinity was accompanied by an
alteration of other substrate characteristics such as particle size reduction or increase in
available surface area.
It has been observed that pretreatment of lignocellulosics improves its hydrolysability but in
some cases increases the CrI of the cellulose fraction. This fact has been suggested to be due
to
the removal or reduction of more easily available amorphous cellulose after pretreatments such
steam explosion. In contrast, high pH pretreatments have been shown to have less effect and
even reduced biomass cristallinity in some instances.

2.CELLULOSE DEGREE OF POLYMERISATION (DP, NUMBER OF GYLCOSYL RESIDUES

PER CELLUOLSE CHAIN)

Degree of polymerization is essentially related to other substrate characteristics, such as

cristallinity. Although the role of glucan chain length is not definitively known, it is believed to
affect
cellulose hydrolysis. Depolymerization depends on the nature of cellulosic substrate. In the
enzymatic hydrolysis, endoglucanases cut at internal sites of the cellulose chains, preferentially
less ordered, being primarily responsible for decreasing degree of polymerization of cellulosic
substrates. However, regardless the substrate being attacked there seems to be a „„leveling off”
of the cellulose DP, correlated with the increased recalcitrance of the residual crystalline
cellulose.The effect of different pretreatments on cellulose chain length has been studied
showing reduced degree of polymerization in solids prepared by different pretreatments
suggesting that xylan removal had a more severe impact on cellulose chain length than lignin
removal.

3.SUBSTRATES AVAILABLE SURFACE AREA (PORE VOLUME)

Accessibility of the substrate to the cellulolytic enzymes is one of the major factors influencing
hydrolysis process. Thus, one of the main objectives of the pretreatment is to increase the
available
surface area for the enzymatic attack.
4.LIGNIN BARRIER (CONTENT AND DISTRIBUTION)

The presence of lignin and hemicellulose difficults the access of cellulase enzymes to cellulose
difficult, thus reducing the efficiency of the hydrolysis. Lignin limits the rate of enzymatic
hydrolysis by acting as a physical barrier, preventing the digestible parts of the substrate to be
hydrolyzed . Besides, lignin appears to reduce cellulose hydrolysis by non-productively binding
cellulolytic enzymes. Different strategies have been studied recently to overcome the non-
productive adsorption of cellulase to lignin such as alkali extraction and addition of protein (e.g.
BSA) or other additives (e.g. poly ethylene glycol). Although the use of additives introduces an
additional cost to the ethanol production process, significant benefits could be achieved by
improving the enzymatic hydrolysis step.Some pretreatments have been reported to produce
different effects such as melting and lignin relocation (steam explosion) or disruption of lignin–
carbohydrates linkages (AFEX).

5.HEMICELLULOSE CONTENT

Removal of hemicellulose increases the mean pore size of the substrate and therefore
increases the accessibility and the probability of the cellulose to become hydrolysed. On the
other hand, hemicellulosic sugars recovery in the pretreated solids would be interesting to
obtain higher total fermentable sugar production. In this case enzymatic requirements for
hemicellulosic modification must be taken into account . Degree of acetylation in the
hemicellulose is another important factor because lignin and acetyl groups are attached to the
hemicellulose matrix and may hinder polysaccharide breakdown.

6.FEED STOCK PARTICLE SIZE

There is some evidence to support that reduction of particle size increases specific surface area
and subsequently the accessibility of cellulose to the enzymes.

7.POROSITY

The pore size of the substrate in relation to the size of the enzymes is the main limiting factor in
the enzymatic hydrolysis of lignocellulosic biomass. Cellulases can get trapped in the pores if
the internal area is much larger than the external area which is the case for many
lignocellulosicmaterials . An increase of porosity in pretreatment processes can significantly
improve the hydrolysis.

8.CELL WALL THICKNESS

The waxy barrier comprising grass cuticle and tree bark impedes penetration of enzymes; even
milled, plant stems and woody tissues limit liquid penetration by their nature.

9.CHANGE IN ACCESSIBILITY IN CONVERSION

The role of glucan accessibility and its change with conversion has been debatable, with a few
studies showing that glucan accessibility becomes limiting with conversion and a few others
showing not significant decrease of accessibility with conversion or even no change at all.
3.2: PRETREATMENT TECHNOLOGIES FOR LIGNOCELLULOSIC
BIOMASS
A multitude of different pretreatment technologies have been suggested during the last
decades. They can be classified into biological, physical, chemical and physico-chemical
pretreatments.

