Review - Bioethanol Production From Tuber Crop Using Fermentation Technology

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Bioethanol production from tuber

crops using fermentation technology: a
review
a a a
Hrudayanath Thatoi , Preeti Krishna Dash , Sonali Mohapatra &
b
Manas Ranjan Swain
a
Department of Biotechnology, College of Engineering
and Technology, Biju Pattnaik University of Technology,
Bhubaneswar-751003, India
b
Department of Biotechnology, Indian Institute of Technology
Click for updates Madras, Chennai-600036, India
Published online: 27 Jun 2014.
To cite this article: Hrudayanath Thatoi, Preeti Krishna Dash, Sonali Mohapatra & Manas Ranjan
Swain (2014): Bioethanol production from tuber crops using fermentation technology: a review,
International Journal of Sustainable Energy, DOI: 10.1080/14786451.2014.918616
To link to this article: http://dx.doi.org/10.1080/14786451.2014.918616
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International Journal of Sustainable Energy, 2014
http://dx.doi.org/10.1080/14786451.2014.918616
Bioethanol production from tuber crops using fermentation

technology: a review
Hrudayanath Thatoia∗ , Preeti Krishna Dasha , Sonali Mohapatraa and Manas Ranjan Swainb
a Department of Biotechnology, College of Engineering and Technology, Biju Pattnaik University of
Technology, Bhubaneswar-751003, India; b Department of Biotechnology, Indian Institute of Technology
Madras, Chennai-600036, India
(Received 6 December 2013; final version received 19 April 2014)
Bioethanol, an alcohol produced by fermentation of plant biomass containing starch and sugars by micro-
organisms, considered as a dominant form of fuel for future. Production of this renewable fuel, especially
from starchy materials such as tuber crops, holds a remarkable potential to meet the future energy demand
because of its high production and comparitively less demand for use as food and fodder. This review focuses
on the world bioethanol production scenario from various tuber crops, namely cassava, sweet potato, potato,
yam, aroids, sugar beet, etc., fermentation techniques and micro-organisms used in fermentation process
along with its future prospects. The advances in metabolic pathway engineering and genetic engineering
techniques have led to the development of micro-organisms capable of efficiently converting biomass
sugars into ethanol. Several biotechnological tools that are also available for the improvement of microor-
ganisms to meet the harsh environments typically met with certain industrial fermentation process are also
discussed.
Keywords: tuber crops; micro-organisms; fermentation techniques; bioethanol; biotechnological tools
1. Introduction
In recent years, the annual energy consumption from petroleum sources has increased many-
fold resulting not only in continuous depletion of limited fossil fuel stocks but also a cause of
concern for the safer, better and greener environment (Lynd and Wang 2003). Further, the high
prices of fossil fuels have led to an energy crisis in both developing and developed countries
that are oil dependent. Reserve of fossil fuels is going to be depleted fast leading to the increase
in fuel prices and simultaneously unfolding energy crisis. Naylor et al. (2007) stated that ‘bio-
fuels will remain critical energy development target in many parts, if petroleum prices exceed
US$ 55–60 per barrel’. Therefore, there is a considerable emphasis on the development of bio-
fuel production technologies from plant sources and bioethanol production from plant biomass.
Bioethanol is an alcohol produced by fermentation of plant biomass, containing starch or sugars
and its production depends upon feedstock availability, variability and sustainability (Liimatainen,
Kuokkanen, and Kaarianen 2004).
Bioethanol is produced by fermentation of sugar by micro-organisms, as opposed to syn-
thetically produced ethanol from petrochemical sources. It is produced by the process of
∗ Corresponding author. Email: hn_thatoi@rediffmail.com
© 2014 Taylor & Francis

2 H. Thatoi et al.
Table 1. Worldwide bioethanol production from different feedstocks.
Leading ethanol producing regions worldwide

Ethanol production (million litres)
Location Feedstock 2007 2008 2009 2010 2011 2012
The America (North, Corn, sugarcane, wheat, etc. 45,546 60,402 66,416 77,548 76,400 75,915
Central and South)
Europe Grains, sugarbeet, wheat, rye 1882 2855 3645 4254 4429 4973
Asia/Pacific Corn, cassava, sugarcane, etc 2142 2753 2927 3115 3520 3965
Africa Corn, sugarcane 55 65 100 130 150 235
Source: F.O. Litch: Cited in Ethanol producer magazine, 27 June 2012. Available at http://www.ethanolproducer.com/articles/8907
distillation of fermented sugars, which can be utilised as a liquid fuel in internal combustion
engines, either neet or in blends with petroleum (Graeme and Walker 2010). It is a cleaner
transportation fuel burning than non-renewable gasoline, as it is more oxygenated (Wheals

et al. 1999). Biomass-based ethanol or bioethanol is well entrenched in policy as a potential
substitute for petroleum-based gasoline. One of the primary benefits of switching to this fuel is
that biomass can be renewed and can potentially provide a sustainable fuel supply over a long-
term period (Farrell et al. 2006). As an alternative source of energy, bioethanol can meet the
fuel demand as well as can save the environment from pollution problem. There are two types
of ethanol: (1) denatured ethanol and (2) hydrous ethanol (MacLean and Lave 2004). Denatured
ethanol is the one that is used in blending Premium Motor Spirit to E10, E20, etc., but unfit for
human consumption. A 10% bioethanol blend can effectively replace methyl tertiary butyl ether
(MTBE) as an oxygenation additive to gasoline and potentially prevent MTBE associated health
risks. When bioethanol replaces aromatic and sulphur-containing compounds used in gasoline, it
may also reduce nitrogen oxide emissions to improve air quality, which can reduce urban smog
(O’Connor 1994). The high oxygen content in bioethanol could reduce the generation of known
hazardous volatile organic compounds and carbon monoxide in vehicle exhaust. It is usually mixed
with 2–5 low-cost hydrocarbon that makes it unfit for human consumption. On the other hand,
hydrous ethanol is the natural ethanol used as industrial raw material in the production of alcoholic
beverages, cosmetics, perfumes, drinks, medicaments and other industrial uses (O’Connor 1994).
The global biofuel production has grown from 16 billion litres in 2000 to about 105 billion litres
in 2010 (i.e. 82% bioethanol and 18% biodiesel), accounting for about 3% of today’s transport
fuel on an energy basis (IEA 2011). In recent years the global ethanol industry continues to be a
bright spot in the world economy. Global production of bioethanol showed an upward trend over
the last 25 years, with a sharp increase from 2000. The annual bioethanol production worldwide
was around 83.1 billion litres in 2012 (Ibrahim 2012). Table 1 provides data on region wise
bioethanol production worldwide from 2007 to 2012 (F.O. Licht 2012) which slightly differs
from the values cited in this text from other sources. Further the trend in world ethanol production
during the year 2007–2012 is given in Figure 1. A study on bioethanol production revealed that
the global leaders in bioethanol production are: USA with production reaching to ∼50 billion
litres (primarily from maize and secondarily from wheat) and Brazil with ∼35 billion litres (from
sugarcane) in 2012 (Walker 2010). Together, the USA and Brazil produce 87% of the world’s
ethanol (Biofuels factsheet, 2013).
Following Brazil are the European countries like France, Germany, Spain and Sweden, where
bioethanol production increased by 71% in 2007, which depicts an increase of 58% from the year
2006 (Balat and Balat 2009). Asia ranks fourth in the world and the largest producer of ethanol in
Asia is China, with a total alcohol production of approximately 3.65 and 3.80 billion litres in 2004
and 2005, respectively (Patni, Pillai, and Dwivedi 2011). The annual production of bioethanol in
China in 2012 was noted to be 2433 million litres, which was a rise of 8% in comparison to the
International Journal of Sustainable Energy 3
Figure 1. World bioethanol production.

