Renewable and Sustainable Energy Reviews 71 (2017) 475–501

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Renewable and Sustainable Energy Reviews

journal homepage: www.elsevier.com/locate/rser

Bioethanol production from renewable sources: Current perspectives and MARK

technological progress
⁎
H. Zabeda,b, , J.N. Sahuc,d,e,⁎⁎, A. Suelya, A.N. Boycea, G. Faruqf
a
Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia
b
Department of Microbiology, Primeasia University, 12, Kemal Ataturk Avenue Banani, Dhaka 1213, Bangladesh
c
Department for Management of Science and Technology Development, Ton Duc Thang University, Ho Chi Minh City, Vietnam
d
Faculty of Applied Sciences, Ton Duc Thang University, Ho Chi Minh City, Vietnam
e
Petroleum and Chemical Engineering Program Area, Faculty of Engineering, Universiti Teknologi Brunei, Tungku Gadong, P.O. Box 2909, Brunei
Darussalam
f
Bangladesh Agricultural Research Institute, Joydebpur, Gazipur 1701 Bangladesh

A R T I C L E I N F O A BS T RAC T

Keywords: Bioethanol is an attractive biofuel having potential for energy security and environmental safety over fossil fuels.
First generation bioethanol To date, numerous biomass resources have been investigated for bioethanol production, which can broadly be
Second generation bioethanol classified into sugars, starch and lignocellulosic biomass. However, conversion of biomass into ethanol varies
Lignocellulosic biomass considerably depending on the nature of feedstock, primarily due to the variation in biochemical composition,
Zymomonas mobilis
and so, only a few feedstocks have been exploited commercially. In recent years, the conversion process of
Saccharomyces cerevisiae
Starchy crop
biomass has been improved significantly, even though most of these achievements are yet to be implemented in
commercial facility. All the major steps in a typical conversion process, particularly fermentation of sugars that
is the common step for all biomass, are greatly influenced by microorganisms. A traditional yeast,
Saccharomyces cerevisiae, and a bacterial species, Zymomonas mobilis, are widely used in the ethanol
fermentation technology. Many factors affect ethanol production process, and the final yield is directly
associated with the optimum conditions of these attributes. This review paper presents an overview on the first
and second generation bioethanol production with a particular attention to the potential of various biomass
sources, technological approaches, role of microorganisms and factors affecting ethanol production process.

1. Introduction
Energy security and environmental safety are two major issues in
the current world that have boosted the demand for an alternative and
eco-friendly energy source. It has been anticipated that fossil fuels
reserve will be exhausted by next 40–50 years due to rapid increase in
the consumption rate of these non-renewable fuels [1]. More impor-
tantly, the burning of fossil fuels contributes to the emissions of
greenhouse gases as well as global warming that causes climate change,
rise in sea level, loss of biodiversity and urban pollution [2–4].
Bioethanol is one of the most promising alternatives to fossil fuels,
which can be produced from various renewable sources rich in
carbohydrates. Many countries, such as USA, Brazil, China, Canada
and several EU member states have already proclaimed commitments
to bioethanol programs as attempts to reduce the dependence on fossil
fuels, where the former two countries have shown the largest commit-
ments thus far. In response to the current demand, global bioethanol
production has been increased over time as shown in Fig. 1a. United
states produces the highest amount of ethanol, which has been
estimated to be more than half of the total global ethanol produced
in 2015 [5]. Total ethanol production in the USA has increased
dramatically from 175 million gallons in 1980 to 14810 million gallons
in 2015 (Fig. 1b) [5].
Ethanol (C2H5OH) has been earmarked as a promising energy
source over gasoline (C7H17) due to having several advantageous
properties. Even though one liter of ethanol affords 66% of the energy
provided by the same amount of gasoline, the former has a higher
octane number (106–110) than the latter (91–96), which enhances the
performance of gasoline when blended with ethanol [6]. The higher
octane level of ethanol also allows it to be burnt at a higher compres-
sion ratio with shorter burning time, resulting in a lower engine knock.
In addition, ethanol has a higher evaporation enthalpy (1177 kJ/kg at

⁎
Corresponding author at: Department of Microbiology, Primeasia University, 12, Kemal Ataturk Avenue Banani, Dhaka 1213, Bangladesh.
⁎⁎
Corresponding author at: Department for Management of Science and Technology Development, Ton Duc Thang University, Ho Chi Minh City, Vietnam.
E-mail addresses: zabedctgbd@yahoo.com (H. Zabed), jay_sahu@yahoo.co.in, jayanarayansahu@tdt.edu.vn (J.N. Sahu).

http://dx.doi.org/10.1016/j.rser.2016.12.076
Received 20 October 2015; Received in revised form 5 December 2016; Accepted 12 December 2016
Available online 05 January 2017
1364-0321/ © 2016 Elsevier Ltd. All rights reserved.
H. Zabed et al.
Fig. 1. (a) Global bioethanol production by country or region during last 10 years; (b)
Ethanol production by the USA during 1980–2015 (Data were collected from [5]).
60 °C) than gasoline (348 kJ/kg at 60 °C) and a higher laminar flame
speed (around 33 and 39 cm/s at 100 kPa and 325 K for gasoline and
ethanol, respectively) [7–10]. The higher heat of vaporization of
ethanol (840 kJ/kg) than that of gasoline (305 kJ/kg) ensures that
the volumetric efficiency of ethanol blend is higher than the efficiency
of pure gasoline, thereby improving power output [11].
Bioethanol is an eco-friendly oxygenated fuel as it contains 34.7%
oxygen, whereas, oxygen is absent in gasoline. This results in about
15% higher combustion efficiency of ethanol than that of gasoline [12],
thereby keeping down the emission of particulate and nitrogen oxides.
Compared to gasoline, ethanol contains negligible amount of sulfur,
and mixing of these two fuels helps to decrease sulfur content in the
fuel as well as emission of sulfur oxide, which is a carcinogen and can
contribute to acid rain [13]. Bioethanol is also a safer substitute to
methyl tertiary butyl ether (MTBE), which is commonly used as an
octane enhancer for gasoline and is added to the latter for its clean
combustion so that production of carbon monoxide (CO) and carbon
dioxide (CO2) can be reduced [14]. MTBE has been reported to make
its way into ground water that contaminates drinking water causing
severe detrimental effect on health [15]. The US Energy Policy Act
released an ANPR (Advance Notice of Proposed Rulemaking) in 2000
under the TSCA (Toxic Substance Control Act) to limit the use of MTBE
as a gasoline extender [16].
The renewable sources used for generating bioethanol can be
classified largely into sugars, starch, lignocellulosic biomass and algae.
Ethanol obtained from sugars and starch is referred to as the first
generation bioethanol, while lignocellulosic biomass and algae produce
second and third generation bioethanol, respectively. Production of
third generation bioethanol from algae is still in an immature stage and
confined to the laboratory research, while other types of biomass have
shown potential as bioethanol feedstocks on commercial scale. On the
other hand, conversion of three major types of feedstocks (sugars,
starch and lignocellulosic biomass) into bioethanol differs significantly,
particularly with regard to the obtainment of sugar solutions. Sugar
based raw materials require only an extraction process to get fermen-
table sugars, while starchy crops need to undergo hydrolysis to convert
starch into glucose. Lignocellulosic biomass has to be pretreated before
hydrolysis in order to alter cellulose structures for enzyme accessibility.
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Renewable and Sustainable Energy Reviews 71 (2017) 475–501
A comprehensive review on different raw materials, technological
approaches and process parameters are expected to play significant
roles on the overall research of bioethanol production. Taking into
account the above facts, this review paper has been designed to discuss
(1) potentials, composition and ethanol production process for differ-
ent renewable sources (2) technological achievements, (3) role of
microorganisms, and (4) effects of different factors on the process.
The content of the paper has been confined to the 1st and 2nd
generation ethanol.
2. Biomass sources for bioethanol production
2.1. Sugar sources
Sugar based raw materials include some energy crops, such as
sugarcane, sugar beet and sweet sorghum; fruits like grape, dates,
water melon and apple; and the sugar refinery wastes, namely cane
molasses and beet molasses [17]. The major advantages of sugar crops
as ethanol feedstocks are high sugar yield and low conversion costs,
while the seasonal availability of these crops is the main limiting factor
[1]. With regard to the ethanol production, each sugar crop or its
industrial wastes have some potential as ethanol feedstock that are
summarized in Table 1.
Sugarcane is a C4 plant having high capability to convert solar
radiation into biomass [19], and grown in tropical and sub-tropical
countries. The stalk juice of sugarcane and by-product of sugar
refineries (molasses) has been used as promising feedstocks for
bioethanol production over many years. Sugarcane juice is the main
feedstock in Brazil that produces about 79% of its bioethanol from this
feedstock, whereas molasses is the major ethanol feedstock in India
[22,23]. Cane juice has sufficient organic nutrients and minerals, in
addition to containing readily fermentable sugars that have made it an
ideal raw material for bioethanol. However, sugar concentration in
juice varies considerably depending on the variety, maturity and
harvest time.
Sweet sorghum is another C4 plant and a potential energy crop for
its unique characteristics, including high carbon assimilation (50 g/
m2/day), efficiency to use high water, and ability to accumulate high
level of extractable sugars in the stalks [24]. Unlike sugarcane, it can be
cultivated in almost all temperate and tropical climate areas and can be
developed from seeds [25].
Sugar beet is a major source of sugars in Europe and North
America, and used for bioethanol production in France [1]. It has also
been successfully introduced in India and currently there are trials
being conducted in other tropical countries such as China, Australia,
Kenya, South Africa, Brazil and the USA [26]. It grows in temperate
climate and require a lower rainfall than in sugarcane [1]. Sugar beet
usually produces a root with average weight of 0.5–2 kg, which
contains most of the sugars. The concentration of sugars in the sugar
beet root varies depending to the variety and growth conditions [26].
Molasses is a dark, viscous and sugar rich by-product of sugar
refinery industries. It includes two well-known by-products, namely
cane molasses and beet molasses, which are produced from sugarcane
and sugar beet in the respective refinery. Molasses are traditionally
used as a feed ingredient and binder. Currently it is used as an
attractive raw material for bioethanol production. The composition
and sugar contents in molasses can vary depending on the processes
used for sugar extraction as well as the composition of starting
materials [19].
A number of fruits have also been investigated for bioethanol
production in different parts of the world that considered mainly the
waste fruits in a specific region or production area [27,28]. Usually,
good amounts of fruits are discarded at harvest and during marketing
due to a low quality or unacceptable physical appearance that make
these fruits unfit for human consumption. Fruits are enriched with
soluble sugars that can be easily fermented by yeast without any
H. Zabed et al. Renewable and Sustainable Energy Reviews 71 (2017) 475–501

Table 1
Potential of some sugary biomasses as ethanol feedstocks.

Biomass Major influential features Reference

Sugarcane • High biomass yield [18,19]

(Saccharum officinarum) • High sucrose content
• High efficiency to accumulate solar energy
• Global production is 360 million ton/year
• Crop yield is 60–79.5 t/ha
• Residues are good source for generating electricity and 2nd generation bioethanol
Sweet sorghum • More drought tolerant crop than sugarcane [1]
(Sorghum bicolor) • Can grow in the arid land
• Stalk juice is a promising feedstock for bioethanol, while grains are used for food and starch based ethanol production
• Stalks contain significant amounts of sucrose, glucose and fructose
• Average syrup yield from stalk is 1900 L/ha
Sugar beet • Major source of sugars in Europe and North America [1,18]
(Beta vulgaris) • Crop yield is 79.1 t/ha
• Average syrup yield is 60–120 t/ha
• Average ethanol yield is 95 L/ton of stalk
Watermelon • Around 20% of waste watermelons during or after harvesting of crops can be used as bioethanol feedstock [20]
(Citrullus lanatus) • Juice can be used as a diluent for molasses that increases overall sugar content in the feedstock and reduce water consumption
Dates • AContains
good amounts of dates are wasted each year in the date producing countries, which can be used as ethanol feedstock [21]
(Phoenix dactylifera) • Long timehigh amounts of individual sugars (sucrose, glucose and fructose) as estimated for roughly 70% of total sugars
• A renownedstorage
Molasses • An industrialethanol feedstock enriched with sucrose, glucose and fructose [19]
• Valorization ofwaste and thus no debate of food versus fuel.
• Molasses yield istheapproximately
waste material
• 3–7 t/100 t of sugarcane

sophisticated pretreatment process. In some cases, it is easier to collect
juice from fruits compared to the extraction method used for collecting
sugarcane juice. However, it is not realistic to use fresh fruits for
bioethanol production instead of human consumption as it raises
criticism. Some fruit refinery wastes, such as pineapple and grape
pomaces can also be used for bioethanol production [29–31].
2.2. Starch sources
Starchy crops are widely used for bioethanol production due to their
availability across the world, ease of conversion, storage capability for a
long period and high ethanol yield. These raw materials include cereals
(60–80% starch), tubers and roots (60–90% starch), legumes (25–50%
starch), and green and immature fruits (As much as 70% starch) [32].
Corn, sorghum grains, wheat, cassava, potatoes and sweet potatoes are
some renowned sources of fuel ethanol [33]. Individual crops can vary
in the conversion efficiency and final ethanol yield (Table 2).
Corn is a major cereal crop widely used for bioethanol production
on commercial scale and its usage has been increased dramatically in
recent years. Considering the geographic locations, North America
produces the largest amounts of corn, followed by Asia, Europe and
South America [34]. United States is the dominant producer of corn
ethanol, which produced a record amount of ethanol from this feed-
stock (14.3 billion gallons) in 2014, and at the same time it exported
roughly 825 million gallons of ethanol to 51 countries across the world
[48]. The USA has also made its goal to produce 36 billion gallons of
ethanol by 2022 using corn and corn stover [49]. Recently, certain
European countries has shown interest to produce ethanol from corn.
Therefore, corn ethanol has gradually become a global biofuel with a
great economic importance.
Cassava and potato are two important tuber crops having potential
for bioethanol production. Cassava ranks as sixth food crop in the
developing countries, after rice, wheat, corn, potatoes and barley. This
crop is originated from South America but can grow in both tropical
and subtropical climates, and tolerant to semi-arid conditions with a
rainfall as low as 500 mm [50]. Another promising character of cassava
is the ability to grow in a wide range of soils, where fertility of soil is not
necessary and thereby leaving the fertile soils for other crops. The
global production of cassava has been estimated over 12.5 t/ha in the
cropping year 2011 [45]. Potato is a seasonal crop grown in temperate
climatic conditions, but primarily it is available abundantly in the
northern hemisphere. It is one of the fourth important food crops
round the world. China is the dominant producer of potato that
produces around 20% of the total world potatoes. Among the other
countries, Russia produces 12%, India 8% and USA 8% of potato [45].
Sweet potato is a root crop grown in tropical and sub-tropical
regions as a perennial crop, whereas it is an annual crop in the
temperate regions [51]. It is native to Central and South Americas.
However, currently China is the leading producer of sweet potatoes that
produces roughly 80% of the world's production, while Uganda and
Nigeria produce roughly 3% of the global crop and rank as second and
third, respectively [42]. Globally, sweet potato is the seventh most food
crop after rice, wheat, corn, potato, barley and cassava, and the second
most important root and tuber crop after potato [52]. Sweet potatoes
have several cultivars that vary in color from white to orange and even
purple depending on the starch content. Generally, white fleshed types
are larger in size and contains higher amounts of starch (25–40%) than
other varieties [45]. For this reason, white varieties are less sweet and
not considered as a food source crop. Thus, these sweet potato varieties
can be considered for bioethanol production.
2.3. Lignocellulosic biomass
Lignocellulosic biomass can be divided into several groups, such as
energy crops (perennial grasses and other dedicated energy crops),
aquatic plants (water hyacinth), forest materials (softwood, hard wood,
sawdust, pruning and bark thinning residues), agricultural residues
(cereal straws, stovers and bagasse), and organic portion of municipal
solid wastes. Each group of biomass has some potential to be used for
ethanol production as summarized in Table 3. In general, lignocellu-
losic biomass appear to be the largest, promising, and abundant
throughout the world, which can be used to produce ethanol without
necessitating any extra land or interference on food and feed crop
production. The global production of plant biomass is roughly
200×109 t/year, where nearly 8×109-20×109 t can be used for biofuel
production [53,54].
Most of the agricultural residues come from four major crops,
which are corn, wheat, rice and sugarcane, while the rest of residues
contributes only a little amount to the total world biomass [53]. Corn
stover is one of the most promising agro-residues having much

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H. Zabed et al. Renewable and Sustainable Energy Reviews 71 (2017) 475–501

Table 2
Potential of some starchy crops as bioethanol feedstocks.

Crops Major influential features Reference

Corn • Corn ethanol is a mature technology [18,34,35]

(Zea mays) • Not only the main feed stream but also the wasted corn contributes to the overall supply of raw materials.
• capability 9
Around 5% of the corn is wasted each year that can produce about 9.3×10 L of ethanol with a replacement
9
of about 6.7×10 L of gasoline.
• Protein rich co-product of corn ethanol plant is used as animal feed
• High fermentable hybrids can easily be developed for improved ethanol yield
Wheat • Conversion efficiency of wheat starch into ethanol is around 95% [18,36]
(Triticum aestivum) • The annual gross energy production for wheat derived bioethanol is 66 GJ/ha
• Kernel yield is 5.1 t/ha
Cassava • ACantropical root crop produced by many countries [18,37]
(Manihot esculenta) • Cassavabe easily hydrolyzed by various techniques
• with a lowerstarch does not have much application in food industries compared to corn starch, thereby available
price
• Available throughout the year due to its flexibility in terms of planting and harvesting
• Crop yield is 13.6 t/ha
Barley • About two billion gallons of ethanol can be produced per year from barley in North America [34,37]
(Hordeum vulgare) • Yield may vary between 0.82 t/ha and 3.08 t/ha with a global average of 2.5 t/ha
• Can be grown in areas that normally not used for corn, and hence, a barley-based ethanol industry will benefit
farmers and rural economy outside the ‘‘corn belt’
• Winter barley can be double-cropped with corn and soybean to give farmers three crops in each two-year cycle
• Winter barley as a cover crop can prevent loss of nitrates, phosphates, and sediments into watersheds and hence
provide protection for the environment
Canna • ACannon-food biomass source [38,39]
(Canna edulis) • Yearlybeoutput
cultivated in marginal lands and in subtropical highlands with low nutrient demand
• Enrich with starchis approximately 4.5 kg/m 2

Sorghum (grain) • Ability to grow in alike corn [40]

(Sorghum bicolor) • Efficient in water usage wide range of soil types and climates
• Drought tolerant
• An important staple crop in terms of total biomass
Sweet potato • Produced globally [41,42]
(Ipomoea batatas) • Drought resistant crop requiring low chemical and fertilizer inputs
• Can be grown in marginal soils
• Cheap substrate and rich in starch
Potato • Processing is easier than that of other grains [43,44]
(Solanum tuberosum) • Around 5–20% of the crops are wasted that can be used for bioethanol production
• Global production is more than 140 million ton/year
• Large starchy tuber and produced both annually and perennially in Africa, America, Caribbean, South Africa and
Yam
(Dioscorea rotundata)
• Asia [45]

• Several principal species are widely grown throughout the tropics, which are white yam (D. rotundata), yellow
yam (D. cayenensis), bitter yam (D. dumetorum) and water yam (D. alata)
Jerusalem artichoke (Helianthus tuberosus) • ATuber
tuberous-rooted perennial crop [45]
• Inulin iscontent
rich in synanthrin and other fructose polymers
• Sugars are storedin fresh tuber is about 10–20% with an average of 15%
• High alcohol potential in the roots and tubers
• No requirement of fertile soil for its growth
• Next season's crop is produced from small tubers left in the field, so no ploughing or seeding is necessary
• Can be used as basic material for bioethanol production
Iles-iles • Non-food ethanol feedstock [46]
(Amorphophalus campanulatus) • High carbohydrate content
• Low price
• Global oat production is 2.67×10 t/year
Oat • Yield can vary from 1.54 to 2.31 t/ha with an average yield of 1.98 t/ha
7
[34]
(Avena sativa) • Utilization of only wasted oat grain could produce about 225 million liter of ethanol that can replace 161 million
• liter of gasoline
• Dry milling can produce 1.5 kg distiller's dried grains with solubles (DDGS) per kg of ethanol that can replace
usage of oat as animal feed
Banana • Fruit, pulp and skin are enriched with starch [47]
(Musa sp.) • Valorization of waste if banana peel is used for ethanol production
• Ethanol production from banana has shown a positive energy balance
potential for bioethanol. It is a left over residue after harvesting the bagasse are generated in north and South America, respectively.
corn grains and consists of stalks, leaves, cobs and husks. An average Biomass based ethanol industry requires a continuous and reliable
annual production rate of corn stover is 4 t/acre [34]. Wheat straw is supply of raw materials to maintain a low cost ethanol production.
the residue of harvested wheat producing 1–3 t/acre annually. Rice Energy crops hold potential in this regard, which require short growth
straw is one of the most abundant and promising biomass in the world period, and minimal use of water, fertilizer and cultivable lands. These
that includes stems, leaf blades and leaf sheaths, with a global crops are mostly dedicated for biofuels and may be either C3 or C4
production rate 731 million ton/year [53]. Bagasse is a cheap residue plants. Some potential energy crops are miscanthus (Miscanthus spp.,
derived from the processing of sugarcane, which has been estimated to C4), switchgrass (Panicum virgatum, C4), reed canary grass (Phalaris
317–380 million ton/year [70]. Most of the rice and wheat straws are arundinacea, C3), giant reed (Arundo donax, C3) and alfalfa (Medicago
produced in Asian countries, while majority of the corn stovers and sativa, C3). In comparison to other sources (such as woods and C3

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H. Zabed et al. Renewable and Sustainable Energy Reviews 71 (2017) 475–501

Table 3
Potential of lignocellulosic biomass as ethanol feedstocks.