1.BIOLOGICAL PRETREATMENTS

Fungal pretreatment has been previously explored, this environmentally friendly approach has
received renewed attention as a pretreatment method for enhancing enzymatic saccharification
of lignocellulosic biomass in ethanol production processes. Biological pretreatments employ
microorganisms mainly brown, white and soft-rot fungi which degrade lignin and hemicellulose
and very little of cellulose, more resistant than the other components.Lignin degradation by
white-rot fungi, the most effective for biological pretreatment of lignocellulosic materials, occurs
through the action of lignin-degrading enzymes such as peroxidases and laccases.Biological
pretreatment by white-rot fungi has been combined with organosolv pretreatment in an ethanol
production process by simultaneous saccharification and fermentation (SSF) from beech wood
chips.Results from other recent studies have shown that fungal pretreatment of wheat straw for
10 days with a high lignin-degrading and low cellulose-degrading fungus (fungal isolate RCK-1)
resulted in a reduction in acid loading for hydrolysis, an increase in the release of fermentable
sugars. a
reduction in the concentration of fermentation inhibitors. Ethanolyield and volumetric
productivity with Pichiastipitis were 0.48 g/g and 0.54 g/L h, respectively.An evaluation of
biological pretreatment of sugarcane trash using eight different bacteria and fungi was
performed on the basis of quantitative changes in the components of the sugarcane trash, the
production of the cellulase enzyme complex, total protein and the release of reducing sugars by
different bioagents as well as

the interaction among different chemical parameters affecting the pretreatment .In this case, the
microbial pretreatment of trash increased accessibility of sugars for enzymatic hydrolysis.such
processes offer advantages such as low-capital cost, low energy, no chemicals requirement,
and mild environmental conditions. However, the main drawback to develop biological methods
is the low hydrolysis rate obtained in most biological materials compared to other technologies.

2.PHYSICAL PRETREATMENTS

2.1.MECHANICAL COMMINUTION

The objective of the mechanical pretreatment is a reduction of particle size and cristallinity of
lignocellulosic in order to increase the specific surface and reduce the degree of polymerization.
This can be produced by a combination of chipping, grinding or milling
depending on the final particle size of the material (10–30 mm after chipping and 0.2–2 mm
after milling or grinding)Different milling processes (ball milling, two-roll milling, hammer milling,
colloid milling and vibro energy milling) can be used to improve the enzymatic hydrolysis of
lignocelullosic
materials (Taherzadeh and Karimi, 2008). The power requirement of this pretreatment is
relatively high depending on the final particle size and the biomass characteristics. Taking into
account the
high energy requirements of milling and the continuous rise of energy prices, it is likely that this
process is not economically feasible.

2.2.EXTRUSION

In extrusion, the materials are subjected to heating, mixing and shearing, resulting in physical
and chemical modifications during the passage through the extruder. Screw speed and barrel
temperature are believed to disrupt the lignocellulose structure causing defibrillation, fibrillation
and shortening of the fibers, and, in the end, increasing accessibility of carbohydrates to
enzymatic attack The different bioreactor parameters must be taken into account to achieve the
highest efficiency in the process. In recent studies application of enzymes during extrusion
process is being considered as a promising technology for ethanol production.

3. CHEMICAL PRETREATMENTS

3.1 ALKALI PRETREATMENTS

Alkali pretreatments increase cellulose digestibility and they are more effective for lignin
solubilization, exhibiting minor cellulose and hemicellulose solubilization than acid or
hydrothermal processes. It cause less sugar degradation than acid pretreatment and it was
shown to be more effective on agricultural residues than on wood materials.Sodium, potassium,
calcium and ammonium hydroxides are suitable alkaline pretreatments. NaOH causes swelling,
increasing the internal surface of cellulose and decreasing the degree of polymerization and
cristallinity, which provokes lignin structure disruption .NaOH has been reported to increase
hardwood digestibility from 14% to 55% by reducing lignin content from 24–55% to 20%.Lime
pretreatment removes amorphous substances such as lignin, which increases the crystallinity
index. Lignin removal increases enzyme effectiveness by reducing non-productive adsorption
sites for enzymes and by increasing cellulose accessibility.Lime also removes acetyl groups
from hemicellulose reducing steric hindrance of enzymes and enhancing cellulose
digestibility.Lime has been proven successfully at temperatures from 85–150 C and for 3–13 h
with corn stover or poplar wood. Pretreatment with lime has lower cost and less safety
requirements compared to NaOH or KOH pretreatments and can be easily recovered from
hydrolysate by reaction with CO2.Addition of an oxidant agent (oxygen/H2O2) to alkaline
pretreatment (NaOH/Ca(OH)2) can improve the performance by favoring lignin removal.Ethanol
yields of 0.33 g/g have been obtained in simultaneous saccharification and cofermentation
(SSCF) processes with Escherichia coli FBR5 from wheat straw pretreated with alkali peroxide.