Source: F.O. Litch: Cited in Ethanol producer magazine, 27 June 2012. Available at <http://www.ethanolproducer.
com/articles/8907>.
year 2010 (USDA GAIN, China Biofuel Annual 2012). Among other Asian countries, India is a
country with a positive outlook towards renewable energy technologies and is committed to the
use of renewable sources to supplement its energy requirements. In 2010, India registered an 85%
increase in bio-fuel production from the previous year. Though India holds only 0.3% share of
the global production in 2010 (Patni, Pillai, and Dwivedi 2011), yet it continues to be the second
largest producer of ethanol in Asia, with an annual production of 2170 million litres in the year
2012 (USDA GAIN, India Biofuel Annual 2012).
The whole scientific approaches for biofuel production can be broadly divided into four major
generations depending on the substrate utilisation, technology involvement and micro-organisms
involved in the process. First-generation bioethanol is produced from food crops (sugary and
starchy crops) through fermentation. Second-generation biofuels have been developed to over-
come the limitations of first-generation biofuels. These fuels are produced from non-food crops,
such as wood, organic waste, food crop waste and specific biomass crops. Cellulosic ethanol is the
most developed second-generation biofuel and is produced from the cellulose or cell wall of plant
cells. The drawbacks are that the feedstock costs remain high, which is often due to processing
(shredding, densifying, pulverising and handling) and transportation. Third-generation biofuels
are generally produced from algal biomass, which has a very distinctive growth yield as compared
with lignocellulosic biomass (Brennan and Owende 2010). Fourth-generation biofuels are helpful
not only for producing sustainable energy but also in capturing and storing CO2 . Biomass mate-
rials, which have absorbed CO2 while growing, are converted into fuel using the same processes
as second-generation biofuels. This process differs from second- and third-generation production
because in this, at all stages of production, the carbon dioxide is captured using processes such
as oxy-fuel combustion (Yoon et al. 2010). Bioethanol production from different generations of
feedstocks with examples is shown in Figure 2.
Recently first-generation biofuel has been in news as being the culprit behind rising food prices.
Although they may contribute to higher food prices, it is a very small effect, and the debate does
not consider the environmental and energy security benefits of biofuels. Root and tuber crops
are used as food for human beings and also an important source of animal feed and industrial
products. On a global basis, approximately 45% of produced root and tuber crops are consumed
as food, with the remainder used as animal feed or for industrial processing of products such
4 H. Thatoi et al.
Figure 2. Biomass for different generation bioethanol production.
as bioethanol. In developed countries, the consumption of foods from root and tuber crops is
considerably smaller than that in developing countries. Thus surplus tuber crops that are not used
for food can be utilised for production of fuels without affecting the food supply for the human
beings. Besides root crop production has significantly increased during the period 1980–2012, as a
result of increase in area under production, improved planting materials and technologies, among
others (Jata et al. 2012). In India, major tuber crop productions are cassava and sweet potato as
compared to yam and other tuber crops. It has been surveyed that 60% of cassava produced in
India is for industrial purpose, 30% for human consumption and 10% for animal feed (Srinivas
2007). Similarly, sweet potatoes are also one of the major crops that are being extensively studied
and tested for bioethanol production with an average yield of up to 30 tonnes/ha. For industrial
bioethanol production, sweet potatoes with higher content of starch and better agricultural yields
are selected and they do not compete with crop resources needed to feed people (Jata et al. 2012).
It is not intended to be used as a food crop because its appearance, colour and flavour are not
attractive to be used as food (Duvernay, Chinn, and Yencho 2013). Moreover, tuber crops are
mostly used by tribals and poor people as food while it is not a favourite food for others. The
survey clearly demonstrates that a major part of these tuber crops can be utilised for industrial
purposes like production of bioethanol, leaving a sufficient amount to be consumed as a food for
human beings and animals (Jata et al. 2012).
With high concentrations of starch, the tuber crops are considered one of the most important first-
generation feedstocks for bioethanol production. Though the first- and other-generation biofuels
have several advantages and disadvantages, still the first-generation biofuels are more desirable
than the others because of their availability with the recently developed technologies and also these
feedstock can be produced in decentralised facilities, which allow the farmers to sell their own final
products in the market and thus encourages them in their production. In comparison to the first
generation, the disadvantage with the other generation biofuels is, it is not yet confirmed, that the
other generation biofuels can demonstrate better economic and energy performances as compared
with the first-generation feedstock (Campbell, Lobell, and Field 2009). With the increased use
of tubers for ethanol production, it is anticipated that increased efficiency in production will be
brought to impact on agriculture and these advantages will then be of benefit (Wheals et al. 1999).
This review focuses on the economical and efficient process of bioethanol production from tuber
crops and the prospect of application of biotechnological tools for high ethanol production, which
would be considered as the next generation fuel for the industry.
2. Types of tuber crops
Roots and tubers contribute to the energy and nutrition requirements and are increasingly used
for industrial purposes (Scott et al. 2000). Most of these crops are grown in non-agricultural lands
and add to the income generation of rural poor people. Tuber crops are produced in plenty in
developing countries as primary crops (Nweke 2004). These are the second group of cultivated
species after cereals, contributing substantially to human and animal food apart from finding use
in various industrial applications, including bioethanol production.
The major tuber crops which are used in bioethanol production include cassava, sweet potato,
potato, yam, aroids, sugar beet, etc. Bioethanol production per hectare from various tuber crops
and sugary substrates are given in Table 2. Generally tuber crops contain fair amount of starch
(16–24%), which furnish them as a raw material for ethanol production (Hoover 2001). The tubers
accumulate high levels of polysaccharides (fructans) during their growth. On a dry weight basis, the
tubers contain 68–83% fructans, 15–16% proteins, 13% insoluble fibre and 5% of other substances
including minerals. The chemical constituents of the tuber crops are shown in Table 3. Tuber crops
might be a better option for bioethanol because these are cheap and generally not favoured as food
and not used for valuable product formation (Zhang et al. 2011). The study also indicates that the
root crops have greater potential for ethanol production than corn grains, provided economical
harvesting and processing techniques are developed (Wheals et al. 1999).The major tuber crops
that are grown in India and worldwide with their annual bioethanol production are given in Table 4.
Table 2. Bioethanol production potential of different tuber

crops and sugary substrates.
Tuber crops and Ethanol

sugary substrates production L/ha References
Cassava 4711 Drapoch (2008)

Sweet potato 4800 Lareo et al. (2013)
Potato 1800 Osuji et al. (2010)
Sugar beet 4200 Kymalainen (2007)
Yam 700 Osuji et al. (2010)
Aroids 200 Cheng (2010)
Corn 2430 Kraatz (2008)
Molasses 1950 MINO (2010)
Sugar cane 6500 Cheng et al. (2011)
Table 3. Chemical composition of different tuber crops used in bioethanol production.
Tuber Moisture Protein Crude Nitrogen free Vitamin Starch

crop (g %) (g %) (g %) extract (g %) C (mg/g) (%)
Cassava 59.4 0.7 0.6 34.1 25.2 20–25

Sweet potato 68.5 1.2 0.5 25.2 24.0 10–29
Potato 63–83 0.7–4.6 0.2 36.8 32 22–24
Sugar beet 60–70 6.5 7.0 36.1 5 8–12
Yam 56.3–78.6 7.4 2.0 69.07 0.13–0.27 8–18
Aroids 55–68 1.2 0.8 42.6 0.25 12–16
6 H. Thatoi et al.
Table 4. Major tuber crops grow in the world including India and their bioethanol production.
Common Scientific Region where Annual production Region where Annual production
name name grows (worldwide) (worldwide) grows (India) (India)
Cassava M. esculenta Thailand, Africa, 250 million Kerala, Tamil nadu, 9943 thousand
Nigeria, Asia, tonnes (Incomm Andhra Pradesh, tonnes (Biofuels
Vietnam, China, ommodity Meghalaya, Assam, ethanol, 2012)
Brazil, Paraguay, Profile, Karnataka, Nagaland,
Colombia, etc. Cassava, 2011) Pondicherry, Sikkim,
Mizoram, Rajasthan,
etc. National
Horticulture Board
(2012–2013)
Sweet potato I. batata Asia, China, 106.56 tonnes Odisha, West Bengal, 1059.00 thousand
Indonesia, Africa, FAOSTAT, Uttar Pradesh, tonnes (Biofuels
Brazil, Philippines, Fevrier (2012) Chhattisgarh, Assam, ethanol, 2012)
Uganda, Nigeria, Karnataka, Madhya
Tanzania, Cuba, Pradesh, Tamil