Biomass Major influential features Reference

Perennial grasses • High biomass yield [55–57]

Aquatic plants
Agricultural residues
Forest biomass and waste
Municipal solid waste (MSW)
• High cellulose content
• Easy to grow and harvest
• Having potential to cover 50–70% of the total bioethanol feedstocks
• Yield can be ranged from 0.9 to 37 t dry matter/ha
• Potential ethanol yield is 160–460 L/ton of biomass
• Grow in water bodies without competing with arable lands [58,59]
• Exceptionally fast growing plant
• Can also be used for water purification to extract nutrients and heavy metals
• Biomass productivity is very high
• Widely prevalent aquatic weeds
• Abundant in certain parts of the world that has made it a suitable feedstock for distributed ethanol production
• Easily available [34,60–62]
• Valorization and eco-friendly management of agro-wastes
• Minimize the reliance on forest woody biomass and reduce deforestation
• Short harvest rotation period
• Global crop residues have been estimated to 2802 million ton/year for total cereal crops, 3107 million ton/year for 17 cereals
and legumes, and 3758 million ton/year for 27 food crops
• Potential ethanol yield is 235–450 L/ton of biomass
• AHigh
lower ash content compared to crop residue [63–66]
• Flexibledensity of the biomass that offers more economical transportation
• One of theharvesting time
• Potential ethanol
largest unexploited and underutilized biomass
• Valorization of MSW yields are 220–275 L/ton for softwoods and 280–285 L/ton for hardwoods
• An estimated abundance to fuel ethanol that can correlate both energy security and waste management [67–69]
• Potential ethanol yield is of154MSW is 20.7 million ton/year for an urban area with 217 million of population
• L/ton of biomass

crops), C4 perennial grasses produce more than two times biomass
each year in warm and temperate regions due to their more efficient
photosynthetic pathway [55].
Forest biomass include mainly woody materials such as hardwoods
and softwoods, while forest wastes are sawdust, pruning, bark thinning
residues, wood chips and branches from dead trees. Woody biomass
has been estimated roughly 370 million tons per year in the USA [71].
Other countries rich in forests are Canada, Russia, Brazil and China
that comprise together more than half of the total forest area of the
world [72]. Softwoods is derived from the evergreen species such as
pine, cedar, spruce, cypress, fir, hemlock, and redwood, which possess
low densities and fast growth. Hardwoods are mainly found in the
Northern hemisphere and include trees such as poplar, willow, oak,
cottonwood and aspen.
Municipal solid wastes (MSW) is a recyclable biomass that originate
from residential and non–residential sources such as food wastes and
paper mill sludge [60]. The estimated global production of MSW was
1.3×109 ton in 1990, which became almost double after 10 years,
accounting for 2.3×109 ton in 2000 [72]. Mostly the organic fraction of
MSW is used as ethanol feedstock. However, MSWs are rarely
considered as ideal feedstocks for bioethanol due to their wide
diversification in composition and existence of microbial contamina-
tion, even though these feedstocks have the potential in regional
perspectives.
most competitive over other two feedstocks as it reduces the green-
house gas emissions by 30% and 40% compared to corn and sugar beet,
respectively. The production costs of sugarcane based ethanol is
significantly lower than the corn by 60% and sugar beet by 75%. The
potential ethanol yield for sugarcane was reported to be the higher as
estimated 6470 L/ha for sugarcane, 4180 L/ha for corn and 5500 L/ha
for sugar beet [74].
The potential of different lignocellulosic biomass for ethanol
production is primarily determined by the efficiency of glucan hydro-
lysis into soluble sugars. In an earlier study, a great variation was
reported in glucan digestibility among some non-pretreated biomass
including agricultural wastes, bioenergy crops and woody biomass [75].
Among the agricultural residues, the order of glucan digestibility was
reported as bagasse (47%) > corn stover (23%) > hurd of industrial
hemp (14%) > wheat straw (11%) > rice straw (10%). On the other
hand, bioenergy crops showed a lower digestibility than that of
agricultural residues and gave an order of common reed (19%) >
switchgrass (17%) > Miscanthus (8%) > bamboo (3%). The non-
pretreated woody biomass such as poplar showed a glucan digestibility
of ~7% [75]. Table 4 compares some well-known biomass sources
based on the ethanol yield reported in certain literature.
3. Carbohydrate content in different biomass

Carbohydrate is the main fermentable components in a bioethanol

2.4. Ethanol yield from different biomass: a comparative scenario
Although numerous biomass sources have been used for generating
bioethanol either on commercial or laboratory scale, sugar feedstocks,
particularly sugarcane, provides cheaper ethanol compared to starch
and lignocellulosic biomass based feedstocks [73]. However, the final
ethanol yield of a biomass source depends on the overall conversion
efficiency, which further varies with regard to the nature of biomass,
process conditions and microorganisms used in fermentation. In an
earlier study, a comparative study was carried out using three major 1st
generation ethanol feedstocks used in Brazil (sugarcane), USA (corn)
and Europe (sugar beet) [74]. According to this study, sugarcane is the
feedstock, which include, in general, soluble sugars (mono and
disaccharides), starch, cellulose and hemicellulose. Soluble sugars are
the primary components in sugar based feedstocks (Table 5), while
starch is the major carbohydrate in the starchy crops (Table 6).
Cellulose and hemicellulose are the principal carbohydrates in ligno-
cellulosic biomass (Table 7). Carbohydrate contents in renewable
sources can vary significantly depending on the type of biomass,
botanical sources and crop hybrids. The carbohydrate containing parts
in crops that are used as ethanol feedstocks also differs among the
crops, and include stalks, grains, tubers, roots, leaves, stems, fruits, and
straws.
Fermentable sugars present in the biomass, particularly sugar

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Table 4 Table 6
Comparison of different biomass with regard to the ethanol yield. Starch content in some starchy crops.

Biomass Ethanol yield (L/ Potential yield (L/ Reference Crops Scientific name Crop Starch Reference
ton) ha) type content
(%)
Sugarcane 70–90 6470–6660 [74,76,77]
Sugar beet 95–107 1605–5500 [74,76,77] Corn Zea mays Cereal 70–72a [94]
Corn 370–470 4180 [74,77,78] Sorghum Sorghum bicolor Cereal 68–70.7a [40]
Sorghum stalk 40–86 2062–2595 [77–79] Wheat Triticum aestivum Cereal 65.3–76 [36,95]
juice Rice Oryza sativa Cereal 87.5a [34]
Sorghum bagasse 250 1796–6591 [78,80] Oat Avena sativa Cereal 65.6a [34]
Sorghum grain 380 1099 [76,78] Potato Solanum tuberosum Tuber 17.5–20b [45,96]
Molasses 280 – [77] 73a
Barley 345 – [81] Sweet potato Ipomoea batatas Root 14–27. 5b [97,98]
Oat 264 – [81] Sugar beet Beta vulgaris Root 8–12b [45]
Triticale 368 – [81] Barley Hordeum vulgare Cereal 62.7–68.7a [99,100]
Sweet potatoes 125–170 1989–4800 [76,77,82] Peas Pisum sativum Legume 46.2a [101]
Cassava 363–455 4901 [76] Cassava Manihot esculenta Tuber 35b [102]
Jerusalem 90 1821–5930 [62,81] Canna Canna edulis Tuber 11–15.6b [39]
artichoke Banana Musa paradisiaca Fruit 27.24b [103]
Arrowroot 208 2496 [76] Yam Dioscorea rotundata Tuber 20–40b [104]
Potato 80–100 1600 [76,77] Taro (Aroids) Colocasia esculenta Root 15–25b [104]
Wheat 376–435 1001–1700 [76,81] Jerusalem Helianthus tuberosus Tuber 15b [105]
Yam 192 4800 [76] artichoke
Corn stover 450 4400 [62,83] Iles-iles Amorphophalus Tuber 79a [46]
Corn cob 510 – [62] campanulatus
Wheat straw 490 – [62] Babassu Orbygnia phalerata Palm 60.05a [106]
Breadfruit Artocarpus altilis Fruit 64.5a [107]
Buckwheat Fagopyrum Pseudo 80.5a [108]
Table 5 tataricum cereal
Sugar content in some sugar based ethanol feedstocks. Chestnut Castanea sativa Nut 93.2a [109]
Ginger Zingiber officinale Rhizome 85a [110]
Crops Feedstock Total soluble Reference Kudzu Pueraria lobata Root 99.5a [111]
sugars (%) Lentil Lens culinaris Legume 51.7a [112]
Millet Panicum sumatrense Cereal 70a [113]
Sugarcane Stalk juice 12–17.6a [73,84] Peruvian Arracacia Root 80a [114]
Molasses 46b [19] carrot xanthorrhiza
Horse gram Dolichos biflorus Legume 60.5a [115]
Sweet sorghum Stalk juice 16–21.8 a
[85,86] flour
Sugar beet Root juice 15–20a [26]
a
Molasses 52.9b [87] Dry weight;
b
Fresh weight.
a
Watermelon Fruit juice 7–10 [20]
White grape (Vitis Pomace 14.8–49.1b [88,89] the amount of starch [124,125]. Starch is composed of two polymers of
vinifera)
b α-D-glucose units, namely amylose and amylopectin. Amylose is linked
Apple (Malus Pomace 21.4–31.8 [90]
domestica) by α-1→4 glyosidic bonds, while both α-1→4 and α-1→6 linkages are
Dates Date industry 73–83 b
[28] found in amylopectin. Amylose is a linear polymer of around 1000
waste glucose units, while amylopectin possess a highly branched configura-
b
Juice 44–88 [91]
tion along with a branch that linked by α-1→6 bonds in every 20
Mahula (Madhuca Flower 36–38a [92]
linkages [126]. The physicochemical properties of starch are often
latifolia) associated with its amylose and amylopectin ratio, which significantly
– Cheese whey 4.5–5a* [93] affect final ethanol yield [124,127].
Lignocellulosic biomass contain varying amounts of cellulose,
a
Fresh weight;
b
hemicellulose and lignin depending on the types of biomass
Dry weight,
*
Main sugar is lactose. (Table 7). Cellulose and hemicelluloses constitute roughly two–third
of the total dry weight of the biomass that are the main substrates for
based raw materials are mainly glucose, fructose and sucrose. These ethanol [128]. In the native biomass, cellulose and hemicellulose
soluble sugars can be fermented and converted into ethanol by yeasts polymers remain connected with each other by the covalent and
or other fermenting microorganisms without requiring any further hydrogen bonds and both further linked to lignin, thereby forming a
treatment. Unlike soluble sugars, starch is a polymer of glucose that complex lignocellylosic matrix, which is highly robust and recalcitrant
cannot be metabolized by the traditional yeasts and bacteria in its to depolymerization [129]. In general, recalcitrance of biomass is
polymeric stage. Consequently, starch must be converted into glucose associated with the properties of biomass, including the crystalline
prior to subjecting it into fermentation, which requires amylolytic nature of cellulose, low available surface area, protection of cellulose
enzymes and is known as hydrolysis. and hemicellulose by lignin, and heterogeneous nature of the biomass
Starch is the major storage carbohydrate in plants and it acts as particles. The composition and structure of the biomass determine the
energy reservoir for higher plants, such as cereals, legumes and tubers digestibility of lignocellulose to subsequent chemical or biochemical
[123]. However, the bioavailability of starch during enzymatic hydro- treatments [130,131].
lysis depends on its structure that can vary with regard to the botanical Cellulose is a linear and crystalline structure containing linear
sources and crop hybrids. This variation in starch structure signifi- chains of glucose units and joined together by β–1→4–glucosidic
cantly affects the overall conversion efficiency, sugar yield as well as linkage, with and average molecular weight of about 100,000 Da
final ethanol yield. For this reason, starch conversion efficiency and [118]. Extensive hydrogen bond within the cellulose molecules create
ethanol yield have not been reported to be significantly correlated with its crystalline network through cross linking of various hydroxyl

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Table 7
Holocellulose and lignin contents in some lignocellulosic biomass.

Biomass Cellulose (%) Hemicellulose (%) lignin (%) Reference

Switchgrass 5–20 30–50 10–40 [116]

Miscanthus 38–40 18–24 24–25 [117]
General grasses 25–40 25–50 10–30 [53]
Municipal solid waste 33–49 9–16 10–14 [69,118]
Newspapers 40–55 25–40 18–30 [119]
Corn cob 42–45 35–39 14–15 [54,68]
Corn stover 38–40 24–26 7–19 [53,120]
Sugarcane bagasse 42–48 19–25 20–42 [53,121]
Rice straw 28–36 23–28 12–14 [53]
Wheat straw 33–38 26–32 17–19 [53]
Barley straw 31–45 27–38 14–19 [53]
Sweet sorghum bagasse 34–45 18–27 14–21 [53]
Oat straw 31–37 27–38 16–19 [70]
Rye straw 33–35 27–30 16–19 [70]
Rice husk 25–35 18–21 26–31 [122]
Softwood 27–30 35–40 25–30 [116]
Hardwood 20–25 45–50 20–25 [116]

groups, and finally produce micro fibrils that make cellulose molecules
more strength and rigid [60]. This explicit structure of cellulose results
in its tightly packed backbone and high crystallinity, which in turn,
causes insolubility of the cellulose polymer to water and resistance to
depolymerization [131].
Hemicellulose is a complex and heteropolymer with an average
molecular weight of nearly 30,000 Da. It consists of short, linear and
highly branched chains of different monomers including hexoses (β–
D–glucose, α–D–galactose and β–D–mannose), pentoses (β–D–xylose
and α–L–arabinose). The hemicellulose chains may also contain sugar
acids (uronic acids) namely, α–D–glucuronic, α–D–galacturonic and
α–D–4–O–methylgalacturonic acid, which remains associated with
cellulose in the plant cell wall [60,118]. Sometimes other sugars such
as a small amount of α–L–rhamnose and α–L–fructose may present in
hemicellulose. The backbone chain of hemicellulose is principally made
up of xylan through β–1, 4 linkages that include D–xylose (around
90%) and L–arabinose (roughly 10%) [128]. The commonly known
hemicelluloses are xylans and glucomannans. Xylans are the main
hemicellulose in hardwoods, forest wastes, agricultural residues, and
municipal and industrial wastes, while softwood are typically rich in
glucomannan [60,128].
Lignin is a highly branched aromatic polymer present in the plant
cell wall, and is linked to cellulose polymers forming a lignocellulosic
matrix [132]. The monomers of a lignin polymer are three phenolic
compounds such as coumaryl, coniferyl and sinapyl alcohol. Among the
lignocellulosic biomass, woods contain relatively higher amounts of
lignin, where softwoods range from 30% to 60% and hardwoods vary
between 30 and 55% of lignin. Grasses and agricultural residues
contain low amounts of lignin, ranging from 10% to 30% and 3–
15%, respectively [60].
4. Bioethanol production from sugary biomass
Ethanol production from sugar containing raw materials is a well-
known and mastered process. Sugar based ethanol is often referred to
as the Brazilian fuel as it produces the second largest ethanol in the
world from sugarcane, after the US corn ethanol. To date, many
investigations have been carried out using different sugar based feed-
stocks in various technological aspects that have given a general
scenario for ethanol production from these raw materials (Table 8).
In a general procedure illustrated in Fig. 2, juice is extracted from
the sugar crops by crushing stalks (sugarcane and sweet sorghum) or
root (sugar beet) in a specialized roller. The collected cane juice is then
purified by lime. Any suitable source of lime can be used, but lime milk
[Calcium Hydroxide; Ca(OH)2] or calcium saccharate are preferred.
The addition of the source of lime raises the pH of the sugar cane juice.
Lime is added up to a maximum concentration of about 2% by weight
of the solids content of the juice. The addition of lime in the juice
reduces its colorants, neutralizes organic acids contents, and forms
calcium phosphate precipitate, which can be removed from the juice
through sedimentation [138].
Filtration is applied after lime purification to remove debris from
the juice, and the removed fraction is often known as filter cake. The
filtrate sugar solution is evaporated to condense it to a sugar level
ranged between 14% and 18%, considering the sugar tolerance capacity
of the fermenting microorganism [1]. The concentrated syrup is
supplemented with ammonium sulfate or other nitrogen sources,
sterilized, pH and sugar concentration adjusted and then fermented
by microorganisms, preferably yeast.
The co-product and residues obtained from a typical sugar based
ethanol plant can be used for different purposes. There are two
residues, namely bagasse and filter cake, produced during extraction
and purification of juices. Bagasse is used to generate electricity for the
plant that meets the energy demand and reduce the production costs,
while filter cake is used as an eco-friendly fertilizer in the agricultural
field. Sugarcane bagasse is also an attractive raw material for ligno-
cellulosic ethanol production. The co-product, vinasse, is an important
animal feed, which can also be used as fertilizer.
5. Bioethanol production from starchy biomass
As was discussed above, sugar crops require only an extraction
process to get fermentable sugars, while starchy crops need to undergo
a hydrolysis step to convert starch into glucose that uses preferably
amylolytic enzymes. As a result, it is not a matter of surprising that
sugar crops produce cheaper ethanol than the starchy crops. However,
sugar crops cannot be grown globally due requirement of selective
climatic conditions and soil types [140]. Nonethless, starch based
bioethanol is relatively well established and produces about 60% of the
total ethanol, compared to nearly 40% as produced from sugar sources
[141,142].
Currently, ethanol is produced from corn as well as other starchy
crops either by the dry grind or wet mill method, obtaining 9.5 L/
bushel ethanol in the latter to 10.6 L/bushel in the former using corn as
feedstock [94]. The basic difference between dry-grind and wet mill
methods is the usage of whole ground grains or tubers in the former,
while different components are first separated from the raw materials
and only starch is used in the wet mill method (Figs. 3 and 4). Different
co-products are produced in either method that contributes to the
overall economy of the process. Although both processes are now being
employed in ethanol production, majority of the commercial plants use
dry-grind method. The wet mill method requires extensive equipment

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Table 8
Recent investigations on the bioethanol production from sugar crops and sugar based agro-industrial wastes.

Feedstock Objectives Major findings Reference

Sugarcane juice • Evaluation of the mineral composition in juice of three • The content of some important minerals, such as Cu, Mg, Zn and
sugarcane verities P were mentionable that varied with sugarcane verities
[133]

• Studying the effect of mineral content in juice on the • Maximum amount of ethanol was obtained from the highest
fermentation efficiency of S. cerevisiae Mg containing juice
• Studying the effects of yeast cells adaptation to galactose on the • A 30% higher amount of ethanol was produced by the galactose
final ethanol yield during fermentation by a thermotolerant adapted cells than that of non-adapted cells.
[84]

yeast strain (Pichiakudriavzevii kudriavzevii), isolated from

sugarcane juice through enrichment technique
• Fermentation with galactose adapted cells at 40 °C produced an
ethanol concentration of 71.9 g/L and productivity of 4.0 g/L/h
Sugar beet juice • Studying the effects of supplementation of juice with mineral • A high concentration of ethanol was produced by the
salts on ethanol yield supplemented juice that ranged between 85 g/L and 87 g/L
[134]

Sweet sorghum • Evaluation of ethanol yield of five sweet sorghum varieties, • Ethanol concentration in the broth varied significantly among
namely Keller, BJ 248, SSV84, Wray and NSSH 104 the verities, where the Keller variety produced maximum
[135]

amount of ethanol (9%, w/v)

Watermelon juice • Investigation of watermelon juice as a diluent, additive and • Complete watermelon juice that did not contain lycopene but
nitrogen source during sugar or molasses fermentation contained free amino acids, were readily fermentable either as
[20]

main feedstock or as diluent, supplement, and nitrogen source

to granulated sugar or molasses. A minimum level of ~15 μmol/
ml amino nitrogen was required in the juice to achieve
maximum fermentation rates when it was employed as the sole
nitrogen source for the fermentation.
• Studying
yield
the effect of pH on fermentation efficiency and ethanol • As much as 25% sugar was fermented at pH 3 with an ethanol
concentration range of 0.41–0.46 g/g
• Up to 35% sugar was fermented at pH 5 with an ethanol
concentration range of 0.36–0.41 g/g

Dates • Comparison between direct and soxhlet extraction of sugars • Although direct extraction method provided slightly
from three date verities (Kunta, Eguoua and Bouhatem) concentration of sugars (104.6–214 g/L) than soxhlet
lower [21]

extraction (115.1–225.8 g/L), it required significantly lower

time and energy
• Study of the date verities for their bioethanol potential • Ethanol produced by the varieties was very close, around 25%
(v/v)

Yam bean
(Pachyrhizus
• Fermentation
fermentation
of yam bean juice with batch and repeated batch • Maximum ethanol was obtained from the repeated
fermentation that showed the highest efficiency
batch [136]

erosus)
Sugar beet molasses • yield
Studying the effects of using diluted molasses broth on ethanol • Dilution of the molasses negatively correlated with ethanol yield,
and undiluted broth produced maximum amounts of ethanol
[26]

• Studying the effects of pH and yeast inoculum concentration on • Low pH (4) and moderate inoculum size (5 g/L) produced
ethanol yield maximum amounts of ethanol
• Studying the effects of supplementation of media with nitrogen • Supplementation with ammonium sulfate significantly
source on ethanol yield enhanced final ethanol yield

Sugarcane molasses • Evaluation of the performance of three reactors and dilution • The MBR worked with high substrate concentration and found
rates of cane molasses during continuous ethanol fermentation promising for scale up of the operation
[137]

with regard to ethanol productivity. The reactors were

continuously stirred tank reactor (CSTR), immobilized cell
• The CSTR showed minimum productivity (6.8 g/L/h), while
MBR produced maximum amount of ethanol (46.5 g/L) with a
reactor (ICR) and membrane reactor (MBR); productivity of 19.2 g/L/h
Whey • Investigation of bioethanol production from whey
fermenter integrated with direct contact membrane
in a • Afromcontinuous removal of ethanol and other volatile compounds
the broth by membrane distillation resulted in a high
[93]

distillation (MDBR) efficiency of sugar conversion into ethanol.

• Fermentation rate is significantly faster in the integrated
bioreactor than in the traditional rector
• The concentrated whey and de-proteinized whey enriched with
either sucrose or lactose were used as fermentation media
• The efficiency of ethanol production by the deproteinized whey
enriched with sucrose was close to the theoretical value

Mahula flower
(Madhuca
• Comparison of efficiency and yield during fermentation of • S. cerevisiae showed a higher fermentation efficiency than Z.
mahula flower juice by two widely used microorganisms (S. mobilis, with 21.2% higher ethanol yield, 5.27% higher
[92]

longifolia L.) cerevisiae and Z. mobilis) productivity and 134% higher sugar conversion by S. cerevisiae

and high capital investments, producing large amounts of ethanol and
a variety of co-products. On the other hand, dry-grind process is
suitable for generating ethanol on a small scale requiring less equip-
ment and capital investment that produces two major products such as
ethanol and distiller's dried grains with solubles (DDGS). Furthermore,
a higher ethanol yield has been reported in the dry grind ethanol
production from corn (0.395 L/kg) than that of wet mill method
(0.372 L/kg) [143]. As a result, dry-grind process is the most attractive
and widely used for generating bioethanol. Moreover, a recent growth
in the corn ethanol industry in the USA has been made mainly on dry-
grind plants, which produces 70–86% of total ethanol [144–146].
The conventional dry-grind method involves preparation of slurry
by mixing corn flour with water, which is then cooked and liquefied
with a thermostable α-amylase to breakdown the starch molecules into
dextrin and other smaller molecules. The liquefied slurry is then
saccharified to convert dextrin into glucose using glucoamylase, and
subsequently subjected to yeast fermentation to produce ethanol from
glucose. After fermentation, ethanol is recovered through distillation of
the beer, leaving whole stillage, which is then separated into thin
stillage and solid fraction. One portion of the thin stillage is recycled as
backset in the process to decrease water consumption and another
portion is condensed into thick stillage (syrup). The solid portion is
mixed with thick stillage to obtain a final co-product, DDGS, which is
sold and used as animal feed [149]. In different recent investigations