3.2 ACID PRETREATMENTS

The main objective of the acid pretreatments is to solubilize the hemicellulosic fraction of the
biomass and to make the cellulose more accessible to enzymes.Equipment corrosion problems
and acid recovery are important drawbacks when using concentrated acid pretreatments. The
high operational and maintenance costs reduce the interest of applying the concentrated acid
pretreatment at commercial scale.Diluted acid pretreatment appears as more favourable
method for industrial applications.Different types of reactors such as percolation, plug flow,
shrinking-bed, batch and countercurrent reactors have been applied for pretreatment of
lignocellulosicmaterials . It can be performed at high temperature (e.g. 180 C) during a short
period of time; or at lower temperature (e.g. 120 C) for longer retention time (30–90 min). It
presents the advantage of solubilizing hemicellulose, mainly xylan, but also converting
solubilized hemicellulose to fermentable sugars. Nevertheless, depending on the process
temperature, some sugar degradation compounds such as furfural and HMF and aromatic lignin
degradation compounds are detected, and affect the microorganism metabolism in the
fermentation step.Organic acids such as fumaric or maleic acids are appearing as alternatives
to enhance cellulose hydrolysis for ethanol production. In this context, both acids were
compared with sulfuric acid in terms of hydrolysis yields from wheat straw and formation of
sugar degradation compounds during pretreatment. Results showed that organic acids can
pretreat wheat straw with high efficiency although fumaric acid was less effective than maleic
acid. Furthermore, less amount of furfural was formed in the maleic and fumaric acid
pretreatments than with sulfuric acid..

4. PHYSICO CHEMICAL PRETREATMENT

4.1 Steam explosion or autohydrolysis

This method of pretreatment without the use of any catalyst is promising

and the biomass fractionates to yield levulinic acid, xylitol and alcohols In this method the
biomass is heated using highpressure steam (20-50 bar), 160-290 degree celcius) for a few
minutes; the reaction is then stopped by sudden decompression to atmospheric pressure.
When steam is allowed to expand within the lignocellulosic matrix it separates the individual
fibers. The high recovery of xylose (45-65%) makes steam-explosion pretreatment economically
attractive
4. 2 Liquid hot water method

The liquid hot water method uses compressed hot liquid water (at pressure above saturation
point) to hydrolyze the hemicelluloses . It is a hydrothermal pretreatment method which releases
high fraction of hemicellulosic sugars in the form of oligomers. The treatment generally occurs
at temperatures of 170-230 degree celcius and pressures above MPa for 20 min. it produce
small amounts of undesired degrading compounds such as furfural, carboxylic acid, that are very
toxic to ethanol fermentation as they inhibit microbial growth. As xylose recovery is relatively
high (88-98%), and no acid or chemical is required, it is an environmentally attractive and
economically interesting method.

4.3ULTRASOUND PRETREATMENT

The effect of ultrasound on lignocellulosic biomass have been employed for extracting
hemicelluloses, cellulose and lignin but less research has been addressed to study the
susceptibility of lignocellulosic materials to hydrolysis.Higher enzymatic hydrolysis yields after
ultrasound pretreatment could be explained because cavitation effects caused by
introduction of ultrasound field into the enzyme processing solution greatly enhance the
transport of enzyme macromolecules toward the substrate surface. Furthermore, mechanical
impacts, produced by the collapse of cavitation bubbles, provide an important benefit of opening
up the surface of solid substrates to the action of enzymes, in addition, the maximum effects of
cavitation occur at 50 C, which is the optimum temperature for many enzymes.

4. ENZYMATIC HYDROLYSIS
Enzymatic hydrolysis of cellulose is carried out by cellulose enzymes ,which are highly
specific.

There are three major steps involved in the hydrolysis in process .

4.1 Endoglu-canases which attacks region of low crystallinity in the cellulose fiber,creating free
chain ends.

4.2 Exoglucanases or cellobiohydrolase Which degrades the molecule further by removing

cellobiose units from free chains .