Cuba, Argentina, nadu, Meghalaya,
etc. Bihar, etc. National
Horticulture Board
(2012–2013)
Yam D. rotundata Nigeria, Ghana, 50.0 million Odisha, West Bengal, 1.1 million tonnes
Brazil, Japan, metric tonnes Uttar Pradesh, (Biofuels
Cuba, Africa, Asia, UNFAO (2011) Chhattisgarh, Kerala, ethanol, 2012)
Jamaika, etc. Tamil nadu, Andhra
Pradesh, Meghalaya,
Assam, Karnataka,
etc.
Potato S. tuberosum Africa, Asia, 324.1 million Uttar Pradesh, West 39,858.43
Thailand, Africa, tonnes Bengal, Bihar, thousand tonnes
Nigeria, Asia, FAOSTAT, Gujarat, Punjab, (China peoples
Vietnam, China, (2011) Madhya Pradesh, Republic of
Brazil, Paraguay, Assam, Chhattisgarh, Grain and Feed
Colombia, etc. Jharkhand, Haryana, Annual, 2012)
etc. National
Horticulture Board
(2012–2013)
Arrow root Maranta arun- Southeast Asia, 77.7 million Odisha, West Bengal, 3.999 thousand
dinaceae Africa, Australia, tonnes FAO, Uttar Pradesh, tonnes (Biofuels
Marshall Islands, (2011) Chhattisgarh, Kerala, ethanol, 2012)
Nigeria, Asia, Tamil nadu, Andhra
Vietnam, China, Pradesh, Meghalaya,
Brazil Assam, Karnataka,
etc.
2.1. Cassava
Cassava (Manihot esculenta) is a tropical and subtropical perennial or temperate annual shrub
belonging to the family Euphorbiaceae, which originated from South America. An advantage of
cassava is that it can tolerate semi-arid conditions with rainfall as low as 500 mm thus have a high
productivity (O’Hair 1990). It has another advantage of growing in a wide range of soils but the
most adaptable soil is the one which is well drained and light textured. It is usually grown in soils
that are not most suitable for it, thus the fertile soils are left for other crops that are less tolerant
to poor soils. World production of cassava is estimated at over 12.5 tonnes ha−1 in the year 2011.
According to the United Nations Food and Agriculture Organization, cassava ranks fourth as a
food crop in the developing countries, after rice, maize and wheat. Africa and Asia are the leading
producers, whereas Nigeria is the largest producer of cassava (Ibeto, Ofoefule, and Agbo 2011).
The most productive cassava farms in the world are found in India, with a nationwide average
yield of 34.8 tonnes ha−1 in 2010 (Nyerhovwo 2004). A typical composition of the cassava root is
moisture (70%), starch (24%), fibre (2%), protein (1%) and other substances including minerals
(3%). The nutritive reserve of cassava is made up of starch, which is one of the most important
products that can be industrially utilised for bioethanol production (Swaider and Ware 2002).
2.2. Sweet potato
Sweet potato (Ipomoea batata) is a perennial crop in tropical and sub-tropical climates and
an annual crop in temperate regions (Kays and Wang 2002). It is a member of the Con-
volvulaceae family and native to Central and South Americas. Globally, sweet potato is the
seventh most food crop after wheat, rice, maize, potato, barley and cassava and the second most
important root and tuber crop after potato (Scott and Maldonado 1997). The global annual pro-
duction of sweet potato per year is more than 140 million tonnes. Asia is the world’s largest
potato-producing continent, with an annual production of 129 million tonnes. The total annual
production of sweet potato in India was 1046 tonnes in the year 2009–2010 (Indian Horticul-
ture Database 2011). It is a native crop of the state of Odisha (India) and available in plenty
with a total production of 438.80 tonnes (Jata et al. 2011). The proximate composition of sweet
potato is moisture (70.54%), protein (12.21%), carbohydrate (25.4%) and other components,
such as ash, fat and crude fibre in minor quantities (Abubakaret et al. 2000). Cultivars vary in
colour from white to orange and even purple depending on the starch content. Generally, white-
fleshed types are higher in starch (25–40%), less sweet, larger in size, and are not considered
a main food-source crop (Aguilera 2008). Most white-fleshed industrial sweet potatoes are edi-
ble but are not palatable due to their high starch content. Thus, these sweet potato varieties are
considered as important for bioethanol production. According to researchers at USDA’s Agri-
cultural Research Service, the potential to use sweet potatoes as a feed stock for producing
ethanol is that it produces two to three times more carbohydrates than corn (Ibeto, Ofoefule,
and Agbo 2011). Industrial sweet potatoes are bred to increase its starch content, significantly
reducing its attractiveness as a food crop when compared with other conventional food cultivars
(visual aspect, colour and taste). Therefore, these tuber crops offer potentially greater fermentable
sugar yields for industrial conversion processes. It has been reported that some industrial sweet
potatoes breeding lines developed could produce ethanol yields of 4500–6500 L/ha compared to
2800–3800 L/ha for corn (Ziska et al. 2009).
2.3. Potato
Potato (Solanum tuberosum) a member of the Solanaceae family, is a seasonal crop grown in
temperate zones all over the world, but primarily in the northern hemisphere. It is one of the
world’s most important food crops following rice, wheat and maize (Öhgren et al. 2007). The
world production of potato in the year 2011–2012 was recorded to be 330 million tonnes. China
is now the world’s one-third potato producing country with a production of 20% followed by
Russia (12%), India (8%) and USA (8%). In the year 2012–2013, the annual production of potato
in the top 10 states of India was recorded to be 14,000 tonnes (Lee et al. 2012). The total mass
of potato consists of 23.7% of dry matter, 17.5% of starch, 2% crude protein, 0.5% total sugar,
0.3% reducing sugar and other materials in traces. Potatoes are used to brew alcoholic beverages,
i.e. vodka, potcheen or akvarit. The starch from potato is used in the food industry and for the
production of bioethanol.
2.4. Yam
Yams (Dioscorea rotundata) are large starchy tubers produced annually and perennially in Africa,
America, Caribbean, South Africa and Asia and belongs to Dioscoreaceae family. According to the
8 H. Thatoi et al.
FAO 2008, the world production of yam was 50 million tons, with India accounting for 1.1 million
tons during 2008. The total moisture content(wet weight basis), starch, protein, carbohydrate, fat
and ash (dry weight basis) of yam are 10.6%, 20% 19.5%, 62.2%, 2.5% and 2.8%, respectively
(Edem, Amugo, and Eka 1990).The starch present in yam can be hydrolysed, fermented and
then distilled to produce ethanol (Wagner 2005). The principal edible yams are widely grown
throughout the tropics are white yam (D.rotundata), yellow yam (Dioscorea cayenensis), bitter
yam (Dioscorea dumetorum) and water yam (Dioscorea alata). In West Africa, they are consumed
mainly as ‘fufu’, stiff glutinous doughe (Kymalainen 2007).
2.5. Sugar beet
Sugar beets (Beta vulgaris) are a root crop which grows underground and is a biennial plant belongs
to the Amaranthaceae family. It is mostly composed of carbohydrates (66.3%) with minor amounts
of ash, proteins and lignin (Chandel et al. 2007). The world harvested 271.6 million metric tonnes
of sugar beets in 2011. The world’s largest producer is Russia, with a 47.6 million metric tonne.
The average yield of sugar beet crops worldwide was 58.2 tonnes per hectare. In 2009, France,
the USA, Germany, Russia and Turkey were the world’s five largest sugar beet producers. When
fully grown, a sugar beet weighs two to five pounds and produces about three teaspoons of sugar.
Theoretically a ton of sugar beet is said to yield 110 L ethanol, whereas a hectare area of sugar beet
gives 4200 L ethanol. When crystallised sugar obtained from beet sugar is added to corn slurry,
it is seen to enhance the fermentation process. The advantages of sugar beet are a lower cycle
of crop production, higher yield and high tolerance of a wide range of climatic variations, low
water and fertiliser requirement, which are beneficial as a low investment substrate for bioethanol
production.
2.6. Aroids
Aroid is the common name for of the Araceae family of plants, sometimes known as the Philo-
dendron or Arum family. Most aroids are tropical and include members from terrestrial, aquatic
and epiphytic habitats. Aroids are cultivated for food and industrial uses in most of the tropical
and subtropical parts of the world. Among various edible aroids, commercially cultivated promis-
ing tuber crops are Colocasia, Xanthosoma, Alocasia and Amorphophallus. Colocasia esculenta
(commonly known as taro) and Xanthosoma sagittifolium (tania) are the most important of the
edible aroids. The total moisture, protein, carbohydrate and crude fibre of taro are (63–85%),
(1.2–3%), (13–29%), (6.60–1.18%) and for tania are (70–77%), (1.3–3.7%), (17–26%) and (0.6–
1.9%), respectively. These are mostly herbaceous plants often within large root stocks that act as
storage organs (Lin and Tanaka 2006).
2.7. Jerusalem artichoke
Jerusalem artichoke (Helianthus tuberosus) is a tuberous-rooted perennial crop belongs to the

Asteraceae family and native to North America. Its tuber is rich in synanthrin and other fructose
polymers constituting of 3–60 fructose units and one glucose unit. The inulin content in fresh
tuber is about 10–20% with an average of 15%. The sugar produced in Jerusalem artichoke is
stored in the roots and tuber and is seen to have a high alcohol production potential. The advantage
of this tuber is that it does not require fertile soil for its growth and if the small tubers are left in
the field they will produce the next season’s crop, so no ploughing or seeding is necessary (Lixin
et al. 2007).
2.8. Asphodelus aestivus
Asphodelus aestivus belongs to the family of Asphodelaceae is encountered in Cyprus and other
Mediterranean countries, growing in wild pastures and its tubers contain about 10.1% of starch
(Dien, Cotta, and Jeffries 2003). It is a Geophyte (Cryptophyte) dominating in eastern Mediter-
ranean region in areas degraded due to overgrazing (Dien, Cotta, and Jeffries 2003) and are well
adapted to the peculiarities of the Mediterranean climate, thus capable of facing climatic stresses.
The bioethanol is produced by fermentation of the mash produced by crushing the tubers of the
plant. The tubers contain starch, lipids and soluble sugars (sucrose, glucose and fructose), the
content of which varies considerably during the year depending on the life phase of the plant
(Dien, Cotta, and Jeffries 2003).
3. Bioethanol production technology
Industrial production of bioethanol from tuber crops depends upon several factors, namely
feedstocks, micro-organisms and their culture types, fermentation technology and process devel-
opments for pre-treatment, hydrolysis along with fermentation and distillation, the details of which
are discussed separately along with the biotechnological tools involved in bioethanol production.
3.1. Different feedstocks for fermentation
Bioethanol can be produced from numerous sugary sources. Depending on the substrate composi-
tion and utilisation, they can be broadly classified into three major groups; (a) sucrose-containing
feedstocks, (b) starchy feedstocks and (c) lignocellulosic feedstocks. The type of feedstock chosen
for ethanol production has a significant impact on the design of fermentation process. Ethanol is
produced from a variety of sugar- or starch-containing crops, with modifications in the design of
the feedstock preparation processes. The modifications are required to accommodate the physical
properties of the feedstock as well as the nature of the carbohydrate (i.e. sugar versus starch).
3.1.1. Sucrose-containing feedstock
Sucrose-containing feedstocks are the most utilised substrate for bioethanol production (Badger
2002). These feedstocks are mostly preferred because the conversion of sucrose into ethanol is
much easier compared with starchy and lignocellulosic biomass (Martín et al. 2006). In this case,
no conversion of the feedstock is required, as the disaccharides can be directly broken down by the
micro-organisms during bioethanol production (Narendranath, Thomas, and Ingledew 2000).The
main feedstocks rich in sugar for bioethanol production include sugarcane, molasses, sugar beet,
sweet sorghum and various fruits. In spite of this simple procedure, the processing of these
substrates is generally too expensive for bioethanol production, because of the availability and
transport costs of its feedstocks (Badger 2002).
3.1.2. Starchy materials
Starch is a high-yielding feedstock for bioethanol production (Apar and Ozbek 2004). Starch
molecule is a polysaccharide, made up of long chains of glucose molecules, for which it is
necessary to break down the long chains of this carbohydrate to the simpler form i.e. glucose.
Glucose can further be converted into ethanol by micro-organisms. Examples of starchy materials
10 H. Thatoi et al.
commonly used for bioethanol production in worldwide are cereal grains, corn, potato, sweet
potato, cassava, etc.
3.1.3. Lignocellulose biomass
Among the other different feedstocks, lignocellulosic feedstock is an attractive option for
bioethanol production (Sun and Cheng 2002). As the lignocellulosic complex is made up of
a matrix of cellulose and lignin bound by hemicelluloses, the main challenge in this case is to
reduce the degree of crystallinity of the cellulose and increase the fraction of amorphous cellu-
lose by the process of pre-treatment, the most suitable form for the hydrolysis step (Palmqvist
and Hahn-Hägerdal 2000). After pre-treatment, the cellulose undergoes enzymatic hydrolysis in
order to obtain glucose that is converted to ethanol by micro-organisms (Lynd et al. 2002). In
general the lignocellulosic materials can be broadly divided into six main groups for fuel ethanol
production i.e. crop residues (cane bagasse, corn, wheat straw), hard wood (aspen, poplar), soft
wood (pine, spruce), cellulose wastes (newsprint, recycled paper sludge), herbaceous biomass
(alfa alfa, switch grass) and municipal solid wastes (MSW) (Sánchez and Cardona 2008).
3.2. Micro-organisms in bioethanol fermentation
Ethanol fermentation is a biological process in which sugar (hexose) is converted to ethanol by