482
H. Zabed et al.
Fig. 2. Schematic diagram of ethanol production from sugar biomass [139].
with starchy materials, much efforts have been made to improve overall
conversion process and ethanol yield using feedstocks from various
botanical sources (Table 9).
6. Bioethanol production from lignocellulosic biomass
The general procedure for producing lignocellulosic ethanol starts
from grinding of the biomass to reduce its particle size. The ground
biomass is then mixed with water to make slurry with an initial dry
solid load between 10% and 15% (Fig. 5). Generation of ethanol from
lignocellulosic biomass essentially needs to degrade lignocellulosic
network into its fractions that include individual cellulose, hemicellu-
lose and lignin polymers using a suitable pretreatment method. The
released cellulose and hemicellulose molecules present in the pre-
treated biomass are then hydrolyzed into soluble sugars via chemical or
enzymatic method, which are finally converted into ethanol during
microbial fermentation. Ethanol produced in the fermentation broth is
recovered and purified to obtain a fuel grade ethanol. The recovery
process is done using distillation method, which produces ethanol,
lignin residues and waste water. A portion of water is recalculated as
backset, while lignin can be combusted and converted into electricity
and heat. Therefore, the overall conversion process of lignocellulosic
can be divided into four major steps that include pretreatment,
Fig. 3. Schematic diagram for dry-grind ethanol production from starch (Adapted from [147]).
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Renewable and Sustainable Energy Reviews 71 (2017) 475–501
hydrolysis, fermentation and product recovery (Fig. 5).
Lignocellulosic biomass are versatile in their nature, particularly
physicochemical properties vary considerably with regard to the types
of biomass. These variations affect the conversion process and thereby
it is difficult to design a uniform conversion method and optimum
conditions for all biomass. Some recent investigations on the ethanol
production from lignocellulosic biomass are summarized in Table 10.
7. Pretreatment of the biomass
Pretreatment is the most complicated and costly step in the
conversion of biomass into ethanol. It alters macroscopic, sub–micro-
scopic and microscopic structures of the biomass. Pretreatment
primarily brings about the removal of lignin and hemicellulose from
the lignocellulosic matrix, decrease in crystallinity of cellulose, and
increase in the surface area and porosity of the biomass [176].
Pretreatment of the lignocellulosic biomass may also release some
fermentable sugars by depolymerizing the hemicellulose fraction of the
lignocellulose [177].
To date, various pretreatment methods have been investigated,
either simple or technologically and logistically more intensive, which
can be broadly classified into four categories, such as physical,
chemical, physicochemical and biological. These pretreatment methods
differ from each other with respect to the mode of action, reaction
conditions and overall outcomes. In particular, each pretreatment
technique has some advantages and disadvantages as shown in
Table 11.
Physical methods of pretreatment include mechanical grinding (ball
milling, two roll milling, hammer milling, colloid milling, and vibro
energy milling), thermolysis and irradiation (gamma rays, electron
beam or microwaves). In general, this type of pretreatment works on
the biomass by increasing the surface area and pore volume, decreasing
the degree of polymerization of cellulose and its crystallinity, hydrolysis
of hemicelluloses, and partial depolymerization of lignin [179].
Physical pretreatments are energy consuming, environmentally un-
friendly and non-viable for commercial process.
Lignocellulosic biomass can be pretreated chemically using primar-
ily alkali and acid that work on the biomass through delignification and
decreasing the crystallinity of cellulose. Various inorganic acids are
widely used during acid pretreatment, which include H2SO4, HCl,
H3PO4 and HNO3. In addition, some organic acids, such as fumaric and
maleic acids are also used for pretreatment of the biomass that produce
lower amounts of inhibitors than those produced during pretreatment
with inorganic acids. Acid can be used either at the dilute or
concentrate stages that offers some advantages and disadvantages over
each other (Table 11). The effective concentration for dilute acid
pretreatment is usually below 4%. It works at high temperature (e.g.
180 °C) during a short period of time; or at low temperature (e.g.
H. Zabed et al.
Fig. 4. Schematic diagram of wet mill ethanol production from starch (Adapted from [148]).
120 °C) for a longer retention time (30–90 min). Concentrated acid
pretreatment uses the concentration of the acid in the range between
70% and 77% at 40–100 °C. Among the alkali, NaOH, KOH, NH4OH
and Ca(OH)2 are commonly used for alkali pretreatment of biomass.
Alkali pretreatment can be performed at room temperature with a
reaction time ranging from seconds to days. Several cellulose solvents,
such as alkaline H2O2, ozone, glycerol, dioxane, phenol and ethylene
glycol are used for pretreatment, which disrupt cellulose structure and
promote hydrolysis of this carbohydrate polymer. Ionic liquids (ILs)
are other promising chemicals and typically composed of large organic
cations and small inorganic anions. ILs exist as liquids often at room
temperature and have a tendency to remain liquid in a wide range of
temperatures ( < 100 °C). Organosolv is a popular pretreatment meth-
od that use a mixture of organic or aqueous solvents such as methanol,
ethanol, acetone and ethylene glycol. The solvents are usually com-
bined with acid catalysts such as HCl, H2SO4, oxalic or salicylic to
break hemicellulose bonds.
Physicochemical pretreatment exploit its action by combining the
physical conditions and chemicals. A good number of technologies has
been introduced till to date under this category of pretreatment, which
include steam explosion, ammonia fiber explosion (AFEX), ammonia
recycling percolation (ARP), soaking aqueous ammonia (SAA), wet
oxidation, CO2 explosion and so on. Physicochemical methods exert the
actions on biomass through increasing the accessible surface area,
decreasing the crystallinity of cellulose, and removing hemicelluloses
and lignin from the lignocellulose.
Biological pretreatment of lignocellulosic biomass can be carried
out using microorganisms, particularly fungi, which are white rot,
brown rot and soft rot fungi. Among these pretreating microorganisms,
white rot fungi are the most efficient [180]. Brown rot fungi attacks
cellulose, while white and soft rot fungi attack both cellulose and lignin
[68]. The biological conversion of different lignocellulosic biomass,
such as forest and agricultural residues, and dedicated energy crops
offer some benefits over chemical and physical methods. The major
disadvantages of biological pretreatment are lower hydrolysis rate and
requirement of longer incubation time compared to other pretreatment
methods.
8. Formation and detoxification of inhibitors
The severe conditions applied during thermochemical pretreatment
often lead to partial breakdown of the major lignocellulosic compo-
nents (cellulose, hemicellulose, lignin, extractives and ash) into differ-
ent products, which are acetic acid, formic acid, levulinic acid,
hydroxymethyl furfural (HMF), furfural and other phenolic com-
pounds. These by-products act as inhibitors for both fermenting
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Renewable and Sustainable Energy Reviews 71 (2017) 475–501
microorganisms and cellulose degrading enzymes when their accumu-
lation is sufficiently high [181]. The inhibitors are categorized into
three major groups based on the origin such as aliphatic acids, furan
derivatives and phenolic compounds [182,183]. Formation of different
breakdown products are outlined in Fig. 6. The type and amounts of
inhibitors generated during pre-treatment depend on both the nature
of the biomass and pretreatment conditions, which include tempera-
ture, time, pressure, pH, redox conditions, and addition of catalysts
[184].
There are several ways to counteract the problems with inhibitors
by removing or neutralizing them from the pretreated solution. These
methods are broadly classified into physical, chemical, physicochem-
ical, biological and newly emerging techniques (Fig. 7) [185]. An
efficient detoxification method should selectively remove inhibitors,
and be cheap and easy to integrate into the process [182]. In general,
removal of inhibitors can be done by some renowned techniques that
include extraction, ion exchange, active coal, overliming, steam strip-
ping or evaporation at low pH, and enzymatic action using laccase and
peroxidase [184]. Stripping or evaporation is effective for removing the
volatile inhibitors (e.g., acetic acid and furan). Over-liming detoxifica-
tion of the solution include the addition of Ca(OH)2 up to a pH reached
to 11 that removes both volatile and non-volatile inhibitors such as
(e.g., furans and phenols) [184].
9. Hydrolysis
Hydrolysis is required for converting polymeric carbohydrates
(starch, cellulose and hemicellulose) into fermentable sugars, since
yeast cannot utilize carbohydrate polymers as a substrate [188]. In
general, hydrolysis of starch, hemicellulose and cellulose can be done
either by acidic or enzymatic methods, and the outcomes of hydrolysis
are usually dependent on the type and composition of the biomass and
hydrolysis conditions.
9.1. Hydrolysis of starch
During hydrolysis of starch, the amylose and amylopectin mole-
cules undergo biochemical conversion to produce soluble sugars.
Although acid hydrolysis is an effective way of generating fermentable
sugars from starch, the necessity for acid recovery after hydrolysis has
made it unattractive [189]. Moreover, acid hydrolysis is often asso-
ciated with the production of undesirable byproducts, most impor-
tantly 5-hydroxymethylfurfurals (5-HMF). The formation of 5-HMF
also prone to the accumulation of humic matter, levulinic acid, formic
acid and other by-products [190]. The mechanism of the formation of
5-HMF from hexose, particularly glucose (hydrolysis product of starch)
H. Zabed et al. Renewable and Sustainable Energy Reviews 71 (2017) 475–501

Table 9
Recent investigations for bioethanol production from starchy biomass.

Biomass Objectives Major findings Reference

Corn • Acorncomparative characterization of four normal and four high sugary • Corn genotypes varied in their agronomic traits, irrespective of the
genotypes grown for dry-grind ethanol production with regard groups of genotypes, and high sugary genotypes were not
[150]

to agronomic performance and kernel biochemical composition particularly unusual compared with normal genotypes
• High sugary genotypes were marked for higher sugars and lower
starch in the kernels than those of normal genotypes
• Evaluation of enzyme consumption and ethanol yield for normal and
high sugary corn genotypes during dry-grind ethanol production
• High sugary genotypes required significantly lower quantity of
enzymes and produced higher amounts of ethanol than normal
[35,151]

genotypes
• Six corn varieties were studied for simultaneous production of • The yield of two biofuels varied in response to the processing route,
bioethanol and biodiesel using four sequential steps, such as where maximum bioethanol yield was obtained from M-F-T route,
[152]

milling (M), germ separation (S), fermentation (F) and in situ and S-T-F route was marked for the highest biodiesel yield
transesterification (T)
• Study of 222 corn samples to predict dry-grind ethanol yield using
near infrared spectroscopy
• The selected wavelengths for predicting ethanol yield were
concentrated on 400–550 nm of the visible region and 2300–
[153]

2360 nm of the NIR region

• corn
Investigation of the effect of stillage recycling during fermentation of • Ethanol yield was not affected by the stillage recycling [154]
mash • Average theoretical ethanol yield was 83.38%
Sweet
sorghum
• Two-step conventional hydrolysis and fermentation of three sweet • Starch hydrolysis efficiency was 87–88% when a pre-gelatinization
sorghum cultivars containing different amounts of tannin ( < 0.02%, done prior to hydrolysis
[40]

1.1% and 2%) in the grains • Addition of a protease before adding α-amylase increased
• Effect of a pre-gelatinization on hydrolysis; effect of protease addition hydrolysis of starch
on hydrolysis; effect of tannin removal fermentation • Removal of tannin improved the fermentation efficiency
• Ethanol yield was 390 L/ton of dry grain
• Investigation of whole and decorticated sorghum grains using both • Decortication increased the productivity of fermentable sugars
conventional and granular starch hydrolysis process for evaluating during granular starch hydrolysis
[155]

enzyme consumption and quality of DDGS
• Aduedecrease in ethanol yield was observed when grains decorticated
to nutrient lost
• Enzymes consumption decreased by 11.7% when decorticated
grains were used during granular starch hydrolysis
• DDGS showed a decrease in neutral and acid detergent fiber
content when decorticated grains were used

Babassu cake • Evaluation of babassu cake (an agro-industrial waste) for generating • The enzyme system generated a hydrolysate that was rich in carbon
a multi-enzyme system and hydrolyzing the starch molecules and nitrogen nutrients
[106]

• Ethanol production by simultaneous saccharification and • Ethanol fermentation efficiency was as much as 83% with an
fermentation of the babassu flour ethanol concentration up to 59 g/L
Iles-iles tuber • Evaluation of Iles-iles tuber for ethanol production, which was a non- • Iles-iles tuber produced a sugar concentration of 140 g/L during
food starchy feedstock hydrolysis
[46]

• Ethanol concentration ranged from 50 to 80 g/L during

fermentation of the hydrolysate of Iles-iles tuber
Sweet potatoes • Bioethanol production from sweet potatoes through co- • Co-immobilization of yeast with either A. oryzae or M. purpureus
immobilization of saccharolytic molds (Aspergillus oryzae and generated 3.05–3.17% of ethanol
[156]

Monascus purpureus) and the traditional yeast (S. cerevisiae) • Co-immobilization of yeast with a mixed cultures of A. oryzae and
M. purpureus at a ratio of 2:1, generated 3.84% ethanol
• Co-immobilization of yeast with A. oryzae and M. purpureus at 1:2
ratio produced 4.08% ethanol
• onEvaluation the effects of post-harvest practices and heating methods • Ripening of the crop had significant effect on ethanol yield, and the
the overall conversion efficiency and ethanol yield of sweet highest yield was recorded when the materials were fermented after
[157]

potatoes 25 days of the harvest

• The conventional heating method was slightly superior to the
microwave method, showing a 9% higher efficiency by the former

Wheat meal • Bioethanol production from wheat

saccharification and fermentation
meal through liquefaction, • Significant amounts of maltose was generated during liquefaction
that reduced over time for simultaneous sachharification and
[158]

fermentation
• Theoretical ethanol yield varied between 87% and 89%
• Comparative bioethanol production from pure starch (PS), sterile
flour (SF) and unsterile flour (UF)
• Maximum theoretical conversion efficiency was found to be 96% for
PS, 69% for SF and 35% for UF
[95]

Rice wine cake • Evaluation

hydrolysis
of the rice wine cake without cooking of the slurry prior to • An ethanol fermentation efficiency was found to be 94.0% [159]

Carrot must • Optimization

fermentation
of the enzymatic hydrolysis of carrot must for ethanol • Optimum conditions for enzymatic hydrolysis were found as the
temperature 70 °C, pH-5.5, and time 2.5 h
[160]

• Ethanol production from the must was 77.5 L/ton under the
optimum conditions
Horse gram
flour
• Evaluation of the horse gram flour as a supplemented material for the • Ethanol yield increased considerably (up to 50%) after
high sugar containing broth during ethanol fermentation supplementation of broths with 4–6% horse gram flour
[161]

is shown in Fig. 8. The by-products produced during acid hydrolysis of rate of conversion during hydrolysis by the temperature and pressure
starch are toxic to microbial cells and inhibit the growth of yeast extrusion method, results in an inefficient conversion of starch and
affecting the overall yield of ethanol fermentation [191]. Likewise, the production of low amounts of fermentable sugars [192]. On the other
catalytic inhibitory effect of high pressure on α-amylase and the slow hand, enzymatic hydrolysis is of biological origin, eco-friendly, simple,

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H. Zabed et al.
Fig. 5. Schematic diagram of ethanol production from lignocellulose (adapted from
[129]).
specific, and an efficient method to convert starch into fermentable
sugars [42].
There are two basic steps in the conventional enzymatic hydrolysis
of starch, such as liquefaction and saccharification [42,151].
Liquefaction is normally done at high temperatures (85–105 °C in
laboratory or up to 165 °C in commercial plant) using thermostable α-
amylase [193], while saccharification is carried out at a relatively lower
temperature (50–60 °C or even at fermenting temperature, such as
30 °C) using glucoamulase [194]. During liquefaction, starch is first
gelatinized at a high temperature. The completeness in starch gelati-
nization is important as it will influence the conversion of starch into
fermentable sugars in the subsequent step. Almost all the amylose is
solubilized and leached out from the starch granules during gelatiniza-
tion and results in an increased viscosity of the slurry as it swells up at
this time. The α-amylase used is an endoenzyme and randomly cleaves
α-1→4 glycosidic linkages in starch molecules. Gelatinization and
liquefaction of starch polymer usually convert the starch polymer into
shorter chains, namely dextrins, maltose and maltotriose. The enzyme
glucoamylase then work on the liquefied starch during saccharification
and results in the conversion of dextrin into glucose. The heat-stable α-
amylase is obtained from thermophilic bacteria, such as Bacillus
licheniformis or from recombinant strains of Escherichia coli and
other Bacillus strains, whereas, glucoamylase can be obtained from
Aspergillus niger or Rhizopus species [195,196].
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9.2. Hydrolysis of holocellulose
The cellulose and hemicellulose of the lignocellulosic biomass are
required to convert into fermentable sugars before fermentation
through a process called hydrolysis [197]. In most cases, pretreatment
of the biomass produces two fractions, a water insoluble solid portion
containing mainly cellulose and lignin, and a liquid fraction containing
hemicellulose and its sugars. Based on the pretreatment methods and
conditions applied during pretreatment, hemicellulose of the biomass
may be either completely hydrolyzed to its monomeric sugars or
converted into oligosaccharides if it undergoes incomplete depolymer-
ization that requires a further hydrolysis [128,198]. On the other hand,
almost all of the cellulose remains unconverted under the pretreatment
conditions and need to go through hydrolysis process for converting
into glucose. Like the hydrolysis of starch, conversion of cellulose and
hemicellulose can be done using both acid and enzymes.
Acid hydrolysis of the pretreated biomass can be done via two
approaches, dilute acid treatment and concentrated acids treatment.
The former method is done at high temperature and pressure requiring
a short reaction time that ranges between seconds and minutes, while
concentrated acid treatment can be done at low temperature [33,199].
Mostly sulfuric acid (H2SO4) is used in either approaches of acid
hydrolysis, although other inorganic acids as well as some weak organic
acids such as hydrochloric acid (HCl), nitric acid (HNO3), trifluoracetic
acids (TFA), phosphoric acid (H3PO4) have also been reported to use
for this purpose [128]. The major drawbacks of the acid hydrolysis are
the requirements of high temperatures, requirements for either recov-
ery or neutralization of the acids and production of the inhibitors
through degradation of sugars.
Dilute acid is usually applied for the purpose of hemicellulose
hydrolysis and pretreatment of cellulose. However, both carbohydrate
polymers can be hydrolyzed with dilute acid that require a two-stage
hydrolysis process. The first stage is carried out at low temperature to
maximize hemicellulose conversion as hemicellulose is depolymerized
at lower temperature than cellulose. On the other hand, second stage is
conducted to convert cellulose into glucose at high temperature that
ranged between 230 °C and 240 °C [200,201]. The concentration of
acid normally used for dilute acid hydrolysis ranged from 0.5% to 1.5%
[33,128].
Concentrated acid hydrolysis can be applied for hydrolyzing both
hemicellulose and cellulose. The concentration of acid used for this
purpose ranged between 41% and 100% [202]. Reaction time required
for concentrated acid hydrolysis is generally much longer than the time
required for dilute acid hydrolysis. However, concentrated acid offers a
complete and rapid conversion of cellulose into glucose, and hemi-
celluloses into five carbon sugars [203]. Concentrated acid hydrolysis
has been shown effective hydrolysis method due to the moderate
process temperatures and not requiring enzymes [204]. However,
corrosion of the equipment with high concentration of acid is a major
drawback for this method.
Enzymes can hydrolyze both cellulose and hemicellulose specifically
to produce soluble sugars at low temperatures ranging from 45 °C to
50 °C and does not have any corrosion problem for the equipment
[205]. Compared to acid hydrolysis, sugar degradation and production
of inhibitors are much lower in enzymatic hydrolysis. Enzymatic
hydrolysis of cellulose and hemicellulose can be done by using cellulase
and hemicellulase (xylanase) systems, respectively.
Cellulase is a mixture of different cellulytic enzymes that act
synergistically on the cellulose molecules [206]. A typical cellulase
system includes endoglucanase, exoglucanase and β–glucosidase.
Endoglucanse exert its effect on cellulose through the random break-
down of β–1, 4 glycosidic linkages in the D–glucan chains from the
amorphous regions of a cellulose molecule that results in the produc-
tion of free chain ends. On the other hand, exoglucanase works on the
cellobiose units in D–glucan chains from the non–reducing ends. The
H. Zabed et al. Renewable and Sustainable Energy Reviews 71 (2017) 475–501

Table 10
Some recent investigations on lignocellulosic biomass for bioethanol production.

Feedstock Objectives Major findings Reference

Corn stover • Biological pretreatment of corn stover using Ceriporiopsis • Lignin degradation up to 31.59% [162]
subvermispora • Low cellulose loss (less than 6%)
• Incubation period 18 d • Maximum ethanol yield 57.80%
Corn cob • Co-generation of microbial lipid and bioethanol using corn cob • Ethanol yield 131.3 g/kg and lipid yield 11.5 g/kg under the [163]
• Acid hydrolysate of corn cob was used as substrate for microbial optimum conditions
lipid production, and remaining solid residue was enzymatically
hydrolyzed
• Conversion efficiency of fermentable sugars into valuable
products was 71.6%
Wheat straw • Bioconversion of wheat straw on a pilot scale • Maximum ethanol concentration 36 g/L [164]
• Major steps done by dilute acid pretreatment, bio-abatement of • Maximum ethanol productivity 0.43 g/L/h
fermentation inhibitors and simultaneous saccharification and • Maximum ethanol yield 0.29 g/g
fermentation using Escherichia coli FBR5 • Maximum theoretical ethanol yield 86%
• Effective fermentation time 83 h
Rice straw • Application of popping pretreatment technique on rice straw
for enhancing cellulose conversion efficiency
• Optimal
xylanase
enzyme load was 23 FPU/g for cellulase and 62 U/g for [165]

• Evaluation
enzyme dose
of the effect of popping pretreatment on the optimal • Sugar production under the optimum conditions was 0.567 g/g of
straw after 48 h
• Ethanol yield was 0.172 g/g of straw (80.9% of the maximum
theoretical) after 24
Sugarcane bagasse • Enzymatic hydrolysis of phosphoric acid impregnated and
steam exploded sugarcane bagasse under high solid load (18–
• Maximum
conditions
sugar concentration was 76.8 g/L under the optimum [166]

22%) and low enzyme load • Total glucan conversion was 69.2%
• sugarcane
Simultaneous saccharification and fermentation of delignified • Maximum ethanol concentration 25 g/L [167]
bagasse • Maximum ethanol productivity 0.7 g/L/h
• Cellulose conversion efficiency 72%
Sweet sorghum • Pretreatment and hydrolysis of sweet sorghum bagasse in a • Sugar yield 820 g/kg of biomass [168]
bagasse single step using microwave irradiation • Ethanol yield 480 g/kg of sugar after 24 h
Barley straw • Barley straw was treated with a novel method, such as • Maximum ethanol concentration 46 g/L [169]
treatment with sodium hydroxide in a twin-screw extruder for
continuous pretreatment
• Maximum ethanol yield 77.4%
• High biomass loading, up to 20%
Pine (soft wood) • Some important factors for the extrusion pretreatment method • The optimum extrusion conditions include a screw speed
were studied 150 rpm, barrel temperature 180 °C and moisture content 25%
[170]

• Maximum cellulose, hemicellulose and total sugar recoveries

65.8, 65.6% and 66.1%, respectively
Yellow poplar (hard
wood)
• Optimization of organosolv method for biomass pretreatment • Maximum ethanol concentration 42.8 g/L) under the optimum
using sulfuric acid pretreatment conditions (temperature 152 °C, acid concentration
[171]

1.6% and reaction time 16 min)

Water hyacinth • Optimization of pretreatment (Alkaline–oxidative) biomass and • The effective pretreatment conditions included 7% NaOH and 2%
saccharification of pretreated materials for bioethanol (w/v) H O at 100 °C
[172]

•
2 2
production Maximum ethanol yield 0.35 g/g of biomass
Switchgrass • Stem explosion of switchgrass for combined ethanol and • Removal of lignin was minimum (12%) when steam was used
methane production alone
[173]

• Comparison of ethanol yield under steam alone, steam with • Maximum removal (35%) of lignin was obtained when steam with
lime (calcium hydroxide) and H SO pretreatment lime used
• CH
2 4
Low lime increased the final ethanol yield, while H SO enhanced
2 4
yield
• Investigation of hemicellulose fraction of cocksfoot grass • The high severity conditions of pretreatment, such as at 180–
4
Cocksfoot grass [174]
(Dactylis obtained from wet explosion pretreatment 190 °C inhibited fermentation completely due to high
glomerata) concentration of inhibitors
• Maximum theoretical ethanol yield was 92% under the
pretreatment conditions 160 °C, 15 min, 87 psi oxygen
Municipal solid waste
(MSW)
• Pretreatment of some biodegradable MSW with fifteen
methods, followed by hydrolysis of the pretreated biomass
• Maximum glucose yield was 72.80% under a pre-hydrolysis
treatment conducted with 1% H SO , followed by steam
[175]
2 4
treatment at 121 °C, and enzymatic hydrolysis with Trichoderma
viride cellulase at 60 FPU/g of substrate

outcomes of both endoglucanse and exoglucanse action is production of
a disaccharide, namely cellubiose, which is finally converted into
glucose with the action of β–glucosidase [207]. An efficient cellulase
system should be capable of degrading cellulose in its crystalline stage,
along with showing tolerance to acidic pH (4–5) and stress conditions
[208].
Unlike cellulose, hemicellulose (xylan) is chemically quite complex
and its degradation requires multiple enzyme systems, which is
referred to as the xylanase (hemicellulase) system. Typically a xylanase
systm contains endo–xylanase, exo–xylanase, β–xylosidase, α–arabi-
nofuranosidase, α–glucoronisidase, acetyl xylan esterase, and ferulic
acid esterase [209]. Endo and exo–xylanase act on the main chains of
xylans and hydrolyze them into smaller chains. A β–xylosidase attacks
xylo-oligosaccharides and produce xylose. The α–arabinofuranosidase
and α–glucuronidase act on the xylan backbone and remove arabinose
and 4–o–methyl glucuronic acid, respectively [210]. The acetyl
esterases attack the acetyl substitutions on the xylose moieties, while
feruloyl esterases hydrolyze the ester bonds located between arabinose
substitutions and ferulic acid. Feruloyl esterases also make it easy to
release hemicellulose from lignin [119].
10. Fermentation of the sugars
Fermentation is a metabolic process of microorganisms that con-
verts soluble sugars into alcohol. Some bacteria and yeasts can
metabolize monosaccharaides (e.g., glucose and fructose) and disac-
charides (e.g., maltose and sucrose) in the absence of oxygen, which
results in the production ethanol and carbon dioxide [211]. The main

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H. Zabed et al. Renewable and Sustainable Energy Reviews 71 (2017) 475–501

Table 11
Advantages and disadvantages of different pretreatment methods (Adapted from [129,178].