4.2 BETA gluocose which hydrolyzes cellobiose to produce glucose in addition to three major
groups of cellulase enzymes ,there are also a number of ancillary enzymes that attack
hemicellulose ,such as glucuronidase, acetylesterasr, xylanase, beta xylosidase
,galactomannase and glucomannanaseduring hydrolysis cellulose is degraded by the
thecelluases to reducing sugars ,which can be fermented by yeast or bacteria to ethanol .
4.2 utility cost of enzymatic hydrolysis is low compared to acid or alkaline hydrolysis because
ezyme hydrolysis is usually conducted at mild conditions (PH 4.8 and temperature 45-50 c) and
doesn‟t have corrosion problem.

4.3 Both bacteria and fungi can produce cellulase s for hydrolysis of
lignocellulosicmaterials,these microorganism can be anaerobic/aerobic ,mesophilic /themophilic
.

4.4 one type of bacteria known as filamentous fungus fusariumoxysporumposses ability to

produce ethanol by simultaneous saccharification and fermentation of cellulose.

4.5 factors that affect the enzymatic hydrolysis of cellulose include substrate ,cellulase activity
and reaction conditions.

4.6 substrate concentration is one of the main factors that affect the yield and initial rate of
enzymatic hydrolysis of celluolose.at low substrate ,an increase of substrate concentration
normally results in an incease of the yield and reaction rate of hydrolysis.

4.7 high subsrate concentration can cause substrate inhibition which subsequently lowers the
rate of hydrolysis and extent depends upon the ratio of the total substrate to total enzyme .

4.8 incerasing the dosage of cellulase in the process can inhance the yield and rate of
hydrolysis.

4.9 cellulose activity decrease during the hydrolysis.

4.10 irreversible adsorption of cellulase on cellulose is partially responsible for this activation.

4.11 in the process reducing sugars produced inn cellulose or sacchrification is simultaneously
fermented to ethanol.
 5.Fermentation
The saccharified biomass is used for fermentation by several microorganisms. But the industrial
utilization of lignocelluloses for bioethanol production is hindered by the lack of ideal
microorganisms which can efficiently ferment both pentose and hexose sugars . For a
commercially viable ethanol production method,an ideal microorganism should have broad
substrate utilization,high ethanol yield and productivity, should have the ability to withstand high
concentrations of ethanol and high temperature, should be tolerant to inhibitors present in
hydrolysate and have cellulolytic activity. Genetically modified or engineered microorganisms
are thus used to achieve complete utilization of the sugars in the hydrolysate and better
production benefits. The processes usually employed in the fermentation of
lignocellulosichydrolysate are simultaneous saccharification and fermentation (SSF) and
separate hydrolysis and fermentation (SHF).Conventionally or traditionally the SHF process has
been employed but SSF is superior for ethanol production since it can improve ethanol yields by
removing end product inhibition and eliminate the need for separate reactors. It is also cost
effective but difference in optimum temperature conditions of enzyme for hydrolysis and
fermentation poses some limitations. The higher ethanol yield coefficient from SSF would be
partially due to more conversion of xylose to xylitol under the SSF conditions.ssf is better
alternative to SHF as per studied done. Some native or wild type microorganismused in the
fermentation are S. cerevisiae, ,Zymomonasmobilis, known yeast and bacteria employed in
ethanol production from hexoses are S. cerevisiae and Z. mobilis respectively . But S.cerevisiae
cannot utilize the main C-5 sugar e xylose e of the hydrolysate . Native organisms such as
Pichia and Candida species can be used in place of S. cerevisiae and they can utilize xylose but
their ethanol production rate is at least fivefold lower than that observed with S.
cerevisiaediffrent microorganisms have shown different yields of ethanol depending on their
monomer utilization .Genetic engineering has been employed to develop the various aspects of
fermentation from higher yield to better and wide substrate utilization to increased recovery
rates. Fermentation is series of chemicai reaction that convert sugar to ethanol.the fermentation
reaction is caused by(eg; saccharomyces cerevisiae) or bacteria which feed on the
sugar.ethanol and carbon dioxide are produce as sugar is consumed.

Straw eco-industerial multiple production pattern with biofuel for leading product taking low
pressure steam explosion and solid-state fermentation as core technologies.
6. DISTILLATION PROCESS

By using simple distillation method the fermentation product from glucose fermentation and
pentose fermentation is called ethanol broth . in this step ethanol is separated from the other
component in the broth. A final dehydration step removes any remaining water from the ethanol.
By this way we obtained ethanol.