micro-organisms. During the whole process of ethanol fermentation, micro-organisms participate
in two major applications. In one application, micro-organisms convert fermentable substrates into
ethanol and in the other micro-organisms produce the enzymes to catalyse conversion of com-
plex carbohydrates into simpler sugars (Skoog and Hahn-Hägerdal 1988). Several reports and
reviews have been published on production of ethanol through fermentation by micro-organisms
where several bacteria, yeasts and fungi have been reportedly used (Dasgupta et al. 2013). These
microbes are capable of yielding ethanol as the major product. Historically, the most commonly
used microbe in bioethanol production has been yeast. Among the yeasts, Saccharomyces cere-
visiae (S. cerevisiae) is the preferred one, as it can produce high concentration (18%) of ethanol in
the fermentation broth (Zhao and Bai 2009). They have been the centre of attraction for bioethanol
research since the dawn of biorefinery processes. They can grow on variety of substrates with
high sugar content and have high ethanol tolerance capacity. Although they grow preferably on
hexose sugars, few strains are also capable of utilising pentose sugars converting into ethanol
(Nigam 2001). Among the pentose utilising yeasts, species such as Pichia tannophilus, Can-
dida shehatae, P. stipites and Kluveromyces maxianus are well known for ethanol fermentation.
Generally it was observed that ethanol production by P. tannophilus and species belonging to
Candida and Pichia were better on rich media under aerobic conditions. It has been noticed
that majority of yeasts do not ferment D-xylose directly, instead they utilise D-xylulose, an
isomer of D-xylose, both oxidatively and fermentatively (Rosenberg 1980). The best xylulose-
fermenting yeasts so far identified are species of Brettanomyces sp., Candida, Hansenula and
Torulospora. Pentose utilising yeasts also convert xylose into xylitol in addition to ethanol. How-
ever, an important yeast, P. stipites apparently produces no xylitol during sugar fermentation,
which is a unique quality of this micro-organism (Deshpande et al. 1986). The ability of fil-
amentous fungi to ferment pentose sugars has been known for about 70 years. Several fungal
species belonging to genera Fusarium, Rhizopus (Hahn-Hägerdal et al. 2007), Monilia (Singh,
Kumar, and Schugerl 1992), Neurospora (Xiros and Christakopoulos 2009) and Paecilomyces
were found to have potential for fermenting glucose as well as xylose. In most fungi, the initial
conversion of D-xylose and L-arabinose to D-xylulose 5-phosphate proceeds through a series of
reduction and oxidation steps involving the cofactors NAD (P)+/NAD (P) H (Sommer, Georgieva,
and Ahring 2004). Among the ethanol producing fungi, Fusarium oxysporum is reported to have
shown more ethanol production than Neurospora crassa and Mucor sp (Pearlman 1950). Few fun-
gal strains have been identified that ferment not only glucose and xylose but also other complex
natural cellulosic substrates, which is an advantage to get the maximum yield of ethanol (Detroy
and Bolen 1983).
Unlike yeast and filamentous fungi which do not ferment sugars anaerobically, bacteria can
readily convert xylose to ethanol under anaerobic conditions (Sonneitner 1983). These include
Zymomonas mobilis, Bacillus macerans, B. polymyxa, Kiebsiella pneumoniae, Clostridium aceto-
butylicum, Aeromonas hydrophila, Aerobacter sp., Erwinia sp., Leuconostoc sp. and Lactobacillus
sp. (Millichip and Doelle 1989). These micro-organisms convert sugars into ethanol along with
undesired by-products such as acetic acid, lactic acid, 2, 3-butane-diol and CO2 . In addition
to mesophilic bacteria, thermotolerant bacterial species, such as Clostridium thermocellum, C.
thermohydrosulfurium, C. thermosaccharolyticum, C. thermosulfurogenes and Thermoanaero
bacteretahnolicus (Senthilkumar and Gunasekaran 2005), can also produce ethanol. Z. mobilises
an unusual Gram-negative bacterium has several appealing properties as a biocatalyst for ethanol
production. The micro-organism has an ethanol fermentation pathway and tolerates up to 120 g/L
ethanol. The high ethanol yield and productivity observed for Zymomonas are a consequence of
its unique physiology (Ado et al. 2009). Engineering Escherichia coli is another valuable bacterial
resource for ethanol production. The construction of E. coli strains to selectively produce ethanol
(Tran and Chambers 1986) was one of the first successful applications of metabolic engineer-
ing. Though bacteria grow faster in relation with fungi and yeast, their low substrate specificity
do not make them the right choice for ethanol production at large scale from sugar substrates
(Srichuwong et al. 2009).
3.3. Culture types used in fermentation
In fermentation, microbial culture types play an important role for efficient ethanol production.
Ethanol fermentation from tuber crops as substrates is carried out by different culture types. There
are mainly five different culture types that can be utilised for fermentation. These include pure
culture, co-culture, mixed culture, immobilised culture and co-immobilised culture. The culture
types with their examples are given in Table 5.
Pure culture of an organism is the culture that contains large population of only one type
of organism generally developed from a single cell. Processes with pure cultures are normally
targeted on a single product. Pure cultures rarely exist in nature as the micro-organisms are present
in mixed cultures in nature. Therefore, pure cultures are artificially obtained in in vitro condition.
Pure culture can be advantageous in processwise if two or more products are produced by the
culture technique, the utilisation of substrate will be more and the costs of product recovery could
be significantly reduced (Han et al. 2011). A study on bioethanol fermentation from cassava stalks
was carried out by Sovorawet and Kongkiattikajorn (2012) using the pure culture of S. cerevisiae
and the ethanol yields were 98.43% and 95.29% in simultaneous hydrolysis and fermentation and
simultaneous saccharification and fermentation (SSF), respectively. In another study, bioethanol
production from raw sweet potato by pure culture of S. cerevisiae at laboratory, pilot and industrial
scales was studied by Zhang et al. (2011). The maximum ethanol concentration, average ethanol
productivity and ethanol yield after fermentation in laboratory scale were 128.51 g/L, 4.76 g/L/h
and 91.4% with a small decrease at pilot scale (109.06 g/L, 4.89 g/L/h and 91.24%) and industrial
scale (97.94 g/L, 4.19 g/L/h and 91.27%), respectively. Though the ethanol productivity is high,
the whole substrate utilisation remains a major concern in pure culture techniques. This is a major
issue which is thought to be eliminated by co-culture of micro-organisms.
Co-culture is a culture that contains growths of two distinct cell types. It is useful in under-
standing the interaction of two different cell types. Ethanol yield by co-culture has been better
12 H. Thatoi et al.
Table 5. Microbial culture types used in fermentation for bioethanol production from tuber crops.
Organisms Substrate Treatment Reference
Pure culture
Saccharomyces cerevisiae Potato Enzymatic hydrolysis Khan et al. (2012); Akponah
and Akpomie (2012)
Acid hydrolysis Akponah and Akpomie (2012)
Potato starch Steam Treatment
Potato mash Enzymatic hydrolysis Srichuwong et al. (2009)
Potato flour Enzymatic hydrolysis Rani et al. (2010)
Sugar beet root Alkali treatment Srichuwong et al. (2009)
Enzymatic hydrolysis Padmaja, George, and
Moorthy (2006)
Sweet potato root – Ziska et al. (2009)
Cassava tuber – Ziska et al. (2009); Padmaja,
George, and Moorthy
(2006)
Cassava root peels Enzymatic hydrolysis Akponah and Akpomie (2012)