Category Pretreatment method Advantages Disadvantages

Physical Mechanical grinding • Break the structure of lignocellulose • High energy input
• Reduce cellulose crystallinity • Not economically feasible
• Increase surface area
Irradiation • Can be combined with other treatment • Economically non-viable
• Increase surface area • Slow rate of reaction
• Improve glucose yield • Energy intensive
• Not eco-friendly
Chemical Dilute acid • Significant improvement of cellulose hydrolysis • Little lignin removal
• High reaction rate and high yields • Requirement of washing the pretreated biomass and
• Less corrosion problem and inhibitor formation neutralization before enzymatic hydrolysis
compared to concentrated acid pretreatment • Requirements of disposal of neutralization salts
• Most widely used method in industrial application
Concentrated acid • Complete removal of cellulose crystalline structure • Formation of inhibitors.
• Achievement of amorphous cellulose • Corrosion of equipment
• High reaction rate • Need for acid recovery
Alkali • Operation at low temperature • Conversion of alkali to irrecoverable salts or incorporation
• Alteration in lignin structure as salts into the biomass
• Lower degradation of sugars compared to acid
pretreatment
• Necessity of pH adjustment
Ozonolysis • cellulose
Selective lignin degradation with minimal effects on
and hemicellulose
• Highly
of ozone
reactive, flammable, corrosive and toxic properties

• Low inhibitor formation • High generation costs due to large energy demand
• Operation at ambient temperature and pressure • Exothermic
systems
characteristics of process may require cooling

Ionic liquids • No toxic or explosive gases are formed • Expensive

• Minimum degradation of desired products • Requirement of washing before reuse
• Operation at low temperature • Lack of mature commercial IL recovery methods
Organosol • Recovery of relatively pure lignin as a by–product • Requirement for removal of solvent from the system
• Minimum cellulose loss (less than 2%) • High costs of chemicals
• Low sugar degradation • Formation of inhibitors
Physicochemical Steam explosion • Partial hydrolysis of hemicelluloses • Less effective for soft wood
• Increases accessible surface area • Partial degradation of pentoses and lignin
• Decrease in the degree of polymerization • Loss of yield and formation of inhibitors
• Cost effective
• No significant environmental constraints
• Industrially developed
Acid catalyzed steam • Removal of hemicellulose • Formation of inhibitors
explosion • Increased surface area • Produce reactive loss and toxicity
• Increased enzyme accessibility
• Low environmental impact
Liquid hot water • Does not require any catalysts or chemicals • High water demand
• Increased accessible surface area • High energy requirement
• No or little inhibitor formation • Hemicellulose degradation
Ammonia fiber explosion • Increased surface area • Environmental concerns
(AFEX) • Reduction in cellulose crystallinity • Large amount of ammonia increase cost
• Removal of lignin • No utilization beyond lab scale
• No inhibitors formation
Ammonia recycling • Removal of lignin • High energy requirement
percolation (ARP) • Reduction in cellulose crystallinity • Environmental concern
• Alteration of lignin structure
• Low inhibitor formation
Soaking aqueous ammonia • Formation of low inhibitors • Environmental concerns
(SAA) • Low energy requirements
Wet oxidation • Increased digestibility of cellulose • High costs for oxygen and catalysts
• Formation of low inhibitors • Requirement of high pressure and temperature
• Removal of lignin
CO2 explosion • Effective removal of lignin • Requirement of costly equipment
• Low sugar degradation
• Low temperature
Biological Brown rot, white rot and soft • Reduction in the degree of polymerization of cellulose • Very slow rate of degradation and delignification
rot fungi and hemicellulose • Loss of carbohydrates as consumed by microbes
• No chemical required • Long residence times (10–14 days)
• Mild environmental conditions
• Low capital costs
• Low energy demand
• Low inhibitor formation
feedstocks in 1st and 2nd generation ethanol fermentation are sugar of glucose, fructose and maltose. The hydrolysates of starchy feedstocks
juice (sugary feedstocks) and hydrolysates of starch and lignocellulosic contain mainly glucose or a disaccharide (maltose), while the hydro-
materials. The sugar juice mostly contains sucrose with small amounts lysates of lignocellulosc biomass has both pentose and hexose sugars.

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H. Zabed et al.
Fig. 6. Biochemical components of lignocellulosic biomass and their potential degradation products (Adapted from [186,187]). Dashed line denotes the secondary degradation
products.
The traditional yeast used for ethanol fermentation can ferment
glucose and fructose, but not the pentose sugars. Disaccharide is first
hydrolyzed into its monosaccharides by the action of yeast enzymes. In
a typical yeast fermentation, glucose (a hexose sugar) is converted into
ethanol according to the reaction shown in Eq. (1). From this reaction,
the theoretical ethanol yield can be calculated. As per the Eq. (1),
100 kg of glucose will produce 51.1 kg of ethanol and 48.8 kg of CO2
[212].
C6 H12 O6 + H2 O + Yeast → 2CO2 + 2C2 H5OH + H2 O + Heat
Glucose Water Carbon Ethanol
dioxide
(100 kg) (48.8 kg) (51.1 kg) (1700 BTU)
(1)
During ethanol fermentation, roughly 95% soluble sugars are
converted into ethanol and CO2, 1% is converted into cellular matter
of the yeast cells, and 4% is converted into other soluble byproducts
such as glycerol [213]. The costs for yeast in ethanol production
account for nearly 10% of the total production costs [214].
Compared to the fermentation of starch hydrolysate that contains
only glucose, the hydrolysate of lignocellulosic biomass contains a
mixer of pentose and hexose sugars. Hexose sugars, such as glucose,
galactose and mannose are readily fermented by many naturally
occurring microorganisms, traditionally Saccharomyces cerevisiae
Fig. 7. Detoxification methods of lignocellulosic hydrolysates (Adapted from [185]).
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Renewable and Sustainable Energy Reviews 71 (2017) 475–501
and Zymomonas mobilis. However, natural microorganisms cannot
ferment pentose sugars, e.g., xylose and arabinose. On the contrary,
several strains of yeast, including Pachysolen tannophilus, Pichia
stipitis, Kluyveromyces marxianus, and Candida shehatae ferment
xylose [215], even though oxygen uptake rate of these strains may act
as the limiting factor for the conversion of xylose into ethanol
[216,217]. As attempts to overcome the oxygen limiting problem as
well as to ferment both xylose and glucose, a co-culture system has
been reported that used a xylose and a glucose fermenting yeasts,
namely P. stipitis and S. cerevisiae, respectively [218].
Fermentation can be performed by three modes, such as batch, fed–
batch and continuous mode [219]. Selection of an appropriate mode
for fermentation depends on the kinetic properties of the microorgan-
ism and type of substrate, in addition to the consideration of process
economics [220].
Batch mode of fermentation is carried out in a closed culture
system. The fermentation medium is supplemented with necessary
ingredients at the initial stage and yeast or other bacterial inoculum
added to this medium before starting fermentation. At the initial
stages, growth of yeast cells shows a lag phase that follows an
exponential phase afterwards. However, the cells compete for limited
amounts of nutrients in the medium and finally enter into a stationary
phase as nutrients are exhausted [33]. It is the simplest mode of
fermentation and does not require anything to add in the medium,
H. Zabed et al. Renewable and Sustainable Energy Reviews 71 (2017) 475–501

Fig. 8. Mechanism of the formation of 5-HMF from the hexose (glucose) obtained from starch hydrolysis (Adapted from [190].

except acid or base solutions for pH adjustment during fermentation.
In this situation, yeast or bacterial cells work under high substrate
concentration at the initial stage, while they need to show the activities
under a high ethanol concentration at the final stages [221].
When an amount of fermented media is removed from the fed-
batch system at an interval and the remaining part is used as inoculum
for the next culture, it is then referred to as the repeated fed-batch
fermentation. The fed-batch system combines both batch and contin-
uous modes of fermentation in technological perspectives and is widely
used for ethanol production on a commercial scale [222]. It is a batch
culture system that fed continuously or sequentially with the substrate
or other ingredients without removing the fermented broth. Microbial
cells grow at a low substrate concentration in this system throughout
the fermentation period and produce enhanced amounts of ethanol
during the course of fermentation. Fed-batch system offers some
advantages over batch mode, such as higher ethanol yield and
productivity, higher viable cells, longer life time of the cells, lower
concentration of residual sugars, shorter fermentation time, higher
dissolved oxygen in the mash, and lower toxicity on the cells [223].
Furthermore, it maintains the process conditions (temperature, pH,
and dissolved oxygen level) at specific levels through a feedback control
[33]. Nevertheless, the fermentation in a fed–batch system is limited by
the feed rate, where feed rate is limited by the concentration of the cell
mass [185]. The major advantages of this system are the non-
requirement of the fresh inoculum for each fed-batch and lower
contamination of the mash [223].
Continuous fermentation is conducted through a sequential input of
the required ingredients and removal of the products from the
fermentation vessel continuously. In other words, the feed containing
substrate, culture medium and nutrients, is pumped continuously into
an agitated vessel [33]. It can be performed in two kinds of bioreactors,
such as stirred tank reactors (single or series) or plug flow reactors.
Continuous fermentation often gives a higher productivity than batch
or fed-batch fermentations. It gives high productivity of ethanol and
requires short down time for vessel cleaning and filling [224]. However,
it is a more complex mode of fermentation and suitable for fermenta-
tion on large scale rather than laboratory experiments.
yeast, namely Saccharomyces cerevisiae [226]. Usage of this yeast in
the fermentation is one of the oldest practices in biotechnology, where
it has been used for producing drinking alcohol. Today, fermentation of
sugars using S. cerevisiae is widely used for generating fuel ethanol
from a variety of biomass. Several unique properties of this yeast have
made it most attractive for ethanol fermentation. Firstly, it has shown
greater efficiency in the conversion of sugars into alcohol and tolerance
to high concentration of ethanol [227]. Secondly, it has the capability
for producing flocs during its growth in the fermentation media,
making it easier to settle down or suspend when required [228].
Thirdly, it is a generally recognized as safe (GRAS) microorganism
[229]. Moreover, fermentation of some crop juices containing sucrose
is often carried out using yeast for its ability to hydrolyze sucrose into
glucose and fructose through secretion of invertase.
The metabolic pathway used by S. cerevisiae during ethanol
fermentation from sugars, particularly glucose, is the glycolysis.
During glycolysis in the yeast cells cytoplasm, each mole of glucose is
catabolized by different enzyme systems and finally generate two moles
of pyruvate (Fig. 9). In the absence of oxygen, each mole pyruvate
undergoes a reduction reaction that results in the generation of one
mole ethanol and one mole carbon dioxide, with a grand total of two
moles ethanol and two moles carbon dioxide from two moles of
pyruvate. A net of two ATPs are produced during glycolysis that are

11. Role of microorganisms

A variety of microorganisms are used in the conversion of biomass

into ethanol, which have effective roles on the overall conversion
process, including pretreatment, hydrolysis, detoxification and fermen-
tation. However, the performance of these bio-agents varies signifi-
cantly among the groups, species and strains of the microorganisms
with regard to the desired yield. A typical ethanologenic microorganism
should have some important characteristics, such as (1) good growth in
Fig. 9. Glucose metabolism by S. cerevisiae during fermentation towards ethanol
simple and inexpensive media, (2) high ethanol yield ( > 90.0% of
production [230]. Abbreviations: HK: hexokinase, PGI: phosphoglucoisomerase, PFK:
theoretical), (3) tolerance to high ethanol concentration ( > 40.0 g/L),
phosphofructokinase, FBPA: fructose bisphosphate aldolase, TPI: triose phosphate
(4) good ethanol productivity ( > 1.0 g/L/h), and (5) ability to retard isomerase, GAPDH: glyceraldehydes-3-phosphate dehydrogenase, PGK: phosphoglyce-
contaminants from growth condition [225]. rate kinase, PGM: phosphoglyceromutase, ENO: enolase, PYK: pyruvate kinase, PDC:
The most common microorganism used in ethanol fermentation is a pyruvate decarboxylase, ADH: alcohol dehydrogenase.

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H. Zabed et al.
Fig. 10. Carbohydrate metabolism by Z. mobilis during fermentation towards ethanol production [230,235]. Abbreviations: LEVU: levansucrase, INVB: invertase, GFOR: glucose–
fructose oxidoreductase, FK: fructokinase, GK: glucokinase, GPDH: glucose-6-phosphate dehydrogenase, PGL: phosphogluconolactonase, EDD: 6-phosphogluconate dehydratase,
KDPG: 2-keto-3-deoxy-6-phosphogluconate, EDA: 2-keto-3-deoxy-gluconate aldolase, GNTK: gluconate kinase. See Fig. 9 for PGI, GAPDH, PGK, PGM, ENO, PYK, PDC and ADH.
utilized by the yeast cells to biosynthesize the essential macromolecules
for growth of cells.
Another well-known microorganism used for ethanol fermentation
is a Gram-negative bacterium, Zymomonas mobilis, which has been
extensively studied over last several decades for ethanol fermentation
from starch and lignocellulosic hydrolysates, and raw sugars and
sucrose containing juices [231]. Compared S. cerevisiae, Z. mobilis
has shown higher tolerance to ethanol, higher glucose uptake, and
enhanced ethanol yield and productivity [230]. Nevertheless, because
of its narrow substrate range, Z. mobilis cannot immediately replace S.
cerevisiae in fuel ethanol production. Although nearly 2% of the carbon
source is usually converted into biomass [232], the biomass yield for Z.
mobilis in the fermentation media is generally low. More importantly,
cell growth and fermentation rate are not correlated during fermenta-
tion with this bacterial species [233], even though other organisms may
also show the same phenomenon depending on the substrate and
fermentation conditions. This is due to the fact that Z. mobilis
metabolize glucose using Entner–Doudoroff pathway that yield only
one molecule of ATP from each molecule of glucose (Fig. 10). As a
result, the molar growth yield is the minimum for Z. mobilis compared
to those of other fermenting bacteria [234].
Although many natural microorganisms are used in ethanol fer-
mentation on the laboratory scale, there are some limitations with
these bio-agents for exploiting on commercial scale. On the other hand,
the widely used yeast, S. cerevisiae and the promising bacterial species,
Z. mobilis can utilize only soluble sugars (glucose, fructose and
sucrose). Therefore, an additional step is required for converting
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polymeric carbohydrates (starch, cellulose and hemicellulose) into
fermentable sugars using a large quantity of enzymes that results in
an increase in the production costs and process time [236,237].
Another concern with natural microorganisms is their incapability to
ferment both hexose and pentose sugars, which are the two major
fermentable sugars obtained from cellulose and hemicellulose hydro-
lysis, respectively [238]. Furthermore, the efficiency of the natural
microorganisms for producing enzymes to hydrolyze polymeric carbo-
hydrates is also economically unattractive, in addition to low yield and
poor quality of the enzymes. To overcome these hurdles, novel
microorganisms have been developed [239] through alteration in the
genetic patterns of the natural microorganisms with the desired traits
through either recombinant DNA technology [128,240,241] and/or
evolutionary engineering techniques [242–244].
As attempts to convert starch into ethanol directly, recombinant
yeast strains have been developed successfully through the cell surface
displaying engineering that are capable of producing α-amylase or
glucoamylase or even co-expressing of both amylolytic enzymes,
resulting in the production of ethanol from native starch without
requiring separate liquefaction and saccharification [245–248].
Likewise, the genetic modification in natural bacteria and yeast cells
has been studied extensively to generate new strains that are capable of
co–fermenting both pentose and hexose sugars with high yields [199].
For example, Escherichia coli KO11 [249] and Klebsiella oxytoca P2
[250] recombinant strains have been developed that can ferment both
xylose and glucose.
H. Zabed et al. Renewable and Sustainable Energy Reviews 71 (2017) 475–501

Table 12
Some technological approaches investigated for bioethanol production from different biomass.

Major technology Feedstock Major focus Ethanol fermentation Reference

kinetics

VHG and SSF Potato tuber Utilization of high solid load (280g/L) in SSF P=117.72 g/L [251]
Q=1.96 g/L/h
Y=91% of theoretical
SSF Corn meal Ultrasound assisted liquefaction prior to SSF P= 9.67% w/w [252]
Q=3.02 g/L/h
Y=88.96% of theoretical
SSF Wheat straw Fermentation of steam-exploded wheat straw using a thermotolarant P= 36.2 g/L [253]
yeast strain in SSF
SSCF Wheat straw SSCF of steam pretreated wheat straw P= 43.7–53.0 g/L [254]
VHG and SSF Potato mash Utilization of high carbohydrate load (304 g/L) in SSF P= 131.05 g/L [255]
Q=2.13 g/L/h
Y= 89.7% of theoretical yield
VHG Sweet sorghum juice Fermentation of juice containing 280 g/L of total sugar P= 120.68 g/L [256]
Q= 2.01 g/L/h
Y= 0.51 g/g
GSHSF Sweet potato Cold SSF of the slurry containing 200 g/L initial solid using GSHE P= 21.6 g/L [257]
Immobilization and SSF Corn meal Immobilization of yeast cells (S. cerevisiae var. Ellipsoideus) on the P= 10.23% (w/w) [258]
calcium alginate and use of immobilized cells in SSF Q= 2.13 g/L/h
Y= 98.08% of the theoretical
yield
Immobilization Sugarcane juice and Immobilization of S. cerevisiae AS2.1190 cells on sugarcane pieces and P= 77.13–89.73 g/L [259]
molasses fermentation of juice and molasses with immobilized cells Q= 2.48–2.62 g/L/h
Y= 90.11–94.28% of the
theoretical yield
Immobilization Corn stover Fermentation of hemicellulose hydrolysate using immobilized cells of a P= 30.1–31.1 g/L [260]
recombinant yeast on calcium alginate Y= 0.39–0.41 g/g
Solid-state fermentation Sweet sorghum stem Advanced solid-state fermentation of the stem on a demonstration scale Y= 90.5% of theoretical [261]
CBP Wood chips Direct ethanol production from woods using a wood rot fungus Y= 0.04–0.26 g/g [262]
(Schizophyllum commune)

GSHE= granular starch hydrolyzing enzyme; GSHSF= granular starch hydrolysis and fermentation; P= ethanol concentration; Q= volumetric ethanol productivity; SSCF= Simultaneous
saccharification and co-fermentation; SSF= simultaneous saccharification and fermentation; VHG= very high gravity fermentation; Y= ethanol yield

12. Technological approaches
Recent advancements in the bioethanol production mainly focus on
the technological improvements to achieve enhanced ethanol yield per
unit of biomass, decreased production costs and process time, and
more importantly, minimized process steps, particularly during starch
and lignocellulosic ethanol production. Among these technological
approaches, the mention worthy achievements are the development
of a non-cooking hydrolysis technique for starch based feedstocks, cell
immobilization, development of recombinant microorganisms, con-
ducting fermentation under high solid load conditions, development of
solid state fermentation, and integration of different process steps.
Table 12 summarizes the investigations and achievements of some
technological approaches.
12.1. Non-cooking method for starch hydrolysis
High energy consumption during conventional ethanol production
hinder the production of cost-effective and eco-friendly ethanol [263].
It has been estimated that 30–40% of the total energy required for
ethanol production is consumed by cooking and liquefaction [237].
What's more, thermal processes employed during enzymatic hydrolysis
may produce unwanted by products through Maillard reactions [264].
Recently, a non-cooking method has been introduced to decrease
energy consumption during ethanol production that includes hydro-
lysis of granular starch at sub-gelatinization temperature using a
granular starch hydrolyzing enzyme (GSHE) [265,266]. It does not
require any gelatinization and liquefaction of starch prior to fermenta-
tion unlike in the conventional technique. Both hydrolysis and
fermentation can be completed in a single step using a granular starch
hydrolyzing enzyme (GSHE). Non-cooking method has been consid-
ered more promising for dry–grind ethanol production as it offers some
advantages over the conventional method.
Firstly, GSHE can catalyze and hydrolyze starch polymers at sub-
gelatinized temperature due to having both α–amylase and glucoamy-
lase activities. It does not require any activators, such as Ca2+, as is
frequently used in a conventional enzyme system [264,267]. Therefore,
GSHSF simplifies the overall ethanol production process and reduces
energy consumption by 10–20% [268]. Secondly, gelatinization of
starch at high temperatures in a conventional method results in the
increase in viscosity of the slurry by 20 fold, thereby causing difficulties
in mixing and pumping of the hydrolysates [268,269]. Furthermore the
increased viscosity may hinder the dispersion of starch and enzyme
molecules throughout the mixture, particularly under high dry solid
conditions and result in the incomplete conversion of starch.
Alternatively, viscosity problems can be overcome using non-cooking
method as it does not require any gelatinization of starch. Moreover,
non-cooking technique is suitable for using high solid load in the
slurries (12–38%) without encountering any mixing problems with
enzymes and substrates [270,271].
Compared to the conventional dry–grind ethanol process, non-
cooking dry-grind process produces a higher amount of ethanol,
because less sugar is lost during hydrolysis as often occurs in the
conventional process through the Maillard reaction. Additionally, this
method decreases osmotic stress on yeast cells during fermentation,
and produces more nutritious distiller's dried grains with solubles
(DDGS) [272]. It has been reported previously that higher amounts of
ethanol was produced with higher fermentation rates during dry–grind
ethanol production from raw corn using non-cooking technique
compared to the conventional method [273].
However, low hydrolysis rate and incomplete hydrolysis of starch at
sub-gelatinized temperature due to structural heterogeneity and crys-
tallinity of native starch have made the process further challenging
[274]. To overcome the shortcomings and increase the hydrolysis
efficiency as well as ethanol yield, enzyme manufacturers have recom-
mended to conduct a mild heat treatment (e.g. at 60 °C) prior to