8.PICTURE FROM OUR EXPERIMENT

ETHANOL OBTAINED AFTER DISTILLATION OF SUGARCANE BAGASSES OR ETHANOL
SAMPLE OBTAINED

SAMPLE OBTAINED AFTER FERMENTATION

9. BIOETHANOL PROPERTIES

Bioethanol , raw sugar beet, corn , wheat and other woody plants such as sugar,starch oe
cellulose to gasoline and concise obtained by fermentation of agricultural products is an
alternative fuel that is blended in specific proportions. Bioethanol is a clear,colourless liquid with
a characteristics odour. It is a high octane fuel (113) is the boiling point of 78.5 degree celcius,
the freezing point of -114.1 degree celcius. Bioethanol 20 degree celcius , density 0.789gm/ml.
Internal combustion engines used in blending amount of 10% without the need for any
modification.

9.1 BIOETHANOL IS BLENDED WITH GASOLINE

 Bioethanol used in order to increase the octane no in the fuel benzene, methyl tertary
butyl ether(MTBE) as friendly alternative to carcinogenic substances
 Bioethanol fuel according to the blending ratio improves engine performance by
providibg an octane no increase of 2-3 % points
 Bioethanol prevents freezing, the engine keeps the cooler and cleaner nozzle.

10.USES OF BIOETHANOL

10.1.TRANSPORTATION SECTOR
  Mixed with gasoline
 Diesel as an additive in motors
 Recent technological vehicles(hybrid ,fuel cell)
 In agriculture

11.APPLICATIONS OF BIOETHANOL

11. 1 HOME APPLICATIONS

 In oven
 Lightening
 In heating and cooling devices
 Storage of food (cooling)

11.2 CHEMICAL SECTOR

 In ethylene production
 In hydrogen production
 In glycol ethers
 Ethyl acrylate
 Acetic acid
 Ethyl acetate
 Acetaldehyde
 Ethyl ether

12. BIOETHANOL WITH ENVIRONMENTAL BENEFITS

 Bioethanol fuel to increase the oxygen level in the easiest way.

 To increase the oxygen level of the fuel , providing a more efficient fuel combustion
reduces harmful gases in exhaust.
 When used with the aim to increase the octane bioethanol fuel, benzene, methyl tertary
butyl ether(MTBE) as friendly alternative carcinogenics substances.
 Bioethanol reduces exhaust emissions.
 Bioethanol blends, which leads to a reduction of the ozone layer allows substantially
decrease in hydrocarbon emissions.
 High level bioethanol mixtures provides upto 20% reduction in nitrogen oxide emissions.

13. ADVANTAGES

 Exhaust gases of ethanol are much cleaner, it burns more cleanly(more complete
combustion).
 The use of ethanol blended fuels such as E85 (85%ethanol and 15% gasoline) can
reduce the net emission of green house gases by as much as 37.1%,which is a
significant amount.
 Posistive energy balance.
  It is a carbon natural that is the carbon dioxide released in the bioethanol production
process is the same amount as the one crops previously absorbed during
photosynthesis.
 You can use any plants for production of bioethanol, it only has to contain sugar and
starch.
 The best choice is the sugarcane, but you can use potatoes ,wheat etc.
 Ethanol is considered a renewable energy resource because it is primarily a result of
conversion of the sun‟s energy into usable energy creation of ethanol starts with
photosynthesis, which causes feed stocks, such as sugarcane, to grow. These particular
feedstocks are processed into ethanol.
 Its reduces green house gases.
 It reduces the amount of high octane additives.
 The fuel spills are more easily biodegraded or diluted to non toxic concentrations.

14.DISADVANTAGES

 Carbon emissions during production of bioethanol huge amount of carbondioxide is

released which makes its ecological effectiveness close to zero.
 Pure ethanol is also difficult to vapourise which can make starting a car in cold weather
difficult. And that is why most fuels retain atleats a snall amount of petrol sucha as E85
cars with 85%ethanol and 15% gasoline.
 It is quite expensive to setup a biofuel laboratory.
 It leads to biodiversity due to the fact that a large amount of arable lands is required to
grow these crops and could see some natural habitats destroyed including reinforests.
 Bioethanol production demands strong technical knowledge for effective production and
also to avoid excess emissions.