Acid hydrolysis
Yam Enzymatic hydrolysis
Acid hydrolysis
Engineered Z. mobilis Raw sweet potato starch – Zhang et al. (2009)
Z. mobilis Raw sweet potato Enzymatic hydrolysis He et al. (2009a)
Yeast Sweet potato Enzymatic hydrolysis
Co-culture
Saccharolytic Molds and S. Sweet potato Enzymatic hydrolysis Yang et al. (2012)
cerevisiae
Saccharomyces diastaticus Cassava peels Enzymatic hydrolysis Kongkiattikajorn and
2047 and S. cerevisiae7532 Sornvoraweat (2011)
Rhizopus sp. and S. cerevisiae Raw sweet potato starch Enzymatic hydrolysis Yuwa-Amornpitak (2010)
Paenibacillus sp. and Z. Raw sweet potato starch Simultaneously versus He et al. (2009b)
mobilis subsequently co-cultured at
48 h of fermentation time
(100 mL shake flask)
Aspergillus niger and Potato starch – Jeon et al. (2007)
S. cerevisiae
S. cerevisiae and Uncooked raw starch – Chen et al. (2007)
Streptococcus bovis
Endomycopsis fibuligera Cassava starch Reddy and Basappa (1996)
NRRL 76 and Z. mobilis
ZM4
Saccharomycopsis fibuligera Potato starch Enzymatic hydrolysis Abouzied and Reddy (1987)
and S. cerevisiae
Aspergillus niger S. Potato starch Enzymatic hydrolysis Abouzied and Reddy (1986)
cerevisiae
Clostridium thermohydro 5% Starch with TYE 14 L microfermentor Hyun and Zeikus (1985)
sulfuricum Clostridium medium (contains
thermo sulfurogenes vitamin solution,
ammonium chloride,
magnesium chloride and
trace mineral)
Mixed culture
Paenibacillus sp. and four Sweet potato – Zhang et al. (2011)
strains of Z. mobilis
S. cerevisiae and Aspergillus Potato starch Enzymatic saccharification Jeon et al. (2007)
niger
10 species of Rhizopus sp. Cassava starch –
and S. cerevisiae
S. cerevisiae CFTRI 101, Casava starch Enzymatic hydrolysis Reddy and Basappa (1996)
S.diastaticus IFO 1046,
Endomycopsis fibuligera,
Schwanniomyces castellii
ATCC 26077 and Z.
mobilisZM4
(Continued)
Table 5. Continued.
Organisms Substrate Treatment Reference
Immobilised cultures
Saccharolytic molds Sweet potato Enzymatic treatment Lee et al. (2012)
(Aspergillus oryzae and
Monascus purpureus)
with S. cerevisiae
Bacillus brevis MTCC Cassava fibrous residue Enzymatic treatment Ray and Moorthy (2007)
7521, Lactobacillus
plantarum MTCC 1407
and S. cerevisiae RC
CTCRI
Mixed culture of S. Jerusalem artichoke Acid and thermal treatments Williams and Ziobro (1982)
cerevisiae and Z. mobilis
compared to pure culture (Nuwamanya et al. 2012). The feasibility of ethanol production from
sweet potato root flour (SPRF) was studied by Swain, Mishra, and Thatoi (2013) using a co-culture
of Trichoderma sp. and S. cerevisiae. In this experiment, the maximum ethanol production by the
co-culture of these two organisms was found to be 172 g/kg substrate with 10% inoculum size
and fermented at 30◦ C for 72 h. Cell interaction was observed advantageous in case of co-culture
and the ethanol production ability by the co-culture was observed to be 65% higher than the single
culture of S. cerevisiae from un-saccharified SPRF. In another study by Jirasak and Buddhiporn
(2011), bioethanol production from cassava peels pre-treated with diluted sulphuric acid was
studied using mono cultures of Saccharomyces diastaticus and S. cerevisiae and a co-culture of
S. diastaticus and Candida tropicalis by SSF. The study revealed that co-culture of S. diastaticus
and C. tropicalis produced greater amounts of ethanol than those of mono cultures.
There is another type of culture known as mixed culture, which utilises more than two organisms.
Mixed culture usually holds two or more identified and easily differentiated species of micro-
organisms (Vijaya, Sarathi, and Basappa 1996). In mixed culture many organisms may be used
in which each organism could use a specific substrate and produce a specific product in the same
bioreactor. Mixed cultures can also have several other advantages over pure cultures and co-
cultures (Delenges, Moletta, and Navarro 1996). A primary application is that a mixed culture of
micro-organisms given to different enzymatic systems and combination of metabolic pathways
increases the efficient utilisation of substrates and also the product yield. For example, a typical
problem of many fermentation processes with pure cultures is the production of by-products in
form of organic acids or alcohols, which are toxic to cell growth. In a bioprocess with a mixed
culture, the toxic by-products could be degraded or even converted to another useful product by
one of the species, leading to bioprocessing with multiple products and a more efficient use of the
substrate(s). This is exactly the goal of the new concept biorefinery that has been proposed as a
future direction of industrial biotechnology. Mark and Loosdrecht (2007) investigated that mixed
culture biotechnology (MCB) could become an attractive addition or alternative to traditional pure
culture-based biotechnology for the production of bioenergy. On the basis of ecological selection
principles, MCB-based processes can generate a narrow product spectrum from a mixed substrate.
Szambelan, Nowak, and Czarnecki (2004) used Kluyveromyces fragilis, a yeast with either a
commercial yeast, S. cerevisiae or the bacterium Z. mobilis producing an ethanol concentration of
0.48 g/g for the mixed population and 0.46 g g/1 for the single population. The theoretical yield
of the mixed cultures was observed to be 2–12% higher than that of the single micro-organism.
Micro-organisms cannot only be used as free cells but can also be utilised in entrapped manner.
This technique of cell localisation which allows the entrapment of the micro-organisms within a
given matrix is known as immobilisation. Immobilisation by adhesion to a surface (electrostatic
14 H. Thatoi et al.
or covalent), entrapment in polymeric matrices or retention by membranes has been successful

for ethanol production (Szambelan, Nowak, and Czarnecki 2004). Immobilised cells offer rapid
fermentation rates with high productivity, without risk of cells being washed out. The loss of
intracellular enzyme activity can be kept to a minimum level by avoiding the removal of cells from
downstream products. The applications of immobilised cells have made a significant advancement
in fuel ethanol production technology (Gksungur and Gven 1999). Immobilisation of microbial
cells for fermentation has been developed to eliminate inhibition caused by high concentration of
substrate and also enhance ethanol productivity and yield (Wen et al. 2002). The immobilisation
of organisms can be done on sodium alginate, calcium alginate or poly acrylamide gels, but
immobilised beads made up of polyacrylamide gels have an advantage over other bead-making
substances. One such example was shown by Calinescu et al. (2012) who studied production
of bioethanol from beet molasses, by immobilised S. cerevisiae yeast. The immobilisation of
yeast on polyacrylamide gel proceeds with an efficiency comparable to the one obtained for the
immobilisation of the yeast cells on calcium alginate. The advantage of using immobilised yeast
on polyacrylamide gel is that the granules are more resistant and they keep their shape during the
fermentation process. There are many reports of efficient ethanol production from immobilised
culture and co-immobilised culture. One such report was given by Lee et al. (2012) in which
bioethanol production from sweet potato by both immobilised cells and co-immobilised cells of
S. cerevisiae and saccharolytic moulds (Aspergillus oryzae and Monascus purpureus) was studied.
In co-immobilisation of S. cerevisiae with A. oryzae and M. purpureus, the co-immobilised cells
produced 46.7 g/L ethanol from 150 g/L liquefied cassava starch, while immobilised cells of yeast
S. diastaticus produced 37.5 g/L ethanol. In repeated-batch fermentation using co-immobilised
cells, the ethanol concentration increased to 53.5 g/L. The co-immobilised gel beads were stable
up to seven successive batches. Continuous fermentation using co-immobilised cells in a packed
bed column reactor operated at a flow rate of 15 mL/h (residence time, 4 h) exhibited a maximum
ethanol productivity of 8.9 g/L/h. The immobilisation of micro-organisms definitely proves to
be an efficient culture type for tuber crops, which in future can be utilised for obtaining a more
cost effective process for the production of bioethanol.
3.4. Fermentation techniques
Fermentation is one of the oldest processing technologies in the world. Although in the past
fermentation of food and beverages was carried out naturally, without a fermenter. As a
result, fermentation process was not efficient, as appropriate parameters were not maintained.
Today with the technological advancements have made the fermentation process more effi-
cient and valuable. Like any other fermentation processes, the fermentation of tuber crops for
bioethanol production can be achieved by three processes, namely batch, fed batch and continuous
fermentation.
Batch culture is a fermentation technique in which growth takes place in a closed-loop sys-
tem. Fermenter is filled with the prepared mash of raw materials to be fermented (Zhao and Bai
2009). The temperature and pH for microbial fermentation is properly adjusted and occasionally
nutritive supplements if any are added to the prepared mash. The mash is steam-sterilised and
the inoculum is added to the fermenter, from a separate culture vessel. Fermentation proceeds
and after the proper time the contents of the fermenter are taken out for processing. The fer-
menter is cleaned and the process is repeated. Thus, each fermentation cycle is a discontinuous
process divided into batches. Batch culture can be divided into four distinct phases; lag phase,
exponential or logarithmic phase, stationary phase and death phase (Olsson and Hahn-Hägerdal
1996). When micro-organisms are introduced into fresh culture medium, usually no immediate
increase in cell number occurs. This period is called the lag phase, which is followed by the
exponential (log) phase and stationary phase. The microbial rate of growth is constant during the
exponential phase and in a closed system such as a batch culture, population growth eventually
ceases to a stationery phase. Finally, the micro-organisms enter into the death phase where no
more products are formed (Shama 1988). The batch systems consist of tanks that are designed in
relation to fermentation tank capacity and holding time. The tank is equipped with heat exchang-
ers, agitators, mixing impellers and a motor. Batch culture is useful where the shelf life of the
end product is short and is specifically used for the product produced only at the stationary phase.
Bioethanol production from industrial sweet potatoes was investigated by William et al. (2013),
where they examined liquefaction, saccharification and fermentation of industrial sweet potatoes
(ISPs) using α-amylase and glucoamylase for the production of ethanol by batch fermentation
technique. Yeast fermentation on hydrolysed starch was examined over time with and without the
addition of salt nutrients which converted all fermentable sugar (e.g. glucose, fructose, maltose)
and produced 62.6 and 33.6 ethanol g/L of hydrolysate for flour (25% w/v, substrate loading)
and fresh (12.5% w/v, substrate loading) industrial sweet potatoes respectively, after 48 h with-
out any salt addition. Grahovac et al. (2012) studied bioethanol production in batch culture by
free S. cerevisiae cells from intermediates of sugar beet processing. The ethanol yield from the
media based on intermediates of sugar beet amounted to 490 g/kg. The feasibility of bioethanol
production from yam, potato and cassava root peels by batch fermentation was investigated by
Akponah and Akpomie (2012). Liquefaction and saccharification was followed by subsequent
fermentation by S. cerevisiae at room temperature for 72 h. The highest ethanol production was
from batch fermentation of cassava root peels, followed by potato peels while yam peels yielded
the least ethanol concentration. The batch fermentation of potato tubers flour was carried out
by Rani et al. (2010) with a production of 56.8 g/L ethanol. Batch systems can be modified to
fed-batch cultures to give better productivities and to lower manufacturing costs.
Fed-batch fermentation is a production technique in between batch and continuous fermen-
tation. A proper feed rate, with the right component constitution is required during the process
(MarèlneCot, Loret, and Francois 2007). Fed-batch process is a more efficient cultivation strategy
than the batch process in which micro-organisms work at low substrate concentration with an
increasing ethanol concentration. Fed-batch cultures often provide better yield and productivities
than batch cultures by preventing contaminations (Wang et al. 2007). A strategy for preventing
reduction in ethanol yield in fermentation caused by bacterial contamination was developed by
(Kumar et al. 2009). In this fed-batch fermentation in which saccharification, fermentation and
ethanol recovery were performed simultaneously, the addition of exogenous ethanol to the fer-
mentation mixture at the start of fermentation at 50 g kg−1 prevented contamination, and the
ethanol yield reached 0.50 g g−1 . Kim et al. (2013) studied bioethanol production from Jerusalum
artichoke using Kluyveromyces marxianus in batch and fed-batch fermentation by SSF of both
pre-treated stalk and tuber yield 29.1 and 70.2 g/L ethanol, respectively. It was observed that in
fed-batch fermentation, ethanol productivity was 0.255 g ethanol per gram of dry Jerusalem arti-
choke biomass. A simple process for on-farm bioethanol production from cassava, using cassava
koji supplemented with crude liquid enzyme and yeast was described. A fed-batch mode was
done for a mixture of koji and gelatinised cassava flour to which 30 g of yeast cells were mixed
and allowed to ferment for two days, followed by the addition of cassava flour and fermenting
for another three days. The fed-batch mode gave a higher ethanol concentration of 7.05% (0.34
ethanol g/g cassava flour) (Ogbonna and Okoli 2010).
Another efficient fermentation technique is the continuous fermentation. In continuous fer-
mentation, the substrate is added to the fermenter continuously at a fixed rate. This maintains
the micro-organisms in the logarithmic growth phase. The fermentation products are taken out
continuously. The advantages of continuous culture technique over batch and fed-batch systems is
that the percentage of end product in continuous culture is much higher than batch and fed-batch
systems and the continuous process eliminates much of the unproductive time associated with
16 H. Thatoi et al.
cleaning, recharging, adjustment of media and sterilisation. A high cell density of microbes in
the continuous fermenter is locked in the exponential phase, which allows high productivity and
overall short processing of 4–6 h as compared to the conventional batch fermentation (24–60 h).
This results in substantial savings in labour and minimises investment costs by achieving a given
production level with a much smaller plant. But the only disadvantage is that the complete ster-
ilisation is very difficult in this process (Alexender, Chapman, and Jeffries 1989). Continuous
fermentation using glucose as a substrate was studied by Godoy et al. (2008) using 12 differ-
ent Saccharomyces sp. yeast strains with flocculent. The system reached ethanol yield levels of
83.53% theoretically with a maximum total reducing sugars conversion of 92.68%. These fer-
mentation techniques have a great impact on the bioethanol yield from the tuber crops and the
type of technique used can be altered depending on the type of substrate and micro-organisms
used.
3.5. Process development
Process development is an amalgamation of different operations for a particular process that