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H. Zabed et al.
fermentation and to supplement the media with urea and protease
[275]. Urea has been reported to increase hydrolysis rate and yield
[274], and protease increase ethanol productivity and yields during
ethanol production from starch based feedstocks [276,277].
12.2. Cell immobilization
Microorganisms are frequently used as free cells for ethanol
fermentation, which proliferate in the media and carry out their
metabolic functions that result in the production of ethanol.
However, the specific growth rate of cells in a batch fermentation
system containing free cells can be affected by many factors related to
either product or substrate [278]. To overcome the substrate and
product inhibition on the cells as well as enhance the ethanol tolerance,
cell immobilization has been explored in the bioethanol research
during last several decades and still it has hold much interests in the
bioethanol research [279].
Immobilization of cells offers some technical and commercial
benefits over free cell system, such as prolonged cellular stability,
increased tolerance to high substrate concentration, increased ethanol
yield, greater volumetric productivity, reduced end product inhibition,
reduced risk of contamination due to high cell densities, decreased
energy demand and process expenses due to easier product recovery,
regeneration and reuse of cells for extended periods, cell recycling in
repeated batch fermentations, and protection of cells against toxic
substances [278,280].
The most common microorganism used in fermentation is S.
cerevisiae. Cells of this yeast can be immobilized via two basic
approaches, such as immobilization on supporting material and
immobilization by self-flocculation. Various supporting carriers have
been investigated thus far as a mean of immobilization on the
supporting materials that include apple pieces, k–carrageenan gel,
polyacrylamide, g–alumina, chrysotile, calcium alginate, sugarcane
pieces, banana leaf sheath, and orange peel [281]. The enriched culture
media of cells growing at the log phase can be harvested and entrapped
on the support to get beads with immobilized cells [282]. However,
entrapment strategy has a major drawback as it may difficult for the
slow growing cells in the system to come out effectively during
fermentation [230]. Self-flocculation of yeast cells is a non-sexual
and reversible cell aggregation, where yeast cells adhere to each other
and form a floc [283]. This technique is more promising than the
immobilization on the supporting materials as it is technically simple
and economically competitive, completely eliminates the contamina-
tion, offers cells to grow without being affected by environmental
factors, and gives the feasibility to control the desired concentration in
the reactors and recovery of yeast cells through purging the reactors
[230].
12.3. Very high gravity fermentation (VHG)
The assessment of bioethanol production with regard to energy
requirements has revealed that distillation is one of the most energy
consuming steps [284]. An increased concentration of ethanol in the
fermentation broth can considerably reduce energy consumption
during distillation process [285]. However, ethanol yield of a normal
gravity fermentation is usually low as the substrate concentration is
fixed. In the most cases, the range of initial solid in the fermentation
media is 20–25% for starch and 10–15% for ligocellulosic biomass.
Very high gravity (VHG) fermentation can use high concentration of
initial substrate and results in the production of increased amounts of
ethanol. This is a technique employed in fermentation of the processed
mash containing 270 g/L or more dissolved solids [256,286].
VHG has been extensively used during ethanol production from
cereal grains due to low viscous nature of cereal based feedstocks. As a
result, little investigations have been done using root or tuber crops as
they contain high amounts of pectin that increases viscosity of the
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slurry [251]. High viscosity in the fermentation media usually causes
resistance to solid-liquid separation and handling difficulties during
hydrolysis and fermentation resulting in incomplete hydrolysis of the
substrates as well as low fermentation efficiency [269,287]. To reduce
viscosity of the root and tuber feedstocks, excessive amounts of water
can be added. However, sugar concentration also diluted with the
addition of water that results in lower ethanol yield as reported only
3.9–8.8% ethanol in previous studies [288,289]. As attempts to reduce
the viscosity of a potato mash, Lim et al. [251] have treated it with a
combination of celulase, pectinase and viscozyme prior to subjecting to
enzymatic hydrolysis of its starch.
VHG technology exploits the enhanced and prolonged growth of
yeast in the presence of low level of oxygen [290]. The major
advantages of VHG are the increased fermentor throughput, reduced
water and energy consumptions, decreased labor and distillation costs,
reduced risk of bacterial contamination, and high ethanol yield
[220,251]. However, the application of VHG for ethanol fermentation
creates some problems. As high solid concentration is used in VHG, it
ultimately results in the presence of high initial sugars in the
fermentation broth. This high sugar concentration causes high osmotic
stress on the cells, which in turn, leads to increased cell lysis, reduced
cell viability and decreased fermentation rate [291]. The effects of high
osmotic stress can be reduced by supplementation of the fermentation
mash with some ingredients, which include, for example, ammonium
ions [256], urea [292], yeast extract [293], soya flour [294], and oilseed
meal [295]. These nitrogen or fat containing sources enhance the
growth rate and viability of the cells in the VHG media by releasing free
amino nitrogen or fatty acids, which results in the high ethanol yield
and productivity [291].
12.4. Solid state fermentation
Ethanol fermentation can be done via submerged or liquid state
fermentation and solid state fermentation, even though the former is
applied in almost all the fermentation practices. In the liquid state
fermentation of starch and lignocellulosic biomass, water is an
important liquid used to make slurry by mixing a pre-defined solid
with water, which results in the requirement of significant amounts of
water to complete the process. The necessity of the waste water
treatment after fermentation is another cost increasing factor in
submerged fermentation. Furthermore, liquid state fermentation of
sugar crops requires an extraction of juices and concentration it
through boiling or evaporation technique that increase the energy
consumption in the process [261].
On the other hand, solid state fermentation is the conversion of
biomass in its natural state, where growth and metabolism of cells
occurs on the moistened solids that results in the production of soluble
sugars and ethanol directly without requiring any free flowing water or
extraction of juice from sugar crops. The expected advantages of solid
state fermentation over the liquid state fermentation are [207,296].
• Lower requirement of water
• Lower requirement of energy for the physical treatment of biomass
• Smaller volume of the reactor
• Lower liquid waste production
• Lower capital investments
• Lower costs for product recovery and drying
• Lower sterilization energy costs
• Easier aeration due to a larger surface area
• Lower risks for bacterial contamination
`Nevertheless, solid state fermentation may have some bottlenecks
to be implemented it on large scale ethanol production [261]. Firstly,
the absence of water in the fermentation media during solid state
fermentation causes a decrease in heat transfer between the particles
and consequently, mixing of solid substrate particles is difficult.
H. Zabed et al.
Secondly, the control of mass and heat transfer is one of the major
challenges in the design and operation of reactors on large scale
fermentation. Thirdly, difficulties with the agitation in the substrate
particles due to high viscosity that leads to a heterogeneous mixing of
the feedstock particles, problem to control fermentation and difficulty
in feedstock management in the fermentation environment. Finally, the
growth of many fermenting microorganisms under low water condi-
tions is difficult. Consequently, it is still challenging to implement solid
state fermentation technique on commercial scale. As attempts to
overcome the above obstacles and successful introduction of this
promising technique on large scale, a special rotary drum reactor has
been designed in an advanced solid state fermentation that can
effectively control mass and heat transfer and produce higher amounts
of ethanol at short fermentation time [261,297,298].
12.5. Process integration
As stated earlier, the conversion of starch into ethanol requires two
major and key steps, namely hydrolysis and fermentation, while
lignocellulosic biomass necessitates thermochemical or biological pre-
treatment prior to subjecting to hydrolysis and fermentation. On the
other hand, ethanol can be produced from sugar based feedstocks
directly by fermentation without any pretreatment or hydrolysis step.
Therefore, one of the major improvements needed for starch and
lignocellulosic ethanol is the reduction of process steps to decrease
capital and operational costs as well as process time. Recent research
efforts have been made on the integration of pretreatment and
hydrolysis steps with fermentation in various ways that produce
enhance amounts of ethanol along with volumetric ethanol productiv-
ities [299,300].
Typically, enzymatic hydrolysis and fermentation of sugars are done
separately, and the process is known to separate hydrolysis and
fermentation (SHF). In a commercial SHF system with lignocellulosic
biomass, joint liquid flow from two hydrolysis reactors (hemicellulose
and cellulose) first enters the glucose (hexose) fermentation reactor.
Once fermentation completed, the broth of the reactor is distilled to
recover the bioethanol that leaves unconverted xylose and other
pentose behind. Subsequently, pentose fermentation is carried out in
a second reactor, followed by a further distillation of the fermented
broth [301]. After a slight modification, separate hydrolysis and co-
fermentation (SHCF) process comes out when both pentose and hexose
sugars are produced in separate hydrolysis of hemicellulose and
cellulose, and both sugars are fermented together [128]. The perfor-
mance of both cellulose hydrolysis and fermentation of hexose sugars
at a time is called simultaneous saccharification and fermentation
(SSF).
When SSF includes the co-fermentation of hexose and pentose
sugars, it is called simultaneous saccharification and co-fermentation
(SSCF). The consolidated bioprocessing (CBP) is another process
integration that involves not only the hydrolysis and fermentation of
all kinds of soluble sugars but also produce hydrolyzing enzymes,
resulting in the enzyme production, hydrolysis and fermentation in a
single step.
Among these process integrations, SSF has drawn particular
attention and widely used in both laboratory and commercial plants
over SHF to overcome some obstacles with the latter. In a typical SHF,
hydrolysis is done before fermentation. However, it has been reported
that enzymatic action can be inhibited by the accumulation of sugars in
the solutions as hydrolysis progresses, resulting in the incomplete
hydrolysis of carbohydrates [267]. In addition, a higher concentration
of sugars in the hydrolysate may cause osmotic stress on the yeast cells
[278], and as a consequence, ethanol productivity and final yields are
affected [302]. In comparison, SSF can avoid osmotic stress on the
yeast cells, end product inhibition on the enzyme and microbial
contamination during fermentation [255].
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13. Factors affecting conversion of biomass into ethanol
Various factors have been reported to affect the efficiency of
hydrolysis and fermentation of the biomass, which are mostly related
to substrate, enzyme, microorganism, and process parameters
[40,303,304]. The major factors include temperature, pH, incubation
time, initial solid load and microbial load, and accumulation of soluble
byproduct in the fermentation broth.
13.1. Temperature
Temperature is an important factor carefully regulated during
hydrolysis and fermentation as it has vital impact on the overall
conversion efficiency and product yields [305]. During conventional
hydrolysis of starch, the optimum temperature for liquefaction has
been reported to be 85–105 °C [193,194], while a separate sacchar-
ification is usually done at 50–60 °C [194,267]. On the other hand
starch hydrolysis in the non-cooking method is done at 30–35 °C
[269,274]. Cellulose hydrolysis is often done at a temperature ranged
between 45 °C and 50 °C [205].
The effectiveness of ethanol fermentation primarily relies on the
adequate growth of yeast cells in the broth that is often affected by the
temperature. It is generally believed that the ideal temperature for
fermentation ranges between 20 and 35 °C [306,307]. The optimum
temperatures for ethanol fermentation with free cells of S. cerevisiae
varied between 28 °C and 32 °C [308]. However, slihtly higher
temperature has been reported for immobilized yeast cells, probably
for the fact that immobilized beads transfer heat from particle surface
to inside the cells [309].
Ethanol yield can significantly be affected by the fermentation
temperatures, which to some extent, increases with the increase in
temperature. However, many earlier studies have shown that tempera-
tures above 37 °C are harmful for the growth of Z. mobilis and ethanol
fermentation [231,310,311]. Likewise, yeast cells are significantly
affected at temperatures above 40 °C [312,313]. Ethanol yield signifi-
cantly decreased at high temperature as a result of increased osmotic
stress, which further influence to produce high amounts of by-products
such as sorbitol and levan during fermentation [231]. High tempera-
ture is a stress factor for yeast cells, during which yeast produces heat-
shock proteins and inactivates its cellular ribosomes in response to the
stress condition [307]. Furthermore, a higher fermentation tempera-
ture might exert an indirect effect on the cells through the denaturation
of ribosomes and some essential enzymes, and creating problems with
the fluidity of the membranes [307,314]. On the other hand, much
lower temperatures during fermentation causes lower specific growth
rates of the cells and results in the lower tolerance to ethanol
[308,315].
13.2. pH
The pH of the solution is an important factor having significant
effects on the overall process, particularly during enzymatic hydrolysis
of starch and lignocellulosic biomass, and fermentation. The activity of
an enzyme is optimum up to a certain level of pH in the slurries that
varies depending on the type of enzymes and process employed. The
optimum pH ranged from 5 to 8 for α-amylase [316], 4–5 for
glucoamylase [156] and 4.5–5 for cellulase [60].
The pH is one of the key factors for ethanol fermentation that has a
direct effect on the yeast cells and cellular processes [257,317]. In
particular, the concentration of proton (H+) in the fermentation media
above or below a certain level can change the total charge on the
plasma membrane, thereby affecting the permeability of some essential
nutrients into the cells. Moreover, pH can also affect the fermentation
time. For example, a higher fermentation time has been reported for
producing maximum concentration of ethanol at a pH less than 4. On
the other hand, a higher pH above 5 may increase the production of
H. Zabed et al.
soluble by-products, such as glycerol and acetic acid that eventually
decrease final ethanol yield [318].
The optimum pH range in fermentation process may vary depend-
ing on the substrate used and strain of microorganism. The optimum
pH range for S. cerevisiae for ethanol fermentation has been reported
to be 4–5 [318], while a different pH range (5−6) reported for Z.
mobilis to get the best ethanol yield [319]. However, recently it has
been exceptionally reported that S. cerevisiae can produce ethanol
from date juice at low pH such as 3.8 [21]. As an example for showing
the effect of substrate on the optimum pH ranges, molasses can be
pointed out here that has a buffering capacity in the fermentation
media [231]. This buffering capacity of molasses further depends on its
chemical composition.
13.3. Incubation time
Incubation time is another factor that significantly affect enzymatic
hydrolysis and fermentation [265]. In general, extremely short or high
incubation time affects both sugar and ethanol yields. During enzy-
matic hydrolysis, enzymes usually diffuse into the slurry and get access
to the starch molecules, which then bring about the conversion of
carbohydrate polymers into fermentable sugars. The overall conversion
process requires an appropriate reaction time, where short incubation
period interrupts the hydrolysis process and results in the incomplete
conversion of carbohydrates. However, sugar yield shows a plateau
after a certain time of hydrolysis. This could happens due to an end
product inhibition to the enzymes after a certain time [267,320].
Short incubation time during fermentation causes inadequate
growth of yeast cells as well as inefficient conversion of sugars into
ethanol. On the other hand, high fermentation time causes toxic effect
on microbial growth, particularly in the batch mode due to the high
concentration of ethanol and production of soluble by-products [255].
It has been reported that the ideal fermentation period for ethanol
production is 48–72 h [321]. However, higher fermentation period
such as 96 h has also been reported, even though productivity of
ethanol significantly decreases as the fermentation time increases from
72 h to 96 h [142].
13.4. Initial substrate concentration
Initial substrate (sugar, starch, cellulose and hemicellulose) con-
centration significantly affects overall chemical and biochemical con-
version process. It has been reported that initial substrate concentra-
tion has direct effects on the rate of hydrolysis and fermentation as well
as the growth of yeast cells [267,304,322]. During enzymatic hydrolysis
of starch, cellulose and hemicellulose, excessive amounts of substrates
in the slurry may exert substrate inhibition of the enzymes that may
results in an incomplete conversion of carbohydrates and decrease in
sugar yield [267,323]. High substrate concentration creates a particular
problem during conventional cooking and liquefaction of starchy
materials that gelatinizes starch and increase the viscosity of the slurry,
making difficulties in mixing and pumping of the contents [268,269].
Furthermore, increased viscosity creates problem in the dispersion of
the starch and enzymes properly and results in the incomplete
conversion of starch [267,269]. Although this viscosity problem can
be overcome during granular starch hydrolysis, the optimum concen-
tration is still limited to 12–38% of initial solid [270,324].
High sugar concentration has direct effect on the fermentation rate
and growth of microbial cells during fermentation due to high osmotic
stress [325]. The actual relation between the initial substrate concen-
tration and the fermentation rate is rather more complex. In general,
fermentation rate increases with the increase in sugar concentration up
to a certain level [326]. But excessively high sugar concentration
exceeds the sugar uptake capacity of the cells that results in a steady
state in the fermentation. However, high initial sugar concentration is
often desired in a commercial plant for generating enhanced amounts
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of ethanol and decreasing the downstream processing costs. It takes
prolonged fermentation time and subsequently increases the recovery
costs, in addition to creating osmotic stress on the yeast cells.
Consequently, optimum initial substrate concentration is required for
an efficient conversion process.
13.5. Microbial load and accumulation of byproducts
Inoculum concentration of yeast or other fermenting microorgan-
isms does not have a significant influence on the final ethanol
concentration, but it significantly affects sugar consumption rate and
ethanol productivity [25]. However, it has been reported that ethanol
production increases with the increase in the initial cell numbers from
104 to 107 cells/ml, and there were no significant differences in ethanol
production when cell concentration ranged between 107 and 108 cells/
ml [281]. Nevertheless, increased cell concentration within a certain
range reduces fermentation time due to the rapid growth of cells and
immediate conversion of fed sugars into ethanol. In an investigation
with corn meal hydrolysate, it was found that fermentation time
dropped to 32 h from 48 h as the concentration of yeast inoculum
increased from 1.0% to 2.0% [267]. Likewise, a decrease in fermenta-
tion time has been reported from 72 h to 48 h when yeast concentra-
tion increased from 3.0% to 6.0% [327]. The increase in cell concen-
tration causes a decrease in the amounts of residual sugars left
unconverted during fermentation in a co-immobilization technique
[156].
One of the major challenges for controlled fermentation is the
bacterial contamination of broth, with a greater concern on lactic acid
bacteria [265]. Contaminant bacteria affect final ethanol yield and
productivity by competing with yeast cells for fermentable sugars,
nutrients (trace minerals, vitamins, and free amino nitrogen) and
producing inhibitory byproducts, such as lactic acid [328,329]. In order
to decrease bacterial contamination during fermentation, antibiotics
are often used in the fuel ethanol plants [330,331].
During ethanol fermentation, some soluble by-products such as
lactic acid, acetic acid and glycerol produce as a result of metabolic
activities of yeasts and contaminating bacteria [332]. Lactic and acetic
acids are usually produced due to the metabolism of carbohydrates by
the contaminating lactic acid bacteria. Production of lactic acid during
fermentation is unwanted as it affects yeast growth and ethanol yield
[154]. Glycerol is another byproduct produced by yeast cells or even by
the contaminating bacteria during fermentation [333]. Even though
production of glycerol is a part of sugar metabolism by yeasts, high