are generally considered in terms of process design and optimisation. This is valid for all the
fermentation processes irrespective of the feedstock used (Balat and Balat 2009), but the process
is specific for each separate feedstock, i.e. a specific process is used for lignocellullosic biomass
and similarly another specific process is adopted for starchy materials such as tuber crops. The
general process design ideally used for tuber crops includes five aspects, which can be integrated
to develop a process that is economical and productive, as shown in Figure 3. They are: pre-
treatment and hydrolysis of the substrate, the fermentation process, optimisation of parameters,
and finally a simulation study which can integrate all these aspects and define a process that can
be practically used to get a valuable product.
Raw material
receiving and washing
Crushing
Drying and milling
Dextrinization
Liquefication
Saccharification
Fermentation
Ethanol recovery
Product storage
Figure 3. Block diagram for the fuel ethanol production from tuber crops.
3.5.1. Pre-treatment
Pre-treatment basically refers to the mechanical and physical actions to clean and shape the
biomass and destroy its cell structure to make it more accessible with for further chemi-
cal or biological treatment (Alvira et al. 2010). The pre-treatment step changes the native
properties of the substrate in order to prepare the materials for enzymatic degradation.
Tuber crops generally consist of starch, which are molecules made up of long chain of
glucose molecule (Yang and Wyman 2008). Thus starchy substrates can be fermented after
breaking starch molecule in to simpler glucose molecules and this can be done by a pre-
treatment step. The pre-treatment step changes the native properties of the substrate in order
to prepare the materials for enzymatic degradation (Wyman et al. 2005). Pre-treatment meth-
ods can be categorised into physical, chemical, biological and physicochemical (Chandel
et al. 2010). In physical pre-treatment, mechanical milling of the desired substrate is carried out
for grounding the substrate. The main chemical pre-treatment methods include ozonolysis, acid
hydrolysis, alkaline hydrolysis (Wang et al. 2009) and organosoly based processes (Zhao, Cheng,
and Liu 2009). The biological pre-treatment methods mainly involve utilising different fungal
species (Farrell et al. 2006) and the physicochemical pre-treatment methods include ammonia
fibre explosion (Prasad, Singh, and Joshia 2007) and steam pre-treatment (Srichuwong et al.
2009). In case of starchy substrate, a mechanical milling is required, where the requirement is of
the flour of the whole tubers. The mechanical milling grounds the dried tubers to a certain particle
size that can pass through a mesh screen and then the flour can be collected and used for further
processing i.e. hydrolysis of the substrate. Generally tuber crops are subjected for enzymatic, acid
and biological hydrolysis. When starch is heated in water, it forms a very thick gel. This is called
as gelatinisation. Starch gelatinisation is essential for efficient enzymatic hydrolysis (Takashi et al.
2010). The gelatinisation is followed by hydrolysis of starch.
3.5.2. Enzymatic hydrolysis
Hydrolysis is a chemical reaction in which a compound will react with water to produce other
compounds. Enzymatic hydrolysis is one of the basic methods for hydrolysis. Enzymes are nat-
urally occurring plant proteins that cause certain chemical reactions to occur (Castaño et al.
2011). However, for enzymes to work, they must obtain access to the molecules to be hydrol-
ysed (Badger 2002). Generally, the starchy substrates require a reaction of starch with water and
enzymes (hydrolysis) to break down the starch into fermentable sugars. Enzymes like α-amylase
and glucoamylase (Teramoto, Lee, and Endo 2008), which are responsible for the breakdown of
the chemical bonds in the starch, are given at various times during the heat treatment (Badger
2002). The dextrinisation of the starch is done by using α-amylase which randomly hydrolyses
internal α-1,4-glucosidic bonds in starch and its degradation products, liberating soluble dextrins
and oligosaccharides that are more suitable for efficient conversion to glucose. It is followed by
saccharification that is carried out by glucoamylase which hydrolyses 1,4 and 1,6-alpha linkages
in liquefied starch to glucose (Van Voorst et al. 2006).
An efficiency of enzymatic hydrolysis for production of bioethanol from cassava starch using
new enzymes like Spezyme® Xtra and Stargen™ was studied by Shanavas et al. (2011). The
liquefying enzyme Spezyme was optimally active at 90◦ C and pH 5.5 on a 10% (w/v) starch
slurry at levels of 20.0 mg (280 Amylase Activity Units) for 30 min. Stargen levels of 100 mg
(45.6 Granular Starch Hydrolysing Units) were sufficient to almost completely hydrolyse 10%
(w/v) starch at room temperature (30 ± 1◦ C). Ethanol yield and fermentation efficiency were very
high (533 g/kg and 94.0%, respectively) in the Stargen + yeast process with 10% (w/v) starch for
48 h. The specific advantage of the new process was that the reaction could be completed within
48.5 h at 30 ± 1◦ C. In another study, bioethanol production from potato wastes using mixture
18 H. Thatoi et al.
of digesting enzymes after saccharification and fermentation was studied by Khan et al. (2012).
This investigation revealed that mixture of enzymes significantly enhanced bioethanol production
compared to non-treated mixture.
3.5.3. Chemical hydrolysis
Conversion of starchy materials to glucose monomers by saccharification process can be achieved