glycerol accumulation in the fermentation broth is undesirable.
Because production of glycerol is energy intensive process and affects
final ethanol yield [334].
14. Life cycle and economic analyses
Life cycle analysis (LCA) and economic analysis of bioethanol
generated from any renewable source are two important issues in the
overall sustainability of this promising alternative fuel. LCA is a
popular and widely used methodology for assessing the global envir-
onmental impact of any product, process or pathway with its partial or
whole life cycle generated from ‘‘cradle to grave”. LCA of a biofuel is
often limited to the balance of energy and/or greenhouse gas emissions
[335]. In recent years, LCA has been considered as a vital assessing tool
to qualify fuels under a number of regulations, including US Renewable
Fuel Standard, European Union Renewable Energy Directive and
California Low-Carbon Fuel Standard [336].
A good number of LCA has been analyzed till to date for first and
second generation bioethanol using various models, scopes, accuracy,
transparency and consistency levels [335–341]. However, most of
these analyses are not conclusive as the outcomes and interpretations
of different approaches vary considerably with regard to the model,
analysis tool, feedstocks and process conditions. It has recently been
H. Zabed et al.
reported that the results of LCA are greatly affected by the proportion
of bioethanol in the gasoline–bioethanol blend and by the source of
feedstocks, where the decrease in greenhouse gas emissions is less than
10% for E10 blend and higher than 40% for E85 and upper blends
[342]. In another recent study, the results of LCA have showed that a
vehicle driven one kilometer by E10 and E85 ethanol-gasoline blend
fuel could reduce the greenhouse gas emissions by 4.3% and 47% and
ozone layer depletion by 3% and 66%, respectively [338].
Economic analysis of ethanol production from renewable sources is
one of the major works that determine the overall costs for the process.
Many studies have been done till to date for economic analysis using
different feedstocks and models [95,343–345]. However, these ana-
lyses are not conclusive and vary depending on the feedstock,
technology and analysis technique. Total production costs of an ethanol
manufacturing process are the sum of several unique costs that include
raw material costs, operating costs, depreciation of capital, co-product,
and labor, supplies and overhead. The economic parameters for 1st and
2nd generation ethanol production facilities include, feedstock, plant
feed rate, plant type, location, annual fuel ethanol production, on-
stream days, and year for cost basis [345].
Among the total capital costs, raw material costs are significantly
high in sugar and starch based feedstocks compared with those of
lignocellulosic biomass. On the other hand, lignocellulosic biomass and
starchy feedstocks require enzymes during hydrolysis that increases the
overall capital costs for these biomass. In addition, the pretreatment
cost for lignocellulosic biomass further increases its capital costs. As a
result, the costs of ethanol generated from lignocellulosic biomass still
higher compared to the ethanol produced from sugars and starch. The
estimated costs for corn material used in a typical dry-grind process is
45–48%, while the operating costs ranges between 19% and 22% of the
total production costs [340]. The enzyme costs for a typical conversion
of woody biomass into fermentable sugars have been estimated to be
nearly 10–20% of the total capital costs [346].
15. Conclusion: future perspectives
Rapid depletion of fossil fuels and increased greenhouse gas
emissions have led to the development of alternative energy sources.
Bioethanol is one of the most promising and environmentally friendly
renewable sources for energy. Currently, sugarcane and corn together
produce almost all the bioethanol on the commercial scale. However,
these two feedstocks are not enough to meet the rising demand of the
bioethanol as well as replace huge amounts of conventional fuels
currently consumed. For this reason, recent research efforts have
focused the searching of potential ethanol sources, with a particular
attention on the lignocellulosic biomass. Improvement of the techno-
logical approaches and optimization of various factors have also been
given priority in the current bioethanol research. Nevertheless, there
are some challenges still needs to be properly addressed for developing
a sustainable bioethanol industry. Compared to the earlier stages,
higher amounts of ethanol is now being produced from sugarcane in
Brazil due to certain agricultural improvements that have resulted the
development of some new verities with increased biomass yields.
However, sugar yield from sugarcane has been plateaued to 140 kg/
ton of sugarcane during last 15 years, requiring a further effort for
increasing sugar accumulation in the sugarcane [347]. Breeding for
developing new crop varieties, planting techniques, harvesting time,
juice collection and preservation are some further challenges with the
sugar crops. With regard to the starchy crops, significant improvements
have been achieved in the overall conversion process; especially
ethanol production from corn is now a cost-effective and mature
technology. However, development and exploitation of the potential
corn hybrids and new non-food crops on commercial scale are still in
the immature stages. Requirement of costly and, in most cases, high
energy consuming pretreatment method for lignocellulosic biomass is a
major hurdle in the usage of these promising feedstocks on large scale
496
Renewable and Sustainable Energy Reviews 71 (2017) 475–501
ethanol production. Although some improvements have been made in
recent years through integrating the process steps, implementation of
these approaches on a pilot or commercial scale would be more
effective for developing a sustainable lignocellulosic ethanol industry.
Ethanol tolerance limit of yeast cells, osmotic stress on the cells by high
substrate concentration, acidity of the broth while fermentation
progresses and bacterial contamination during fermentation are some
issues need to be addressed for improved fermentation efficiency.
Screening and development of the stress tolerant strains would be a
promising option for overcoming stress conditions. Successful com-
mercial exploitation of recombinant microorganisms expressing the
pathways for pentose and hexose utilization, over expression of existent
pathways, increased ethanol generating pathways, deletion of by–
product producing pathways and expression of detoxifying enzymes
could get priority in further research for an enhanced ethanol
fermentation efficiency [128].
Acknowledgement
We are thankful to the University of Malaya for financial support of
this work under the project UM.C/HIR/MOHE/ENG/20.
References
[1] Vohra M, Manwar J, Manmode R, Padgilwar S, Patil S. Bioethanol production:
feedstock and current technologies. J Environ Chem Eng 2014;2:573–84.
[2] Vanhala P, Bergström I, Haaspuro T, Kortelainen P, Holmberg M, Forsius M.
Boreal forests can have a remarkable role in reducing greenhouse gas emissions
locally: land use-related and anthropogenic greenhouse gas emissions and sinks at
the municipal level. Sci Total Environ 2016;557:51–7.
[3] Kundu A, Sahu JN, Redzwan G, Hashim M. An overview of cathode material and
catalysts suitable for generating hydrogen in microbial electrolysis cell. Int J
Hydrog Energy 2013;38:1745–57.
[4] Abnisa F, Arami-Niya A, Daud WW, Sahu J. Characterization of bio-oil and bio-
char from pyrolysis of palm oil wastes. BioEnergy Res 2013;6:830–40.
[5] RFA. Industry statistics, Renewable Fuels Association, Washington, DC., USA.
〈http://ethanolrfa.org/resources/industry/statistics/#1454099788442-
e48b2782-ea53〉; 2016.
[6] Nigam PS, Singh A. Production of liquid biofuels from renewable resources. Prog
Energy Combust Sci 2011;37:52–68.
[7] Naik S, Goud VV, Rout PK, Dalai AK. Production of first and second generation
biofuels: a comprehensive review. Renew Sustain Energy Rev 2010;14:578–97.
[8] Bayraktar H. Experimental and theoretical investigation of using gasoline–ethanol
blends in spark-ignition engines. Renew Energy 2005;30:1733–47.
[9] Al-Hasan M. Effect of ethanol–unleaded gasoline blends on engine performance
and exhaust emission. Energy Convers Manag 2003;44:1547–61.
[10] Hara T, Tanoue K. Laminar flame speed of ethanol, n-heptane, iso-octane air
mixtures. JSAE paper; 20068518; 2006.
[11] Lynd LR. Overview and evaluation of fuel ethanol from cellulosic biomass:
technology, economics, the environment, and policy. Annu Rev Energy Environ
1996;21:403–65.
[12] Kar Y, Deveci H. Importance of P-series fuels for flexible-fuel vehicles (FFVs) and
alternative fuels. Energy Sources Part A 2006;28:909–21.
[13] Pickett J, Anderson D, Bowles D, Bridgwater T, Jarvis P, Mortimer N, et al.
Sustainable biofuels: prospects and challenges. London, UK: The Royal Society;
2008.
[14] McCarthy JE, Tiemann M. MTBE in gasoline: clean air and drinking water issues.
Washington, DC, USA; 1998.
[15] Green KR, Lowenbach WA. MTBE contamination: environmental, legal, and
public policy challengesguest editorial. Environ Forensics 2001;2:3–6.
[16] Yao C, Yang X, Roy Raine R, Cheng C, Tian Z, Li Y. The effects of MTBE/ethanol
additives on toxic species concentration in gasoline flame. Energy Fuels
2009;23:3543–8.
[17] Nwufo O, Nwafor O, Igbokwe J. Effects of blends on the physical properties of
bioethanol produced from selected Nigerian crops. Int J Ambient Energy
2016;37:10–5.
[18] Ho DP, Ngo HH, Guo W. A mini review on renewable sources for biofuel.
Bioresour Technol 2014;169:742–9.
[19] Sindhu R, Gnansounou E, Binod P, Pandey A. Bioconversion of sugarcane crop
residue for value added products–An overview. Renewable Energy; doi:http://dx.
doi.org/10.1016/j.renene.2016.02.057; 2016.
[20] Fish WW, Bruton BD, Russo VM. Watermelon juice: a promising feedstock
supplement, diluent, and nitrogen supplement for ethanol biofuel production.
Biotechnol Biofuels 2009;2:1–9.
[21] Louhichi B, Belgaib J, Benamor H, Hajji N. Production of bio-ethanol from three
varieties of dates. Renew Energy 2013;51:170–4.
[22] Soam S, Kumar R, Gupta RP, Sharma PK, Tuli DK, Das B. Life cycle assessment of
fuel ethanol from sugarcane molasses in northern and western India and its
impact on Indian biofuel programme. Energy 2015;83:307–15.
[23] Costa OYA, Souto BM, Tupinambá DD, Bergmann JC, Kyaw CM, Kruger RH, et al.
Microbial diversity in sugarcane ethanol production in a Brazilian distillery using a
H. Zabed et al.
culture-independent method. J Ind Microbiol Biotechnol 2015;42:73–84.
[24] Daniel PH, Lueschen WE, Kanne BK, Hoverstad TR. A comparison of sweet
sorghum cultivars and maize for ethanol production. J Prod Agric 1991;4:377–81.
[25] Laopaiboon L, Thanonkeo P, Jaisil P, Laopaiboon P. Ethanol production from
sweet sorghum juice in batch and fed-batch fermentations by Saccharomyces
cerevisiae. World J Microbiol Biotechnol 2007;23:1497–501.
[26] Marx S, Brandling J, van der Gryp P. Ethanol production from tropical sugar beet
juice. Afr J Biotechnol 2016;11:11709–20.
[27] Chandrasekaran M, Bahkali AH. Valorization of date palm (Phoenix dactylifera)
fruit processing by-products and wastes using bioprocess technology–Review.
Saudi J Biol Sci 2013;20:105–20.
[28] Chniti S, Djelal H, Hassouna M, Amrane A. Residue of dates from the food
industry as a new cheap feedstock for ethanol production. Biomass- Bioenergy
2014;69:66–70.
[29] Upadhyay A, Lama JP, Tawata S. Utilization of pineapple waste: a review. J Food
Sci Technol Nepal 2013;6:10–8.
[30] Ban-Koffi L, Han Y. Alcohol production from pineapple waste. World J Microbiol
Biotechnol 1990;6:281–4.
[31] Rodríguez L, Toro M, Vazquez F, Correa-Daneri M, Gouiric S, Vallejo M.
Bioethanol production from grape and sugar beet pomaces by solid-state
fermentation. Int J Hydrog Energy 2010;35:5914–7.
[32] Santana ÁL, MAA Meireles. New starches are the trend for industry applications: a
review. Food Public Health 2014;4:229–41.
[33] Balat M. Production of bioethanol from lignocellulosic materials via the bio-
chemical pathway: a review. Energy Convers Manag 2011;52:858–75.
[34] Kim S, Dale BE. Global potential bioethanol production from wasted crops and
crop residues. Biomass- Bioenergy 2004;26:361–75.
[35] Zabed H, Faruq G, Sahu J, Boyce A, Ganesan P. A comparative study on normal
and high sugary corn genotypes for evaluating enzyme consumption during dry-
grind ethanol production. Chem Eng J 2016;287:691–703.
[36] Murphy JD, Power NM. How can we improve the energy balance of ethanol
production from wheat?. Fuel 2008;87:1799–806.
[37] Pervez S, Aman A, Iqbal S, Siddiqui NN, Qader SAU. Saccharification and
liquefaction of cassava starch: an alternative source for the production of
bioethanol using amylolytic enzymes by double fermentation process. BMC
Biotechnol 2014;14:1.
[38] Huang Y, Jin Y, Fang Y, Li Y, Zhao H. Simultaneous utilization of non-starch
polysaccharides and starch and viscosity reduction for bioethanol fermentation
from fresh Canna edulis Ker. tubers. Bioresour Technol 2013;128:560–4.
[39] Huang Y, Jin Y, Fang Y, Li Y, Zhang G, Xiao Y, et al. Simultaneous saccharification
and fermentation (SSF) of non-starch polysaccharides and starch from fresh tuber
of Canna edulis ker at a high solid content for ethanol production. Biomass-
Bioenergy 2013;52:8–14.
[40] Ramírez MB, Ferrari MD, Lareo C. Fuel ethanol production from commercial
grain sorghum cultivars with different tannin content. J Cereal Sci
2016;69:125–31.
[41] George NA, Pecota KV, Bowen BD, Schultheis JR, Yencho GC. Root piece planting
in sweetpotato—A synthesis of previous research and directions for the future.
HortTechnology 2011;21:703–11.
[42] Duvernay WH, Chinn MS, Yencho GC. Hydrolysis and fermentation of sweet-
potatoes for production of fermentable sugars and ethanol. Ind Crops Prod
2013;42:527–37.
[43] Ziska LH, Runion GB, Tomecek M, Prior SA, Torbet HA, Sicher R. An evaluation
of cassava, sweet potato and field corn as potential carbohydrate sources for
bioethanol production in Alabama and Maryland. Biomass- Bioenergy
2009;33:1503–8.
[44] Khan RA, Nawaz A, Ahmed M, Khan MR, Azam F, Ullah S, et al. Production of
bioethanol through enzymatic hydrolysis of potato. Afr J Biotechnol
2014;11:6739–43.
[45] Thatoi H, Dash PK, Mohapatra S, Swain MR. Bioethanol production from tuber
crops using fermentation technology: a review. Int J Sustain Energy 2014:1–26.
[46] Susanto H. Fuel grade bioethanol production from Iles-iles
(Amorphophaluscampanulatus) tuber. Procedia Environ Sci 2015;23:199–206.
[47] Velásquez-Arredondo H, Ruiz-Colorado A, De Oliveira S. Ethanol production
process from banana fruit and its lignocellulosic residues: energy analysis. Energy
2010;35:3081–7.
[48] RFA . Ethanol industry outlook. Washington, DC., USA: Renewable Fuels
Association; 2015.
[49] Schnepf R, Yacobucci BD. Renewable Fuel Standard (RFS): overview and issues.
CRS Report for Congress; 2010.
[50] O'Hair SK. Tropical root and tuber crops. Hortic Rev 1990;12:157–96.
[51] Kays SJ, Wang Y. Sweetpotato quality: Its importance, assessment and selection in
breeding programs. I International Conference on Sweetpotato Food and Health
for the Future 583. p. 187-193; 2001.
[52] Scott G, Maldonado L. Sweetpotato for the new millennium: trends in production
and utilization in developing countries. CIP Program Report 1997;98:329–35.
[53] Saini JK, Saini R, Tewari L. Lignocellulosic agriculture wastes as biomass
feedstocks for second-generation bioethanol production: concepts and recent
developments. 3 Biotech; 2014:1–17.
[54] Kuhad RC, Singh A. Lignocellulose biotechnology: current and future prospects.
Crit Rev Biotechnol 1993;13:151–72.
[55] Lewandowski I, Scurlock JM, Lindvall E, Christou M. The development and
current status of perennial rhizomatous grasses as energy crops in the US and
Europe. Biomass- Bioenergy 2003;25:335–61.
[56] van der Weijde T, Kamei CLA, Torres AF, Vermerris W, Dolstra O, Visser RG, et al.
The potential of C4 grasses for cellulosic biofuel production. Front Plant Sci
2013:4.
[57] Rengsirikul K, Ishii Y, Kangvansaichol K, Sripichitt P, Punsuvon V, Vaithanomsat
P, et al. Biomass yield, chemical composition and potential ethanol yields of 8
cultivars of napiergrass (Pennisetum purpureum Schumach.) harvested 3-monthly
in central Thailand. J Sustain Bioenergy Syst 2013;3:107.
497
Renewable and Sustainable Energy Reviews 71 (2017) 475–501
[58] Kumar A, Singh L, Ghosh S. Bioconversion of lignocellulosic fraction of water-
hyacinth (Eichhornia crassipes) hemicellulose acid hydrolysate to ethanol by
Pichia stipitis. Bioresour Technol 2009;100:3293–7.
[59] Aswathy U, Sukumaran RK, Devi GL, Rajasree K, Singhania RR, Pandey A. Bio-
ethanol from water hyacinth biomass: an evaluation of enzymatic saccharification
strategy. Bioresour Technol 2010;101:925–30.
[60] Limayem A, Ricke SC. Lignocellulosic biomass for bioethanol production: current
perspectives, potential issues and future prospects. Prog Energy Combust Sci
2012;38:449–67.
[61] Lal R. World crop residues production and implications of its use as a biofuel.
Environ Int 2005;31:575–84.
[62] Kreith F, Krumdieck S. Principles of sustainable energy systems. CRC Press; 2013.
[63] Zhu J, Pan X. Woody biomass pretreatment for cellulosic ethanol production:
technology and energy consumption evaluation. Bioresour Technol
2010;101:4992–5002.
[64] Chen Y. Evaluating woody biomass for ethanol production. Biomass- Prod Technol
2011:3–7.
[65] Zhu J, Luo X, Tian S, Gleisner R, Negrone J, Horn E. Efficient ethanol production
from beetle-killed lodgepole pine using SPORL technology and Saccharomyces
cerevisiae without detoxification. TAPPI J 2011;10:9–18.
[66] Gonzalez RW, Treasure T, Phillips RB, Jameel H, Saloni D. Economics of cellulosic
ethanol production: green liquor pretreatment for softwood and hardwood,
greenfield and repurpose scenarios. Bioresources 2011;6:2551–67.
[67] Buah W, Cunliffe A, Williams P. Characterization of products from the pyrolysis of
municipal solid waste. Process Saf Environ Prot 2007;85:450–7.
[68] Prasad S, Singh A, Joshi H. Ethanol as an alternative fuel from agricultural,
industrial and urban residues. Resour Conserv Recycl 2007;50:1–39.
[69] Li S, Zhang X, Andresen JM. Production of fermentable sugars from enzymatic
hydrolysis of pretreated municipal solid waste after autoclave process. Fuel
2012;92:84–8.
[70] Sánchez C. Lignocellulosic residues: biodegradation and bioconversion by fungi.
Biotechnol Adv 2009;27:185–94.
[71] Perlack RD, Wright LL, Turhollow AF, Graham RL, Stokes BJ, Erbach DC.
Biomass as feedstock for a bioenergy and bioproducts industry: the technical
feasibility of a billion-ton annual supply. DTIC Document; 2005.
[72] Hadar Y. Sources for lignocellulosic raw materials for the production of ethanol.
Lignocellulose Conversion: Springer; 2013:21–38.
[73] Quintero J, Montoya M, Sánchez O, Giraldo O, Cardona C. Fuel ethanol
production from sugarcane and corn: comparative analysis for a Colombian case.
Energy 2008;33:385–99.
[74] Goldemberg J, Guardabassi P. The potential for first‐generation ethanol produc-
tion from sugarcane. Biofuels Bioprod Bioref 2010;4:17–24.
[75] Sathitsuksanoh N, Zhu Z, Zhang Y-HP. Cellulose solvent-and organic solvent-
based lignocellulose fractionation enabled efficient sugar release from a variety of
lignocellulosic feedstocks. Bioresour Technol 2012;117:228–33.
[76] Maluf WR. Proving Interdisciplinary Education in BiofuelsTechnology. 1st Brazil-
US Biofuels Short Course. Universidade de São Paulo, Brazil. 〈http://www.iea.usp.
br/midiateca/apresentacao/malufbiofuels.pdf〉; 2009.
[77] Bali G, Ramamurthi R, Sullia SB, Kasturi S. EnvironBiotechnology. New Delhi,
India: A P H Publishing Corporation; 2002.
[78] Badger P. Trends in new crops and new uses. Alexandria VA: ASHS Press; 2002.
[79] Nasidi M, Agu R, Deeni Y, Walker G. Fermentation of stalk juices from different
Nigerian sorghum cultivars to ethanol. Bioethanol 2013:1.
[80] Zhao YL, Dolat A, Steinberger Y, Wang X, Osman A, Xie GH. Biomass yield and
changes in chemical composition of sweet sorghum cultivars grown for biofuel.
Field Crops Res 2009;111:55–64.
[81] McLeod J, May W, Salmon D, Sosulski K, Thomas J, Brown P, et al. Changes in
ethanol production potential due to species, cultivar and location on the Canadian
prairie. Can J Plant Sci 2010;90:163–71.
[82] Lareo C, Ferrari MD, Guigou M, Fajardo L, Larnaudie V, Ramírez MB, et al.
Evaluation of sweet potato for fuel bioethanol production: hydrolysis and
fermentation. SpringerPlus 2013;2:1.
[83] Barros‐Rios J, Romaní A, Garrote G, Ordas B. Biomass, sugar, and bioethanol
potential of sweet corn. GCB Bioenergy 2015;7:153–60.
[84] Dhaliwal SS, Oberoi HS, Sandhu SK, Nanda D, Kumar D, Uppal SK. Enhanced
ethanol production from sugarcane juice by galactose adaptation of a newly
isolated thermotolerant strain of Pichiakudriavzevii. Bioresour Technol
2011;102:5968–75.
[85] Mamma D, Christakopoulos P, Koullas D, Kekos D, Macris B, Koukios E. An
alternative approach to the bioconversion of sweet sorghum carbohydrates to
ethanol. Biomass- Bioenergy 1995;8:99–103.
[86] Ratnavathi C, Chakravarthy SK, Komala V, Chavan U, Patil J. Sweet sorghum as
feedstock for biofuel production: a review. Sugar Tech 2011;13:399–407.
[87] Razmovski R, Vučurović V. Ethanol production from sugar beet molasses by S.
cerevisiae entrapped in an alginate–maize stem ground tissue matrix. Enzym
Microb Technol 2011;48:378–85.
[88] Zheng Y, Lee C, Yu C, Cheng Y-S, Simmons CW, Zhang R, et al. Ensilage and
bioconversion of grape pomace into fuel ethanol. J Agric Food Chem
2012;60:11128–34.
[89] Corbin KR, Hsieh YS, Betts NS, Byrt CS, Henderson M, Stork J, et al. Grape marc
as a source of carbohydrates for bioethanol: chemical composition, pre-treatment
and saccharification. Bioresour Technol 2015;193:76–83.
[90] Gullón B, Falqué E, Alonso JL, Parajó JC. Evaluation of apple pomace as a raw
material for alternative applications in food industries. Food Technol Biotechnol
2007;45:426–33.
[91] Al-Shahib W, Marshall RJ. The fruit of the date palm: its possible use as the best
food for the future?. Int J Food Sci Nutr 2003;54:247–59.
[92] Behera S, Mohanty RC, Ray RC. Comparative study of bio-ethanol production
from mahula (Madhuca latifolia L.) flowers by Saccharomyces cerevisiae and
Zymomonas mobilis. Appl Energy 2010;87:2352–5.
[93] Tomaszewska M, Białończyk L. Ethanol production from whey in a bioreactor