by acid and enzyme hydrolysis. The enzymatic hydrolysis is a long-timed process and currently
the cost of enzymes is also too high and research is continuing to bring down the cost of enzymes
(Badger 2002). So, acid enzyme hydrolysis method is used. There are two basic types of acid
hydrolysis processes; dilute acid and concentrated acid, each with variations. Dilute acid hydrol-
ysis processes are conducted under high temperature and pressure and have reaction times in the
range of seconds or minutes (Badger 2002). Acids used for chemical hydrolysis include H2 SO4 ,
HCl, H2 O2 , phosphoric acid and nitric acid, etc. In the process, the acid first breaks down the
matrix structure of the fibre into more accessible cellulose, hemicellulose and lignin, and then
further transforms these polysaccharides to monosaccharides. This type of application commonly
uses either concentrated acid, 70–77% (w/w), at a low temperature, around 50◦ C, or diluted
acid, 0.4–0.7% (w/w), at a high temperature (Sun and Cheng 2002). After conversion to sim-
pler monomers (glucose), it is fermented to produce ethanol with the help of ethanol-producing
micro-organisms (Lin and Tanaka 2006). It was also observed that dilute solutions of hydrogen
peroxide can be employed for depolymerisation of the polysaccharide molecule, such as dextran,
over a wide range of temperature, pressure, time and concentration of hydrogen peroxide. Thus
it was therefore possible to use hydrogen peroxide in place of the usual hydrolytic agents, such
as acids, for the depolymerisation of native dextran to smaller molecular weight fragments.
Bioethanol production from potato starch residue (stream produced during chips manufactur-
ing) treated with 1% H2 SO4 using S. cerevisiae was studied by Hashem and Darwish (2010).
Results demonstrated that 1% H2 SO4 at 100◦ C for 1 h was enough to hydrolyse all starch con-
tained in the residue stream. The maximum yield of ethanol of 5.52 g/L was achieved at 35◦ C
by S. cerevisiae y-1646 after 36 h. Marija et al. (2009) also studied the hydrolysis of starch from
fresh potato tubers by HCl and H2 SO4 at different ratios of plant material to acid solution. The
ethanol yield of 31 g/L was obtained in the fermentation of hydrolysate prepared under the optimal
hydrolysis conditions by commercial bakery yeast at 28◦ C for about 18 h. Recently, bioethanol
production by separate hydrolysis and fermentation method from cassava tubers using sulphuric
acid was studied by Echegi, Ejikeme, and Ejikeme (2013). In this study, gelatinised cassava sam-
ple was treated with the prepared solution of sulphuric acid. The effect of acid strength on the
ethanol yield was studied for acid concentrations of 1M and 3M. On acid hydrolysis, the used
sulphuric acid acted as a chemical catalyst for the breakdown of starch into glucose which was
fermented to ethanol. It was evident from the study that the increase in the concentration of the
acid speeds up the rate of hydrolysis of flour.
3.5.4. Fermentation and distillation
The saccharified biomass that is obtained after hydrolysis is then subjected to fermentation.
Fermentation is the slow decomposition of large organic molecules (such as starch) into smaller
molecules, such as ethanol. Ethanol fermentation can be described as the biochemical process
by which sugar, such as glucose; fructose and sucrose, are converted into cellular energy there
by producing ethanol and carbon dioxide as metabolic waste products (Shanavas et al. 2011).
Fermentation typically requires 12–72 h of incubation period depending on the amount and type
of micro-organism used to start fermentation. In general, root and tuber crops are preferred for
the bioethanol production due to high starch content (16–24%) (Hashem and Darwish 2011).
The ethanol fermentation processes from starchy materials commonly involves two stages: (i)
liquefaction of starch by amylase and enzymatic saccharification of the low molecular weight
liquefaction products such as dextrin to produce glucose and (ii) fermentation of glucose to
ethanol. The liquefaction, saccharification and fermentation can also be carried out simultaneously.
The advantage of such a developmental process for simultaneous liquefaction, saccharification
and fermentation of starch is that it would reduce the energy input and increase the efficiency
of substrate utilisation (Plessas et al. 2007). When certain species of micro-organisms (most
importantly yeasts) metabolise sugar in the absence of oxygen, they produce ethanol and carbon
dioxide from tuber crops.
After fermentation, the ethanol and water from the sugar-containing liquid mixture are to be
separated into their components using distillation. Distillation is the most widely used separa-
tion operation in chemical and petrochemical industries accounting for around 25–40% of the
energy usage (Xiu and Zeng 2008). One disadvantage of distillation process is the large energy
requirement. Distillation consumes a great deal of energy for providing heat to change liquid
to vapour and condense the vapour back to liquid at the condenser. Distillation is carried out in
distillation columns, which consumes substantial amount of energy (Plessas et al. 2007). With
rising energy awareness and growing environmental concerns there is a need to reduce the energy
use in industry. The potential for energy savings exists in design and operation of energy efficient
distillation systems.
Bioethanol production from SPRF was studied by Swain, Mishra, and Thatoi (2013) by using
co-culture of fungus Trichoderma sp. and yeast S. cerevisiae. In this experiment, the maximum
ethanol production was 172 g/kg substrate with 10% inoculum size fermented at 30◦ C for 72 h.
Lee et al. (2013) reported that co-immobilisation of S. cerevisiae with Aspergillus oryzae or
Monascus purpureus produced 46.7 g/L ethanol from 150 g/L of liquefied cassava starch, after
72 h incubation, while immobilised cells of yeast S. diastaticus produced 37.5 g/L ethanol. In
repeated-batch fermentation using co-immobilised cells, the ethanol concentration increased to
53.5 g/L. Hashem and Darwish (2010) studied bioethanol production from 1% H2 SO4 -treated
potato starch residue using S. cerevisiae. The maximum yield of ethanol of 5.52 g/L was achieved
at 35◦ C by S. cerevisiae y-1646 after 36 h.
Optimisation is a technique of making a methodology or system as fully perfect, functional and
effective as much as possible (Tokgoz 2008). In a typical optimisation problem, the goal is to find
all the optimum parameters that can be utilised to obtain the maximum productivity and minimum
waste, the production process and an investment scheme. The optimisation process can be broadly
divided into two types: (a) the conventional process and (b) the computational (mathematical and
statistical approach) techniques.
Process synthesis procedures can be significantly enhanced using process simulation pack-
ages. These simulators have allowed the analysis of several technological options and gaining
insight into the process improvements. The proper assimilation of all the optimised parameters
with technologically advanced simulation technologies will not only enhance the productivity of
bioethanol from the starchy materials, but will also allow its implementation in an industrial scale.
4. Biotechnological tools for bioethanol production from tuber crops
In the twenty-first century, production of biofuels from plant biomass are being seriously viewed
from a number of perspectives that include depleting of fossil fuel resources, agrarian economy
and new avenues of gainful employment. In recent times, blending of petrol with bioethanol has
become important to meet the fuel demand. Blending of ethanol with petrol started in India from
the year 2000 and continues till today (McMillan 1997). During 2003, the Ministry of Petroleum
20 H. Thatoi et al.
and Natural Gas has made it compulsory to supply 5% ethanol-blended petrol in a few states in
India. It has also taken steps to extend this practice to the whole country and expects to increase
the percentage of ethanol mixture in petrol to 10% (David, Alice, and Ramesh 2008).
With the depletion of fossil fuel, there is a great need for increase in bio-based ethanol pro-
duction for which new technological advancements is most essential. In case of tuber crops, the
technical methods to produce biofuel on a large scale have many challenges. It is expected that
the use of biotechnology can potentially address many of these technical challenges and envi-
ronmental concerns. In recent years, there is a significant progress in the use of biotechnological
techniques such as molecular biology, in silico genome technique, protein engineering, genetic
engineering to increase the activity of enzymes and the microbes for enhanced biofuel production
(Chandel et al. 2011). This progress will certainly help us to make bioethanol a replacement for our
present transportation fuel. Several biotechnological tools such as recombinant DNA, metabolic
engineering and mutagenesis are available for the improvement of yeast strains capable of with-
standing the harsh environments typically of certain industrial fermentation process (Neves et al.
2006).
In silico genome-scale cell model is a promising tool, which is used for accelerating the
design of cells with improved and desired properties for bioethanol production in fermenta-
tion. For example, Bro et al. (2006) reconstructed a metabolic network using a genome of
S. cerevisiae, which leads to decreased glycerol and increased ethanol yields (10%). Protein
engineering is another new approach that has put forth new avenues to create genetically engi-
neered micro-organisms (GEMs) that can function as a booster biocatalyst (Pasha, Kuhad,
and Rao 2007). Protein engineering offers powerful opportunities to enhance the efficiency
of enzymatic hydrolysis with simple modifications to the amino acid sequences of a protein
(Zhong, Zhang, and Zheng 2006). These processes have dramatic impacts on the perfor-
mance of bioethanol production from tuber crops (Zhang, Zhu, and Li 2009). Zhang et al.
(1995) reported that the use of genome shuffling for obtaining yeast strains have improved ethanol
production in fermentations with high glucose contents (200–300 g/L). There are also reports that
address the identification of genes that confer resistance to ethanol in yeast (Chandel et al. 2011) as
well the molecular and physiological responses of yeast during alcoholic fermentation particularly
under industrial conditions (Van Voorst et al. 2006). With the advancement of biotechnology, it
has been possible to genetically modify S. cerevisiae so that it can ferment both C5 and C6 sugars,
which will be useful to construct improved vigorous strains for the industry (Mussatto et al. 2010).
Although bioethanol production from substrates such as tuber crops has been greatly improved by
newly developed GEMs, there are still many more challenges (Takahashi, Shimoi, and Ito 2001).
It is expected that in the future GM crops with even higher yields and entirely novel GM
varieties of tuber crops should make biofuel production even more efficient and inexpensive.