H. Zabed et al.
coupled with direct contact membrane distillation. Catal Today 2016.
[94] Bothast R, Schlicher M. Biotechnological processes for conversion of corn into
ethanol. Appl Microbiol Biotechnol 2005;67:19–25.
[95] Lang X, Macdonald D, Hill G. Recycle bioreactor for bioethanol production from
wheat starch II. fermentation and economics. Energy Sources 2001;23:427–36.
[96] Rani P, Sharma S, Garg F, Raj K, Wati L. Ethanol production from potato flour by
Saccharomyces cerevisiae. Indian J Sci Technol 2010;3:733–6.
[97] Ferrari MD, Guigou M, Lareo C. Energy consumption evaluation of fuel bioethanol
production from sweet potato. Bioresour Technol 2013;136:377–84.
[98] Vijaya Sarathi Reddy O, Basappa S. Preparation of sweet potato flour and its
fermentation to ethanol. J Food Sci Technol 1997;34:108–12.
[99] Griffey C, Brooks W, Kurantz M, Thomason W, Taylor F, Obert D, et al. Grain
composition of Virginia winter barley and implications for use in feed, food, and
biofuels production. J Cereal Sci 2010;51:41–9.
[100] Thomas K, Dhas A, Rossnagel B, Ingledew W. Production of fuel alcohol from
hull-less barley by very high gravity technology. Cereal Chem 1995;72:360–4.
[101] Nichols NN, Dien BS, Wu YV, Cotta MA. Ethanol fermentation of starch from field
peas. Cereal Chem 2005;82:554–8.
[102] Kuiper L, Ekmekci B, Hamelinck C, Hettinga W, Meyer S, Koop K. Bio-ethanol
from cassava. Ecofys Neth Bv 2007:1–3.
[103] Egbebi AOBT. Chemical compositions of ripe and unripe banana and plaintain.
Int J Trop Med Public Health 2012;1:1–5.
[104] Lebot V. Tropical root and tuber crops: cassava, sweet potato, yams and aroids:
Cabi; 2009.
[105] Long X-H, Shao H-B, Liu L, Liu L-P, Liu Z-P. Jerusalem artichoke: a sustainable
biomass feedstock for biorefinery. Renew Sustain Energy Rev 2016;54:1382–8.
[106] Cinelli BA, López JA, Castilho LR, Freire DM, Castro AM. Granular starch
hydrolysis of babassu agroindustrial residue: a bioprocess within the context of
biorefinery. Fuel 2014;124:41–8.
[107] Nwokocha LM, Williams PA. Comparative study of physicochemical properties of
breadfruit (Artocarpus altilis) and white yam starches. Carbohydr Polym
2011;85:294–302.
[108] Li M, Liu P, Zou W, Yu L, Xie F, Pu H, et al. Extrusion processing and
characterization of edible starch films with different amylose contents. J Food Eng
2011;106:95–101.
[109] Cruz BR, Abraão AS, Lemos AM, Nunes FM. Chemical composition and functional
properties of native chestnut starch (Castanea sativa Mill). Carbohydr Polym
2013;94:594–602.
[110] Braga ME, Moreschi SR, Meireles MAA. Effects of supercritical fluid extraction on
Curcuma longa L. and Zingiber officinale R. starches. Carbohydr Polym
2006;63:340–6.
[111] Van Hung P, Morita N. Chemical compositions, fine structure and physicochem-
ical properties of kudzu (Pueraria lobata) starches from different regions. Food
Chem 2007;105:749–55.
[112] Sotomayor C, Frias J, Fornal J, Sadowska J, Urbano G, Vidal-Valverde C. Lentil
starch content and its microscopical structure as influenced by natural fermen-
tation. Starch-Starke 1999;51:152–6.
[113] Zhu F. Structure, physicochemical properties, and uses of millet starch. Food Res
Int 2014;64:200–11.
[114] Albano KM, Franco CM, Telis VR. Rheological behavior of Peruvian carrot starch
gels as affected by temperature and concentration. Food Hydrocoll
2014;40:30–43.
[115] Kadam S, Salunkhe D, Maga JA. Nutritional composition, processing, and
utilization of horse gram and moth bean. critical Reviews in food Science & .
Nutrition 1985;22:1–26.
[116] McKendry P. Energy production from biomass (part 1): overview of biomass.
Bioresour Technol 2002;83:37–46.
[117] Brosse N, Hage RE, Sannigrahi P, Ragauskas A. Dilute sulphuric acid and ethanol
organosolv pretreatment of Miscanthus x Giganteus. Cellul Chem Technol
2010;44:71–8.
[118] Saxena R, Adhikari D, Goyal H. Biomass-based energy fuel through biochemical
routes: a review. Renew Sustain Energy Rev 2009;13:167–78.
[119] Howard R, Abotsi E, Van Rensburg EJ, Howard S. Lignocellulose biotechnology:
issues of bioconversion and enzyme production. Afr J Biotechnol 2004;2:602–19.
[120] Zhu Y, Lee Y, Elander RT. Optimization of dilute-acid pretreatment of corn stover
using a high-solids percolation reactorTwenty-Sixth Symposium on Biotechnology
for Fuels and Chemicals. Springer; 2005. p. 1045–54.
[121] Kim M, Day DF. Composition of sugar cane, energy cane, and sweet sorghum
suitable for ethanol production at Louisiana sugar mills. J Ind Microbiol
Biotechnol 2011;38:803–7.
[122] Ludueña L, Fasce D, Alvarez VA, Stefani PM. Nanocellulose from rice husk
following alkaline treatment to remove silica. Bioresources 2011;6:1440–53.
[123] Miao M, Li R, Huang C, Jiang B, Zhang T. Impact of β-amylase degradation on
properties of sugary maize soluble starch particles. Food Chem 2015;177:1–7.
[124] Singh V, Graeber J. Effect of corn hybrid variability and planting location on dry
grind ethanol production. Trans Asae 2005;48:709–14.
[125] Reicks G, Woodard HJ, Bly A. Improving the fermentation characteristics of corn
through agronomic and processing practices. Agron J 2009;101:201–6.
[126] Copeland L, Blazek J, Salman H, Tang MC. Form and functionality of starch. Food
Hydrocoll 2009;23:1527–34.
[127] Yangcheng H, Jiang H, Blanco M, Jane J-l. Characterization of normal and waxy
corn starch for bioethanol production. J Agric Food Chem 2013;61:379–86.
[128] Gírio F, Fonseca C, Carvalheiro F, Duarte L, Marques S, Bogel-Łukasik R.
Hemicelluloses for fuel ethanol: a review. Bioresour Technol 2010;101:4775–800.
[129] Zabed H, Sahu J, Boyce A, Faruq G. Fuel ethanol production from lignocellulosic
biomass: an overview on feedstocks and technological approaches. Renew Sustain
Energy Rev 2016;66:751–74.
[130] Chang VS, Holtzapple MT. Fundamental factors affecting biomass enzymatic
reactivity. Twenty-first symposium on biotechnology for fuels and chemicals.
Springer; 2000:5–37.
[131] Mosier N, Wyman C, Dale B, Elander R, Lee Y, Holtzapple M, et al. Features of
498
Renewable and Sustainable Energy Reviews 71 (2017) 475–501
promising technologies for pretreatment of lignocellulosic biomass. Bioresour
Technol 2005;96:673–86.
[132] Drummond A-RF, Drummond IW. Pyrolysis of sugar cane bagasse in a wire-mesh
reactor. Ind Eng Chem Res 1996;35:1263–8.
[133] de Souza RB, de Menezes JAS, de Souza RdFR, Dutra ED, de Morais MA. Jr,
Mineral Composition of the Sugarcane Juice and Its Influence on the Ethanol
Fermentation. Appl Biochem Biotechnol 2015;175:209–22.
[134] Kawa-Rygielska J, Pietrzak W, Regiec P, Stencel P. Utilization of concentrate after
membrane filtration of sugar beet thin juice for ethanol production. Bioresour
Technol 2013;133:134–41.
[135] Ratnavathi C, Suresh K, Kumar BV, Pallavi M, Komala V, Seetharama N. Study on
genotypic variation for ethanol production from sweet sorghum juice. Biomass-
Bioenergy 2010;34:947–52.
[136] Soontornchaiboon W, Chunhachart O, Pawongrat R. Ethanol production from
Yam Bean using yeast Saccharomyces cerevisiae TISTR 5339. In: Proceedings of
the 1st Mae Fah Luang University International Conference Northern Thailand;
2012.
[137] Bouallagui H, Touhami Y, Hanafi N, Ghariani A, Hamdi M. Performances
comparison between three technologies for continuous ethanol production from
molasses. Biomass- Bioenergy 2013;48:25–32.
[138] Dionisi FAR, Aoki IV, Calabrese RJSugar cane juice clarification process. U.S.
Patent No. 7,338,562. 4 Mar. 2008. 2008.
[139] de la Piscina PR, Homs N. Use of biofuels to produce hydrogen (reformation
processes). Chem Soc Rev 2008;37:2459–67.
[140] Barcelos CA, Maeda RN, Betancur GJV, Pereira N. Jr., Ethanol production from
sorghum grains [Sorghum bicolor (L.) Moench]: evaluation of the enzymatic
hydrolysis and the hydrolysate fermentability. Braz J Chem Eng
2011;28:597–604.
[141] Mussatto SI, Dragone G, Guimarães PM, Silva JPA, Carneiro LM, Roberto IC,
et al. Technological trends, global market, and challenges of bio-ethanol produc-
tion. Biotechnol Adv 2010;28:817–30.
[142] Johnston DB, McAloon AJ. Protease increases fermentation rate and ethanol yield
in dry-grind ethanol production. Bioresour Technol 2014;154:18–25.
[143] Shapouri H, Duffield JA, Wang MQ. The energy balance of corn ethanol: an
update. United States Department of Agriculture, Economic Research Service;
2002.
[144] Mueller S. National dry mill corn ethanol survey. Biotechnol Lett
2008;2010(32):1261–4.
[145] Bothast RJ. New technologies in biofuel production. Agric Outlook Forum 2005:6.
[146] Mosier NS, Ileleji KE. How fuel ethanol is made from corn. Bioenergy: Biomass-
Biofuels 2014:379.
[147] Kim Y, Mosier N, Ladisch MR. Process simulation of modified dry grind ethanol
plant with recycle of pretreated and enzymatically hydrolyzed distillers' grains.
Bioresour Technol 2008;99:5177–92.
[148] Erickson G, Klopfenstein T, Adams D, Rasby R. General overview of feeding corn
milling co-products to beef cattle. Linclon, NE, USA: Corn processing co-products
manual: University of Nebraska; 2005.
[149] Zabed H, Boyce AN, Sahu JN, Faruq G. Evaluation of the quality of dried distiller's
grains with solubles for normal and high sugary corn genotypes during dry–grind
ethanol production. J Clean Prod 2016;14:4282–93.
[150] Zabed H, Boyce A, Faruq G, Sahu J. A comparative evaluation of agronomic
performance and kernel composition of normal and high sugary corn genotypes
(Zea mays L.) grown for dry-grind ethanol production. Ind Crops Prod
2016;94:9–19.
[151] Zabed H, Faruq G, Boyce AN, Sahu JN, Ganesan P. Evaluation of high sugar
containing corn genotypes as viable feedstocks for decreasing enzyme consump-
tion during dry-grind ethanol production. J Taiwan Inst Chem Eng
2016;58:467–75.
[152] Shi A, Du Z, Ma X, Cheng Y, Min M, Deng S, et al. Production and evaluation of
biodiesel and bioethanol from high oil corn using three processing routes.
Bioresour Technol 2013;128:100–6.
[153] Hao X, Thelen K, Gao J. Prediction of the ethanol yield of dry-grind maize grain
using near infrared spectroscopy. Biosyst Eng 2012;112:161–70.
[154] Białas W, Szymanowska D, Grajek W. Fuel ethanol production from granular corn
starch using Saccharomyces cerevisiae in a long term repeated SSF process with
full stillage recycling. Bioresour Technol 2010;101:3126–31.
[155] Nkomba EY, van Rensburg E, Chimphango AF, Görgens JF. The influence of
sorghum grain decortication on bioethanol production and quality of the distillers’
dried grains with solubles using cold and conventional warm starch processing.
Bioresour Technol 2016;203:181–9.
[156] Lee W-S, Chen I-C, Chang C-H, Yang S-S. Bioethanol production from sweet
potato by co-immobilization of saccharolytic molds and Saccharomyces cerevi-
siae. Renew Energy 2012;39:216–22.
[157] Schweinberger CM, Putti TR, Susin GB, Trierweiler JO, Trierweiler LF. Ethanol
production from sweet potato: the effect of ripening, comparison of two heating
methods, and cost analysis. Can J Chem Eng 2016.
[158] Montesinos T, Navarro J-M. Production of alcohol from raw wheat flour by
Amyloglucosidase and Saccharomyces cerevisiae. Enzym Microb Technol
2000;27:362–70.
[159] Vu VH, Kim K. Ethanol production from rice winery waste-rice wine cake by
simultaneous saccharification and fermentation without cooking. J Microbiol
Biotechnol 2009;19:1161–8.
[160] Aimaretti NR, Ybalo CV, Rojas ML, Plou FJ, Yori JC. Production of bioethanol
from carrot discards. Bioresour Technol 2012;123:727–32.
[161] Reddy L, Reddy O. Improvement of ethanol production in very high gravity
fermentation by horse gram (Dolichos biflorus) flour supplementation. Lett Appl
Microbiol 2005;41:440–4.
[162] Wan C, Li Y. Microbial pretreatment of corn stover with Ceriporiopsis sub-
vermispora for enzymatic hydrolysis and ethanol production. Bioresour Technol
2010;101:6398–403.
[163] Cai D, Dong Z, Wang Y, Chen C, Li P, Qin P, et al. Biorefinery of corn cob for
H. Zabed et al.
microbial lipid and bio-ethanol production: an environmental friendly process.
Bioresour Technol 2016;211:677–84.
[164] Saha BC, Nichols NN, Qureshi N, Kennedy GJ, Iten LB, Cotta MA. Pilot scale
conversion of wheat straw to ethanol via simultaneous saccharification and
fermentation. Bioresour Technol 2015;175:17–22.
[165] Wi SG, Choi IS, Kim KH, Kim HM, Bae H-J. Bioethanol production from rice
straw by popping pretreatment. Biotechnol Biofuels 2013;6:1.
[166] Ramos LP, da Silva L, Ballem AC, Pitarelo AP, Chiarello LM, Silveira MHL.
Enzymatic hydrolysis of steam-exploded sugarcane bagasse using high total solids
and low enzyme loadings. Bioresour Technol 2015;175:195–202.
[167] Santos J, Lucena M, Gusmão N, Gouveia E. Optimization of ethanol production by
Saccharomyces cerevisiae UFPEDA 1238 in simultaneous saccharification and
fermentation of delignified sugarcane bagasse. Ind Crops Prod 2012;36:584–8.
[168] Marx S, Ndaba B, Chiyanzu I, Schabort C. Fuel ethanol production from sweet
sorghum bagasse using microwave irradiation. Biomass- Bioenergy
2014;65:145–50.
[169] Han M, Kang KE, Kim Y, Choi G-W. High efficiency bioethanol production from
barley straw using a continuous pretreatment reactor. Process Biochem
2013;48:488–95.
[170] Karunanithy C, Muthukumarappan K, Gibbons W. Extrusion pretreatment of pine
wood chips. Appl Biochem Biotechnol 2012;167:81–99.
[171] Kim HY, Hong C-Y, Kim S-H, Yeo H, Choi I-G. Optimization of the organosolv
pretreatment of yellow poplar for bioethanol production by response surface
methodology. Mokchae Konghak=J Korean Wood Sci Technol 2015;43:600–12.
[172] Ahn DJ, Kim SK, Yun HS. Optimization of pretreatment and saccharification for
the production of bioethanol from water hyacinth by Saccharomyces cerevisiae.
Bioprocess Biosyst Eng 2012;35:35–41.
[173] Capecchi L, Galbe M, Wallberg O, Mattarelli P, Barbanti L. Combined ethanol and
methane production from switchgrass (Panicum virgatum L.) impregnated with
lime prior to steam explosion. Biomass- Bioenergy 2016;90:22–31.
[174] Njoku SI, Iversen JA, Uellendahl H, Ahring BK. Production of ethanol from
hemicellulose fraction of cocksfoot grass using pichia stipitis. Sustain Chem
Process 2013;1:13.
[175] Li A, Antizar-Ladislao B, Khraisheh M. Bioconversion of municipal solid waste to
glucose for bio-ethanol production. Bioprocess Biosyst Eng 2007;30:189–96.
[176] Wyman CE, Dale BE, Elander RT, Holtzapple M, Ladisch MR, Lee Y. Coordinated
development of leading biomass pretreatment technologies. Bioresour Technol
2005;96:1959–66.
[177] Sørensen A, Teller PJ, Hilstrøm T, Ahring BK. Hydrolysis of Miscanthus for
bioethanol production using dilute acid presoaking combined with wet explosion
pre-treatment and enzymatic treatment. Bioresour Technol 2008;99:6602–7.
[178] Kamzon MA, Abderafi S, Bounahmidi T. Promising bioethanol processes for
developing a biorefinery in the Moroccan sugar industry. Int J Hydrog Energy
2016;41:20880–96.
[179] Szczodrak J, Fiedurek J. Technology for conversion of lignocellulosic biomass to
ethanol. Biomass- Bioenergy 1996;10:367–75.
[180] Sarkar N, Ghosh SK, Bannerjee S, Aikat K. Bioethanol production from agricul-
tural wastes: an overview. Renew Energy 2012;37:19–27.
[181] Cavka A, Jönsson LJ. Detoxification of lignocellulosic hydrolysates using sodium
borohydride. Bioresour Technol 2013;136:368–76.
[182] Palmqvist E, Hahn-Hägerdal B. Fermentation of lignocellulosic hydrolysates. II:
inhibitors and mechanisms of inhibition. Bioresour Technol 2000;74:25–33.
[183] Larsson S, Reimann A, Nilvebrant N-O, Jönsson LJ. Comparison of different
methods for the detoxification of lignocellulose hydrolyzates of spruce. Appl
Biochem Biotechnol 1999;77:91–103.
[184] Klinke HB, Thomsen A, Ahring BK. Inhibition of ethanol-producing yeast and
bacteria by degradation products produced during pre-treatment of biomass. Appl
Microbiol Biotechnol 2004;66:10–26.
[185] Chandel AK, Da Silva SS, Singh OV. Detoxification of lignocellulose hydrolysates:
biochemical and metabolic engineering toward white biotechnology. BioEnergy
Res 2013;6:388–401.
[186] Guo M, Song W, Buhain J. Bioenergy and biofuels: history, status, and
perspective. Renew Sustain Energy Rev 2015;42:712–25.
[187] Taherzadeh MJ, Karimi K. Acid-based hydrolysis processes for ethanol from
lignocellulosic materials: a review. BioResources 2007;2:472–99.
[188] Liu Z, Ho S-H, Hasunuma T, Chang J-S, Ren N-Q, Kondo A. Recent advances in
yeast cell-surface display technologies for waste biorefineries. Bioresour Technol
2016. http://dx.doi.org/10.1016/j.biortech.2016.03.132.
[189] Farone WA, Cuzens JE. Method of producing sugars using strong acid hydrolysis
of cellulosic and hemicellulosic materials. Google Patents; 1996.
[190] Yang L, Liu Y, Ruan R, Wang Y, Zeng W, Liu C, et al. Advances in production of 5-
hydroxymethylfurfural from starch. Mod Chem Ind 2011;1:014.
[191] Kim K, Hamdy M. Acid hydrolysis of sweet potato for ethanol production.
Biotechnol Bioeng 1985;27:316–20.
[192] Buckow R, Weiss U, Heinz V, Knorr D. Stability and catalytic activity of α‐amylase
from barley malt at different pressure–temperature conditions. Biotechnol Bioeng
2007;97:1–11.
[193] Lamsal B, Wang H, Johnson L. Effect of corn preparation methods on dry-grind
ethanol production by granular starch hydrolysis and partitioning of spent beer
solids. Bioresour Technol 2011;102:6680–6.
[194] Plumier BM, Danao M-GC, Rausch KD, Singh V. Changes in unreacted starch
content in corn during storage. J Stored Prod Res 2015;61:85–9.
[195] Sanchez OJ, Cardona CA. Trends in biotechnological production of fuel ethanol
from different feedstocks. Bioresour Technol 2008;99:5270–95.
[196] Rakin M, Mojovic L, Nikolic S, Vukasinovic M, Nedovic V. Bioethanol production
by immobilized Sacharomyces cerevisiae var. ellipsoideus cells. Afr J Biotechnol
2009:8.
[197] Lamsal B, Yoo J, Brijwani K, Alavi S. Extrusion as a thermo-mechanical pre-
treatment for lignocellulosic ethanol. Biomass- Bioenergy 2010;34:1703–10.
[198] Lavarack B, Griffin G, Rodman D. The acid hydrolysis of sugarcane bagasse
hemicellulose to produce xylose, arabinose, glucose and other products. Biomass-
499
Renewable and Sustainable Energy Reviews 71 (2017) 475–501
Bioenergy 2002;23:367–80.
[199] Chandel AK, Chan E, Rudravaram R, Narasu ML, Rao LV, Ravindra P. Economics
and environmental impact of bioethanol production technologies: an appraisal.
Biotechnol Mol Biol Rev 2007;2:14–32.
[200] Wyman CE. Biomass ethanol: technical progress, opportunities, and commercial
challenges. Annu Rev Energy Environ 1999;24:189–226.
[201] Lee Y, Iyer P, Torget RW. Dilute-acid hydrolysis of lignocellulosic biomass. Recent
progress in bioconversion of lignocellulosics: Springer; 1999:93-115.
[202] Fengel D, Wegener G. Wood: chemistry, ultrastructure, reactions. Walter de
Gruyter; 1983.
[203] Balat M. An overview of biofuels and policies in the European Union. Energy
Sources, Part B 2007;2:167–81.
[204] Zhang Y-HP, Ding S-Y, Mielenz JR, Cui J-B, Elander RT, Laser M, et al.
Fractionating recalcitrant lignocellulose at modest reaction conditions. Biotechnol
Bioeng 2007;97:214–23.
[205] Duff SJ, Murray WD. Bioconversion of forest products industry waste cellulosics
to fuel ethanol: a review. Bioresour Technol 1996;55:1–33.
[206] Reczey K, Szengyel Z, Eklund R, Zacchi G. Cellulase production by T. reesei.
Bioresour Technol 1996;57:25–30.
[207] Lee J. Biological conversion of lignocellulosic biomass to ethanol. J Biotechnol
1997;56:1–24.
[208] Bajaj BK, Pangotra H, Wani MA, Sharma P, Sharma A. Partial purification and
characterization of a highly thermostable and pH stable endoglucanase from a
newly isolated Bacillus strain M-9. Indian J Chem Technol 2009;16:382–7.
[209] Saha BC. Lignocellulose biodegradation and applications in biotechnology. ACS
symposium series: Washington, DC; American Chemical Society; 1999; 2004:2–
35.
[210] Saha BC. Hemicellulose bioconversion. J Ind Microbiol Biotechnol
2003;30:279–91.
[211] Sarris D, Papanikolaou S. Biotechnological production of ethanol: biochemistry,
processes and technologies. Eng Life Sci 2016. http://dx.doi.org/10.1002/
elsc.201400199.
[212] Singh V, Rausch KD, Yang P, Shapouri H, Belyea RL, Tumbleson ME. Modified
dry grind ethanol process. University of Illinois atUrbana–Champaign Report No
UILU; 2001.
[213] Boulton RB, Singleton VL, Bisson LF, Kunkee RE. Yeast and biochemistry of
ethanol fermentation. Principles and Practices of Winemaking. Springer; 1999. p.
102–92.
[214] Wingren A, Galbe M, Zacchi G. Techno‐economic evaluation of producing ethanol
from softwood: Comparison of SSF and SHF and identification of bottlenecks.
Biotechnol Progress 2003;19:1109–17.
[215] Delgenes J, Moletta R, Navarro J. The effect of aeration on d-xylose fermentation
byPachysolen tannophilus, Pichia stipitis, Kluyveromyces marxianus andCandida
shehatae. Biotechnol Lett 1986;8:897–900.
[216] Slininger P, Branstrator L, Bothast R, Okos M, Ladisch M. Growth, death, and
oxygen uptake kinetics of Pichia stipitis on xylose. Biotechnol Bioeng
1991;37:973–80.
[217] Delgenes J, Moletta R, Navarro J. Fermentation of D‐xylose, D‐glucose, L‐
arabinose mixture by Pichia stipitis: Effect of the oxygen transfer rate on
fermentation performance. Biotechnol Bioeng 1989;34:398–402.
[218] Taniguchi M, Itaya T, Tohma T, Fujii M. Ethanol production from a mixture of
glucose and xylose by a novel co-culture system with two fermentors and two
microfiltration modules. J Ferment Bioeng 1997;84:59–64.
[219] Oliveira SC, Oliveira RC, Tacin MV, Gattás EA. Kinetic Modeling and optimization
of a batch ethanol fermentation process. J Bioprocess Biotech 2016:2016.
[220] Thomas K, Hynes S, Ingledew W. Practical and theoretical considerations in the
production of high concentrations of alcohol by fermentation. Process Biochem
1996;31:321–31.
[221] Olsson L, Hahn-Hägerdal B. Fermentation of lignocellulosic hydrolysates for
ethanol production. Enzym Microb Technol 1996;18:312–31.
[222] Gnansounou E, Dauriat A. Ethanol fuel from biomass: a review. J Sci Ind Res
2005;64:809–21.
[223] Roukas T. Ethanol production from non-sterilized beet molasses by free and
immobilized Saccharomyces cerevisiae cells using fed-batch culture. J Food Eng
1996;27:87–96.
[224] Brethauer S, Wyman CE. Review: continuous hydrolysis and fermentation for
cellulosic ethanol production. Bioresour Technol 2010;101:4862–74.
[225] Dien B, Cotta M, Jeffries T. Bacteria engineered for fuel ethanol production:
current status. Appl Microbiol Biotechnol 2003;63:258–66.
[226] Chen Y, Sheng J, Jiang T, Stevens J, Feng X, Wei N. Transcriptional profiling
reveals molecular basis and novel genetic targets for improved resistance to
multiple fermentation inhibitors in Saccharomyces cerevisiae. Biotechnol Biofuels
2016;9:1.
[227] Snoek T, Verstrepen KJ, Voordeckers K. How do yeast cells become tolerant to
high ethanol concentrations?. Curr Genet 2016:1–6.
[228] Kosaric N, Velikonja J. Liquid and gaseous fuels from biotechnology: challenge
and opportunities. FEMS Microbiol Rev 1995;16:111–42.
[229] Lin Y, Tanaka S. Ethanol fermentation from biomass resources: current state and
prospects. Appl Microbiol Biotechnol 2006;69:627–42.
[230] Bai F, Anderson W, Moo-Young M. Ethanol fermentation technologies from sugar
and starch feedstocks. Biotechnol Adv 2008;26:89–105.
[231] Cazetta M, Celligoi M, Buzato J, Scarmino I. Fermentation of molasses by
Zymomonas mobilis: effects of temperature and sugar concentration on ethanol
production. Bioresour Technol 2007;98:2824–8.
[232] Rogers P, Lee K, Skotnicki M, Tribe D. Ethanol production by Zymomonas
mobilisMicrobial reactions. Springer; 1982. p. 37–84.
[233] Parker C, Peekhaus N, Zhang X, Conway T. Kinetics of sugar transport and
phosphorylation influence glucose and fructose cometabolism by Zymomonas
mobilis. Appl Environ Microbiol 1997;63:3519–25.
[234] Swings J, De Ley J. The biology of Zymomonas. Bacteriol Rev 1977;41:1.
[235] Sprenger GA. Carbohydrate metabolism in Zymomonas mobilis: a catabolic
H. Zabed et al.
highway with some scenic routes. FEMS Microbiol Lett 1996;145:301–7.
[236] Lee S-W, Ebata T, Liu Y-C, Tanaka H. Co-immobilization of three strains of
microorganisms and its application in ethanol production from raw starch under
unsterile conditions. J Ferment Bioeng 1993;75:36–42.
[237] Lim LH, Macdonald DG, Hill GA. Hydrolysis of starch particles using immobilized
barley α-amylase. Biochem Eng J 2003;13:53–62.
[238] Hahn-Hägerdal B, Galbe M, Gorwa-Grauslund M-F, Lidén G, Zacchi G. Bio-
ethanol–the fuel of tomorrow from the residues of today. Trends Biotechnol
2006;24:549–56.
[239] Liao JC, Mi L, Pontrelli S, Luo S. Fuelling the future: microbial engineering for the
production of sustainable biofuels. Nat Rev Microbiol 2016.
[240] Hahn-Hägerdal B, Karhumaa K, Jeppsson M, Gorwa-Grauslund MF. Metabolic
engineering for pentose utilization in Saccharomyces cerevisiae. Biofuels.
Springer; 2007. p. 147–77.
[241] Jeffries TW. Engineering the Pichia stipitis genome for fermentation of hemi-
cellulose hydrolysates. Washington, DC: Bioenergy ASM Press; 2008. p. 37–47.
[242] Kuyper M, Toirkens MJ, Diderich JA, Winkler AA, Dijken JP, Pronk JT.
Evolutionary engineering of mixed sugar utilization by a xylose fermenting
Saccharomyces cerevisiae strain. FEMS yeast Res 2005;5:925–34.
[243] Wisselink HW, Toirkens MJ, Wu Q, Pronk JT, van Maris AJ. Novel evolutionary
engineering approach for accelerated utilization of glucose, xylose, and arabinose
mixtures by engineered Saccharomyces cerevisiae strains. Appl Environ Microbiol
2009;75:907–14.
[244] Sonderegger M, Schümperli M, Sauer U. Metabolic engineering of a phosphoke-
tolase pathway for pentose catabolism in Saccharomyces cerevisiae. Appl Environ
Microbiol 2004;70:2892–7.
[245] Aydemir E, Demirci S, Doğan A, Aytekin A, Sahin F. Genetic Modifications of
Saccharomyces cerevisiae for Ethanol Production from Starch Fermentation: a
Review. J Bioprocess Biotech 2014;4:2.
[246] Murai T, Ueda M, Yamamura M, Atomi H, Shibasaki Y, Kamasawa N, et al.
Construction of a starch-utilizing yeast by cell surface engineering. Appl Environ
Microbiol 1997;63:1362–6.
[247] Shigechi H, Fujita Y, Koh J, Ueda M, Fukuda H, Kondo A. Energy-saving direct
ethanol production from low-temperature-cooked corn starch using a cell-surface
engineered yeast strain co-displaying glucoamylase and α-amylase. Biochem Eng
J 2004;18:149–53.
[248] Shigechi H, Uyama K, Fujita Y, Matsumoto T, Ueda M, Tanaka A, et al. Efficient
ethanol production from starch through development of novel flocculent yeast
strains displaying glucoamylase and co-displaying or secreting α-amylase. J Mol
Catal B: Enzym 2002;17:179–87.
[249] Ohta K, Beall D, Mejia J, Shanmugam K, Ingram L. Genetic improvement of
Escherichia coli for ethanol production: chromosomal integration of Zymomonas
mobilis genes encoding pyruvate decarboxylase and alcohol dehydrogenase II.
Appl Environ Microbiol 1991;57:893–900.
[250] Wood BE, Ingram L. Ethanol production from cellobiose, amorphous cellulose,
and crystalline cellulose by recombinant Klebsiella oxytoca containing chromo-
somally integrated Zymomonas mobilis genes for ethanol production and
plasmids expressing thermostable cellulase genes from Clostridium thermocel-
lum. Appl Environ Microbiol 1992;58:2103–10.
[251] Lim Y, Jang Y, Kim K. Production of a high concentration of ethanol from potato
tuber by high gravity fermentation. Food Sci Biotechnol 2013;22:441–8.
[252] Nikolić S, Mojović L, Rakin M, Pejin D, Pejin J. Ultrasound-assisted production of
bioethanol by simultaneous saccharification and fermentation of corn meal. Food
Chem 2010;122:216–22.
[253] Tomás-Pejó E, Oliva J, González A, Ballesteros I, Ballesteros M. Bioethanol
production from wheat straw by the thermotolerant yeast Kluyveromyces marx-
ianus CECT 10875 in a simultaneous saccharification and fermentation fed-batch
process. Fuel 2009;88:2142–7.
[254] Erdei B, Galbe M, Zacchi G. Simultaneous saccharification and co-fermentation of
whole wheat in integrated ethanol production. Biomass- Bioenergy
2013;56:506–14.
[255] Srichuwong S, Fujiwara M, Wang X, Seyama T, Shiroma R, Arakane M, et al.
Simultaneous saccharification and fermentation (SSF) of very high gravity (VHG)
potato mash for the production of ethanol. Biomass- Bioenergy 2009;33:890–8.
[256] Laopaiboon L, Nuanpeng S, Srinophakun P, Klanrit P, Laopaiboon P. Ethanol
production from sweet sorghum juice using very high gravity technology: effects of
carbon and nitrogen supplementations. Bioresour Technol 2009;100:4176–82.
[257] Masiero SS, Peretti A, Trierweiler LF, Trierweiler JO. Simultaneous cold hydro-
lysis and fermentation of fresh sweet potato. Biomass- Bioenergy
2014;70:174–83.
[258] Nikolić S, Mojović L, Rakin M, Pejin D. Bioethanol production from corn meal by
simultaneous enzymatic saccharification and fermentation with immobilized cells
of Saccharomyces cerevisiae var. ellipsoideus. Fuel 2009;88:1602–7.
[259] Liang L, Zhang Y-p , Zhang L, Zhu M-j, Liang S-z, Huang Y-n. Study of sugarcane
pieces as yeast supports for ethanol production from sugarcane juice and
molasses. J Ind Microbiol Biotechnol 2008;35:1605–13.
[260] Zhao J, Xia L. Ethanol production from corn stover hemicellulosic hydrolysate
using immobilized recombinant yeast cells. Biochem Eng J 2010;49:28–32.
[261] Li S, Li G, Zhang L, Zhou Z, Han B, Hou W, et al. A demonstration study of
ethanol production from sweet sorghum stems with advanced solid state
fermentation technology. Appl Energy 2013;102:260–5.
[262] Horisawa S, Ando H, Ariga O, Sakuma Y. Direct ethanol production from
cellulosic materials by consolidated biological processing using the wood rot
fungus Schizophyllum commune. Bioresour Technol 2015;197:37–41.
[263] Chu-Ky S, Pham T-H, Bui K-LT, Nguyen T-T, Pham K-D, Nguyen H-DT, et al.
Simultaneous liquefaction, saccharification and fermentation at very high gravity
of rice at pilot scale for potable ethanol production and distillers dried grains
composition. Food Bioprod Process 2016;98:79–85.
[264] Szymanowska D, Grajek W. Energy-saving and by-products-free production of
ethanol from granular corn starch. Biotechnol J Biotechnol Comput Biol
Bionanotechnol 2011;92.
500
Renewable and Sustainable Energy Reviews 71 (2017) 475–501
[265] Szymanowska-Powałowska D, Lewandowicz G, Kubiak P, Błaszczak W. Stability of
the process of simultaneous saccharification and fermentation of corn flour. The
effect of structural changes of starch by stillage recycling and scaling up of the
process. Fuel 2014;119:328–34.
[266] Białas W, Czerniak A, Szymanowska-Powałowska D. Kinetic modeling of simul-
taneous saccharification and fermentation of corn starch for ethanol production.
Acta Biochim Pol 2014;61:153–62.
[267] Mojović L, Nikolić S, Rakin M, Vukasinović M. Production of bioethanol from corn
meal hydrolyzates. Fuel 2006;85:1750–5.
[268] Robertson GH, Wong DW, Lee CC, Wagschal K, Smith MR, Orts WJ. Native or
raw starch digestion: a key step in energy efficient biorefining of grain. J Agric
Food Chem 2006;54:353–65.
[269] Uthumporn U, Zaidul IS, Karim A. Hydrolysis of granular starch at sub-
gelatinization temperature using a mixture of amylolytic enzymes. Food Bioprod
Process 2010;88:47–54.
[270] Foerster H. Granular starch hydrolysis (GSHE) for conversion of grains to ethanol.
Near-term Opportunities for Biorefineries Symposium. Champaign, IL: Genencor-
A Danisco Division; 2010.
[271] Szymanowska-Powałowska D, Lewandowicz G, Błaszczak W, Szwengiel A.
Structural changes of corn starch during fuel ethanol production from corn flour.
BioTechnologia 2012;93:333–41.
[272] Srichuwong S. Jane J-l. methods for characterization of residual starch in
distiller's dried grains with solubles (DDGS). Cereal Chem 2011;88:278–82.
[273] Wang P, Singh V, Xu L, Johnston DB, Rausch KD, Tumbleson M. Comparison of
enzymatic (E-Mill) and conventional dry-grind corn processes using a granular
starch hydrolyzing enzyme. Cereal Chem 2005;82:734–8.
[274] Li J, Vasanthan T, Bressler DC. Improved cold starch hydrolysis with urea
addition and heat treatment at subgelatinization temperature. Carbohydr Polym
2012;87:1649–56.
[275] Genencor. STARGEN™ 002: Granular starch hydrolyzing enzyme for ethanol
production; 2009.
[276] Duan G, Dunn-Coleman N, Lantero O, Pilgrim CE, Shetty JK. Acid fungal protease
in fermentation of insoluble starch substrates. Google Patents; 2009.
[277] Gohel V, Duan G, Maisuria V. Impact of an acid fungal protease in high gravity
fermentation for ethanol production using indian sorghum as a feedstock.
Biotechnol Prog 2013;29:329–36.
[278] Nikolić S, Mojović L, Pejin D, Rakin M, Vukašinović M. Production of bioethanol
from corn meal hydrolyzates by free and immobilized cells of Saccharomyces
cerevisiae var. ellipsoideus. Biomass- Bioenergy 2010;34:1449–56.
[279] Mishra A, Sharma AK, Sharma S, Bagai R, Mathur AS, Gupta RP, et al.
Lignocellulosic ethanol production employing immobilized Saccharomyces cere-
visiae in packed bed reactor. Renew Energy 2016.
[280] Kourkoutas Y, Bekatorou A, Banat IM, Marchant R, Koutinas A. Immobilization
technologies and support materials suitable in alcohol beverages production: a
review. Food Microbiol 2004;21:377–97.
[281] Zabed H, Faruq G, Sahu JN, Azirun MS, Hashim R, Nasrulhaq Boyce A.
Bioethanol production from fermentable sugar juice. Sci World J 2014:2014.
[282] Najafpour G, Younesi H, Ismail KSK. Ethanol fermentation in an immobilized cell
reactor using Saccharomyces cerevisiae. Bioresour Technol 2004;92:251–60.
[283] Stratford M. Evidence for two mechanisms of flocculation in Saccharomyces
cerevisiae. Yeast 1989;5:S441–S445.
[284] Maiorella B, Blanch H, Wilke C, Wyman CE. Economic evaluation of alternative
ethanol fermentation processes. Biotechnol Bioenergy 2009;104:419–43.
[285] Alkasrawi M, Galbe M, Zacchi G. Recirculation of process streams in fuel ethanol
production from softwood based on simultaneous saccharification and fermenta-
tion. Biotechnology for Fuels and Chemicals, Springer; 2002. p. 849–61.
[286] Bayrock D, Ingledew WM. Application of multistage continuous fermentation for
production of fuel alcohol by very-high-gravity fermentation technology. J Ind
Microbiol Biotechnol 2001;27:87–93.
[287] Ingledew W, Thomas K, Hynes S, McLeod J. Viscosity concerns with rye mashes
used for ethanol production. Cereal Chem 1999;76:459–64.
[288] Jeong Y-J, Seo J-H, Lee J-B, Jang S-M, Shin S-R, Kim K-S. Changes in the
components during alcohol fermentation of potatoes using pilot system. Korean J
Food Preserv 2000;7:233–9.
[289] Tasić MB, Konstantinović BV, Lazić ML, Veljković VB. The acid hydrolysis of
potato tuber mash in bioethanol production. Biochem Eng J 2009;43:208–11.
[290] Casey GP, Ingledew WM. Ethanol tolerance in yeasts. CRC Crit Rev Microbiol
2008.
[291] Kawa-Rygielska J, Pietrzak W. Ethanol fermentation of very high gravity (VHG)
maize mashes by Saccharomyces cerevisiae with spent brewer's yeast supple-
mentation. Biomass- Bioenergy 2014;60:50–7.
[292] Pradeep P, Reddy O. High gravity fermentation of sugarcane molasses to produce
ethanol: effect of nutrients. Indian J Microbiol 2010;50:82–7.
[293] Chang J-W, Lin Y-H, Huang L-Y, Duan K-J. The effect of fermentation config-
urations and fan supplementation on ethanol production from sorghum grains
under very-high-gravity conditions. J Taiwan Inst Chem Eng 2011;42:1–4.
[294] Xiao D, Wu S, Zhu X, Chen Y, Guo X. Effects of soya fatty acids on cassava ethanol
fermentation. Appl Biochem Biotechnol 2010;160:410–20.
[295] Sankh SN, Deshpande PS, Arvindekar AU. Improvement of ethanol production
using Saccharomyces cerevisiae by enhancement of biomass and nutrient
supplementation. Appl Biochem Biotechnol 2011;164:1237–45.
[296] Yu J, Tan T. Ethanol production by solid state fermentation of sweet sorghum
using thermotolerant yeast strain. Fuel Process Technol 2008;89:1056–9.
[297] Wang E-Q, Li S-Z, Tao L, Geng X, Li T-C. Modeling of rotating drum bioreactor for
anaerobic solid-state fermentation. Appl Energy 2010;87:2839–45.
[298] Han B, Wang L, Li S, Wang E, Zhang L, Li T. Ethanol production from sweet
sorghum stalks by advanced solid state fermentation (ASSF) technologyEthanol
production from sweet sorghum stalks by advanced solid state fermentation
(ASSF) technology]. Sheng wu gong cheng xue bao=Chin J Biotechnol
2010;26:966–73.
[299] Lynd LR, Laser MS, Bransby D, Dale BE, Davison B, Hamilton R, et al. How
H. Zabed et al.
biotech can transform biofuels. Nat Biotechnol 2008;26:169–72.
[300] Margeot A, Hahn-Hagerdal B, Edlund M, Slade R, Monot F. New improvements
for lignocellulosic ethanol. Curr Opin Biotechnol 2009;20:372–80.
[301] Hamelinck CN, Van Hooijdonk G, Faaij AP. Ethanol from lignocellulosic biomass:
techno-economic performance in short-, middle-and long-term. Biomass-
Bioenergy 2005;28:384–410.
[302] Borzani W, Jurkiewicz C. Variation of the ethanol yield during very rapid batch
fermentation of sugar-cane blackstrap molasses. Braz J Chem Eng
1998;15:225–33.
[303] Ramchandran D, Johnston DB, Tumbleson M, Rausch KD, Singh V. Seasonal
variability in ethanol concentrations from a dry grind fermentation operation
associated with incoming corn variability. Ind Crops Prod 2015;67:155–60.
[304] Li Z, Liu W, Gu Z, Li C, Hong Y, Cheng L. The effect of starch concentration on the
gelatinization and liquefaction of corn starch. Food Hydrocoll 2015;48:189–96.
[305] Imtiaz U, Assadzadeh A, Jamuar SS, Sahu J. Bioreactor temperature profile
controller using inverse neural network (INN) for production of ethanol. J Process
Control 2013;23:731–42.
[306] Ballesteros M, Oliva J, Negro M, Manzanares P, Ballesteros I. Ethanol from
lignocellulosic materials by a simultaneous saccharification and fermentation
process (SFS) with < i > Kluyveromyces marxianus < i > CECT 10875. Process
Biochem 2004;39:1843–8.
[307] Phisalaphong M, Srirattana N, Tanthapanichakoon W. Mathematical modeling to
investigate temperature effect on kinetic parameters of ethanol fermentation.
Biochem Eng J 2006;28:36–43.
[308] Torija MJ, Rozes N, Poblet M, Guillamón JM, Mas A. Effects of fermentation
temperature on the strain population of Saccharomyces cerevisiae. Int J Food
Microbiol 2003;80:47–53.
[309] Liu R, Shen F. Impacts of main factors on bioethanol fermentation from stalk juice
of sweet sorghum by immobilized Saccharomyces cerevisiae (CICC 1308).
Bioresour Technol 2008;99:847–54.
[310] Lyness E, Doelle HW. Fermentation pattern of sucrose to ethanol conversions by
Zymomonas mobilis. Biotechnol Bioeng 1981;23:1449–60.
[311] Lee K, Skotnicki M, Tribe D, Rogers P. The effect of temperature on the kinetics of
ethanol production by strains of Zymomonas mobilis. Biotechnol Lett
1981;3:291–6.
[312] Banat IM, Nigam P, Marchant R. Isolation of thermotolerant, fermentative yeasts
growing at 52C and producing ethanol at 45C and 50C. World J Microbiol
Biotechnol 1992;8:259–63.
[313] Abdel-Banat BM, Hoshida H, Ano A, Nonklang S, Akada R. High-temperature
fermentation: how can processes for ethanol production at high temperatures
become superior to the traditional process using mesophilic yeast?. Appl
Microbiol Biotechnol 2010;85:861–7.
[314] McMeekin T, Olley J, Ratkowsky D, Ross T. Predictive microbiology: towards the
interface and beyond. Int J Food Microbiol 2002;73:395–407.
[315] Gao C, Fleet G. The effects of temperature and pH on the ethanol tolerance of the
wine yeasts, Saccharomyces cerevisiae, Candida stellata and Kloeckera apiculata. J
Appl Bacteriol 1988;65:405–9.
[316] Carlsen M, Nielsen J, Villadsen J. Kinetic studies of acid-inactivation of α-amylase
from Aspergillus oryzae. Chem Eng Sci 1996;51:37–43.
[317] Kasemets K, Nisamedtinov I, Laht T-M, Abner K, Paalme T. Growth character-
istics of Saccharomyces cerevisiae S288C in changing environmental conditions:
auxo-accelerostat study. Antonie Van Leeuwenhoek 2007;92:109–28.
[318] Lin Y, Zhang W, Li C, Sakakibara K, Tanaka S, Kong H. Factors affecting ethanol
fermentation using Saccharomyces cerevisiae BY4742. Biomass- Bioenergy
2012;47:395–401.
[319] Onsoy T, Thanonkeo P, Thanonkeo S, Yamada M. Ethanol production from
Jerusalem artichoke by Zymomonas mobilis in batch fermentation. KMITL Sci
Technol J 2007;7:55–60.
[320] Hill G, Macdonald D, Lang X. a-amylase inhibition and inactivation in barley malt
during cold starch hydrolysis. Biotechnol Lett 1997;19:1139–41.
[321] Ingledew W. Alcohol production by Saccharomyces cerevisiae: a yeast primer.
Chapter 5. In: Jacques KA, Lyons TP, Kelsall DR, editors. The alcohol textbook.
Nottingham, UK: Nottingham University Press; 1998.
[322] Modenbach AA, Nokes SE. Enzymatic hydrolysis of biomass at high-solids
loadings–a review. Biomass- Bioenergy 2013;56:526–44.
[323] Zabed H, Sahu J, Faruq G, Ganesan P, Boyce AChanges in biomass composition,
enzymatic hydrolysis and calculated ethanol yields with genotypic variation of
corn. Engineering and Technology (BICET 2014), In: Proceedings of the 5th
501
Renewable and Sustainable Energy Reviews 71 (2017) 475–501
Brunei International Conference on: IET; 2014. p.1-5.
[324] Szymanowska-Powalowska D, Lewandowicz G, Blaszczak W, Szwengiel A.
Structural changes of corn starch during fuel ethanol production from corn flour.
Biotechnol J Biotechnol Comput Biol Bionanotechnol 2012:93.
[325] Takeshige K, Ouchi K. Effects of yeast invertase on ethanol production in
molasses. J Ferment Bioeng 1995;79:513–5.
[326] Lemuz CR, Dien BS, Singh V, McKinney J, Tumbleson M, Rausch KD.
Development of an ethanol yield procedure for dry-grind corn processing. Cereal
Chem 2009;86:355–60.
[327] Breisha GZ. Production of 16% ethanol from 35% sucrose. Biomass- Bioenergy
2010;34:1243–9.
[328] Thomas K, Hynes S, Ingledew W. Effect of lactobacilli on yeast growth, viability
and batch and semi‐continuous alcoholic fermentation of corn mash. J Appl
Microbiol 2001;90:819–28.
[329] Skinner KA, Leathers TD. Bacterial contaminants of fuel ethanol production. J Ind
Microbiol Biotechnol 2004;31:401–8.
[330] Narendranath N, Hynes S, Thomas K, Ingledew W. Effects of lactobacilli on yeast-
catalyzed ethanol fermentations. Appl Environ Microbiol 1997;63:4158–63.
[331] Narendranath NV, Power R. Relationship between pH and medium dissolved
solids in terms of growth and metabolism of Lactobacilli and Saccharomyces
cerevisiae during ethanol production. Appl Environ Microbiol 2005;71:2239–43.
[332] Graves T, Narendranath NV, Dawson K, Power R. Effect of pH and lactic or acetic
acid on ethanol productivity by Saccharomyces cerevisiae in corn mash. J Ind
Microbiol Biotechnol 2006;33:469–74.
[333] Sarris D, Papanikolaou S. Biotechnological production of ethanol: biochemistry,
processes and technologies. Eng Life Sci 2016.
[334] Murthy GS, Singh V, Johnston DB, Rausch KD, Tumbleson M. Improvement in
fermentation characteristics of degermed ground corn by lipid supplementation. J
Ind Microbiol Biotechnol 2006;33:655–60.
[335] Gnansounou E, Dauriat A, Villegas J, Panichelli L. Life cycle assessment of
biofuels: energy and greenhouse gas balances. Bioresour Technol
2009;100:4919–30.
[336] Gerbrandt K, Chu PL, Simmonds A, Mullins KA, MacLean HL, Griffin WM, et al.
Life cycle assessment of lignocellulosic ethanol: a review of key factors and
methods affecting calculated GHG emissions and energy use. Curr Opin
Biotechnol 2016;38:63–70.
[337] Gallejones P, Pardo G, Aizpurua A, Del Prado A. Life cycle assessment of first-
generation biofuels using a nitrogen crop model. Sci Total Environ
2015;505:1191–201.
[338] Daylan B, Ciliz N. Life cycle assessment and environmental life cycle costing
analysis of lignocellulosic bioethanol as an alternative transportation fuel. Renew
Energy 2016;89:578–87.
[339] Amores MJ, Mele FD, Jiménez L, Castells F. Life cycle assessment of fuel ethanol
from sugarcane in Argentina. Int J Life Cycle Assess 2013;18:1344–57.
[340] Kim S, Dale BE. Life cycle assessment of fuel ethanol derived from corn grain via
dry milling. Bioresour Technol 2008;99:5250–60.
[341] Kumar D, Murthy GS. Life cycle assessment of energy and GHG emissions during
ethanol production from grass straws using various pretreatment processes. Int J
Life Cycle Assess 2012;17:388–401.
[342] Morales M, Quintero J, Conejeros R, Aroca G. Life cycle assessment of
lignocellulosic bioethanol: environmental impacts and energy balance. Renew
Sustain Energy Rev 2015;42:1349–61.
[343] da Silva ARG, Ortega CET, Rong B-G. Techno-economic analysis of different
pretreatment processes for lignocellulosic-based bioethanol production. Bioresour
Technol 2016;218:561–70.
[344] Chen H, Venditti R, Gonzalez R, Phillips R, Jameel H, Park S. Economic
evaluation of the conversion of industrial paper sludge to ethanol. Energy Econ
2014;44:281–90.
[345] McAloon A, Taylor F, Yee W, Ibsen K, Wooley R. Determining the cost of
producing ethanol from corn starch and lignocellulosic feedstocks [Technical
Report NREL/TP-580-28893]. Golden, CO (USA): National Renewable Energy
Laboratory; 2000.
[346] Gregg D, Boussaid A, Saddler J. Techno-economic evaluations of a generic wood-
to-ethanol process: effect of increased cellulose yields and enzyme recycle.
Bioresour Technol 1998;63:7–12.
[347] Della-Bianca BE, Basso TO, Stambuk BU, Basso LC, Gombert AK. What do we
know about the yeast strains from the Brazilian fuel ethanol industry?. Appl
Microbiol Biotechnol 2013;97:979–91.