Genetic modification of crops will not only produce sufficient quantities of tuber crops for food
and animal feed, but will also produce sufficient quantities, which will most likely play a major role
on the biofuel industry. Apart from these, development of suitable improved varieties, increased
content of fermentable sugars (ethanol crops), improved processing characteristics that facilitate
their conversion to biofuels, cultivation technologies for different agro-ecological systems, need
further investigations and the biotechnological tools are likely to play an increasingly important
role in this direction.
5. Conclusion
In the recent years bioethanol produced from tuber crops seems to be a promising aspect, because
of their abundant availability, high starch content and cost effective processing, making it one of
the desirable substrates among all the other substrates such as lignocelluloses and algal biomass,
which have high processing and maintenance cost. The advanced biotechnological tools have
further made it possible to enhance the yield and quality of tuber crops. Besides, the applications
of the new biotechnological tools for processing and producing bioethanol from different tuber
crops are in practice thereby leading to large-scale production of bioethanol. With the technological
development, the rapid increase in the biofuel supply has made the governments to create and
implement new standards so that the supply goes parallel with the demand. In this context, the
governments of different countries support the steady production and use of bioethanol from tuber
crops as an alternative to petroleum fuel. The development of bioethanol production from tuber
crops has been important not only to reduce the fossil fuels dependency but also to lower the
contribution of petroleum derivatives to climate changes and air pollution. There is a continuous
effort on development of novel strain and standardisation of technology for increasing bioethanol
production. Time is not very far when we shall depend on bioethanol as a major source of energy
to meet the crisis of depletion of fossil fuel for transportation and industrial uses.
References
Abubakaret, B. A., M. M. Aliyu, S. A. Mikhail, I. Hamisu, and O. O. Adebayo. 2000. “Phytochemical Screening and
Antibacterial Activities of Vernonia Ambigua. Vernonia blumeoides and Vernoniao ocephala (Asteraceae).” Acta
Poloniae Pharmaceutica – Drug Research 68 (1): 67–73.
Abouzied, M. M., and C. A. Reddy. 1986. “Direct Fermentation of Potato Starch to Ethanol by Cocultures of Aspergillus
niger and Saccharomyces cerevisiae.” Applied and Environment Microbiology 52 (5): 1055–1059.
Abouzied, M. M., and C. A. Reddy. 1987. “Fermentation of Starch to Ethanol by a Complementary Mixture of an
Amylolytic Yeast and Saccharomy cescerevisiae.” Biotechnology Letters 9 (1): 59–62.
Ado, S. A., G. B. Olukotun, J. B. Ameh, and A.Yabaya. 2009. “Bioconversion of Cassava Starch to Ethanol in a Simultane-
ous Saccharification and Fermentation Process by Co-cultures of Aspergillus niger and Saccharomyces cerevisiae.”
The Scientific World Journal 4 (1): 19–22.
Aguilera, R. F. 2008. “How Long Will Petroleum Resources Last?” EU Energy Policy (blog). Accessed July 4.
Akponah, E., and O. O. Akpomie. 2012. “Optimization of Bio-Ethanol Production from Cassava Effluent using
Saccharomyces cerevisiae.” African Journal of Biotechnology 11 (32): 8110–8116.
Alexender, M. A., T. W. Chapman, and T. W. Jeffries. 1989. “Continuous-Culture Responses of Candida shehatae to Shifts
in Temperature and Aeration: Implications for Ethanol Inhibition.” Applied and Environmental Microbiology 55 (9):
2152–2154.
Alvira, P., E. Tomás-Pejó, M. Ballesteros, and M. J. Negro. 2010. “Pretreatment Technologies for an Efficient Bioethanol
Production Process Based on Enzymatic Hydrolysis: A Review.” Bioresource Technology 101 (13): 4851–4861.
Apar, D. K., and B. Ozbek. 2004. “α-Amilase Inactivation During Corn Starch Hydrolysis Process.” Process Biochemestry
39 (12):1877–1892.
Badger, P. C. 2002. “Ethanol from Cellulose: A General Review.” In Trends in New Crops and New Uses, edited by
J. Janick and A. Whipkey, 17–21. Alexandria, VA: American Society for Horticultural Science (ASHS) Press.
Balat, M., and H. Balat. 2009. “Recent Trends in Global Production and Utilization of Bio-Ethanol Fuel.” Applied Energy
86 (11): 2273–2282.
Biofuels factsheet. 2013. Centre for Sustainable Systems, University of Michigan. Pub. No. CSS08 – 09. http://css.snre.
umich.edu/css_doc/CSS08-09.pdf
Brennan, L., and P. Owende. 2010. “Biofuels from Microalgae – a Review of Technologies for Production, Processing,
and Extractions of Biofuels and Co-products.” Renewable and Sustainable Energy Reviews 14 (2): 557–577.
Bro, C., B. Regenberg, J. Forster, and J. Nielsen. 2006. “In silicoaided Metabolic Engineering of Saccharomyces cerevisiae
for Improved Bioethanol Production.” Metabollic Engineering 8 (2): 102–111.
Calinescu, I., P. Chipurici, A. Trifan, and C. Badoiuu. 2012. “Immobilization of Saccharomyces Cerevisiase for the
Production of Bioethanol.” Universitatea Politehnica din Bucuresti Scientific Bulletin, Series B, 74 (1): 33–40.
Campbell, J. E., D. B. Lobell, and C. B. Field. 2009. “Greater Transportation Energy and GHG Offsets from Bioelectricity
Than Ethanol.” Science 324 (5930): 1055–1057.
Castaño, H., M. Cardona, C. Gómez, and A. Acosta. 2011. “Ethanol Production from Cassava Flour in Simultaneous
Enzymatic Hydrolysis and Fermentation System.” Dyna 78 (169): 1–4.
Chandel, A., G. Chandrasekhar, K. Radhika, R. Ravinder, and P. Ravindra. 2011. “Bioconversion of Pentose Sugars into
Ethanol: A Review and Future.” Biotechnology and Molecular Biology Review 6 (1): 8–20.
Chandel, A. K., E. S. Chan, R. Rudravaram, M. L. Narasu, V. Roa, and P. Rabindra. 2007. “Economics and Environmental
Impact of Bioethanol Production Technologies: An Appraisal.” Biotechnology and Molecular Biology Review 2 (1):
014–032.
Chandel, A. K., O. V. Singh, G. Chandrasekhar, L. V. Rao, and M. L. Narasu. 2010. “Key-Drivers Influencing the
Commercialization of Ethanol Based Biorefineries.” Journal Commercial Biotechnology 16: 239–257.
22 H. Thatoi et al.
Chen, Y., R. R. Sharma-Shivappa, D. Keshwani, and C. Chen. 2007. “Potential of Agricultural Residues and Hay for
Bioethanol Production.” Applied Biochemistry and Biotechnology 142 (3): 276–290.
Chen, C. Y., K. L. Yeh, R. Aisyah, D. J. Lee, and J. S. Chang. 2011. “Cultivation, Photobioreactor Design, and Harvesting
of Microalgae for Biodiesel Production: A Critical Review.” Bioresource Technology 102 (1): 71–81.
Cheng, J. J. 2010. Biomass to Renewable Energy Processes. Boca Raton, FL: CRC Press/Taylor & Francis Group.
Cheng, J. J., and R. Timilsina. 2011. “Status and Barriers of Advanced Biofuel Technologies: A Review.” Renewable
Energy 36: 3541–3549.
Dasgupta, D., S. K. Suman, D. Pandey, D. Ghosh, R. Khan, D. Agrawal, R. Kumar, V. T. V. Jain, and D. K. Adhikari.
2013. “Design and Optimization of Ethanol Production from Bagasse Pith Hydrolysate by a Thermotolerant Yeast
Kluyveromyces sp. IIPE453 Using Response Surface Methodology.” Springer Plus 2: 159.
David, L., C. Alice, and N. Ramesh. 2008. “Genetically Engineered Crops for Biofuel Production: Regulatory
Perspectives.” Biotechnology and Genetic Engineering Reviews 25: 331–362.
Delenges, J. P., R. Moletta, and J. M. Navarro. 1996. “Effects of Lignocellulose Degradation Products on Ethanol Fer-
mentations of Glucose and Xylose by Saccharomyces cerevisiae, Zymomonasmobilis, Pichiastipitis and Candida
shehatae.” Enzyme and Microbial Technology 19 (3): 220–225.
Deshpande, V., S. Keskar, C. Mishra, and M. Rao. 1986. “Direct Conversion of Cellulose/Hemicelluloses to Ethanol by
Nuerospora crassa.” Enzyme and Microbial Technology 8 (3): 149–152.
Detroy, R. W., and P. L. Bolen. 1983. “Pan-am. Sypsosium of Fuels and Chemicals.” 3rd, Guatemala 24–26.
Dien, B. S., M. A. Cotta, and T. W. Jeffries. 2003. “Bacteria Engineered for Fuel Ethanol Production Current Status.”
Applied Microbiology and Biotechnology 63 (3): 258–266.
Drapoch. 2008. Biofuel Feed Stock, 69–78. New York: McGraw – Hill Companies Inc.
Duvernay, W. H., M. S. Chinn, and G. C. Yencho. 2013. “Hydrolysis and Fermentation of Sweet Potatoes for Production
of Fermentable Sugars and Ethanol.” Industrial Crops and Products 42: 527–537.
Echegi, U. S. C., P. C. N. Ejikeme, and M. E. Ejikeme. 2013. “Effects of Process Factors on the Synthesis of Bioethanol
from Cassava Tubers using H2 S04 as Catalyst.” The International Journal of Engineering and Science 2 (5):
01–09.
Edem, D. O., C. T. Amugo, and O. U. Eka. 1990. “Chemical Composition of Yam Bean (Sphenostylis sternocarpa).”
Tropical Science 30: 59–63.
Farrell, A. E., R. J. Plevin, B. T. Turner, A. D. Jones, M. O’Hare, and D. M. Kammen. 2006. “Ethanol can Contribute to
Energy and Environmental Goals.” Science 311 (5760): 506–508.
Gksungur, Y., and U. Gven. 1999. “Production of Lactic Acid from Beet Molasses by Calcium Alginate Immobilized
Lactobacillus delbrueckii IFO 3202.” Journal of Chemical Technology and Biotechnology 74 (2): 131–136.
Godoy, A., H. V. Amorim, M. L. Lopes, and A. J. Oliveira. 2008. “Continuous and Batch Fermentation Processes:
Advantages and Disadvantages of These Processes in the Brazilian Ethanol Production.” International Sugar Journal
110 (1311): 175–181.
Graeme, M., and V. Walker. 2010. “Science and Technology of Fuel Alcohol. Bioethanol. Transgenics are Imperative for
Biofuel Crops.” Plant Science 174: 246–263.
Grahovac, J. A., J. M. Dodić, S. N. Dodić, S. D. Popov, D. G. Vuèurović, and A. I. Jokić. 2012. “Future Trends of Bioethanol
Co-production in Serbian Sugar Plants.” Renewable and Sustainable Energy Review 16 (5): 3270–3274.
Hahn-Hägerdal, B., K. Karhumaa, C. Fonseca, I. Spencer-Martins, and M. F. Gorwa Grauslund. 2007. “Towards Industrial
Pentose-Fermenting Yeast Strains.” Applied Microbiology and Biotechnology 74 (5): 937–953.
Han, M., Y. Kim, B. Chung, and G. Choi. 2011. “Bioethanol Production from Optimized Pretreatment of Cassava Stem.”
Korean journal of Chemical Engineering 28 (1): 119–125.
Hashem, M., and S. M. I. Darwish. 2010. “Production of Bioethanol and Associated By-products from Potato Starch
Residue Stream by Saccharomyces cerevisiae.” Biomass Bioenergy 34 (7): 953–959.
Hashem, M., and S. Darwish. 2011. “Production of Bioethanol and Associated By-products from Potato in Very High
Gravity Fermentation.” Biotechnology Letters 29: 233–236.
He, M. X., H. Feng, F. Bai, Y. Li, X. Liu, and Y. Z. Zhang. 2009a. “Direct Production of Ethanol from Raw Sweet
Potato Starch using Genetically Engineered Zymomonas mobilis.” African Journal of Microbiology Research 3 (11):
721–726.
He, M. X.,Y. Li, X. Liu, F. Bai, H. Feng, andY.-Z. Zhang. 2009b. “Ethanol Production by Mixed Culture of Paenibacillus sp
and Zomomonas mobilis using Raw Starchy Material from Sweet Potato.” Annals of Microbiology 59 (4): 749–754.
Hoover, R. 2001. “Composition, Molecular Structure and Physicochemical Properties of Tuber and Root Starches – a
Review.” Carbohydrate Polymers 45 (3): 253–267.
Hyun, H. H., and J. G. Zeikus. 1985. “General Biochemical Characterization of Thermostable Extracellular β-Amylase
from Clostridium Thermosulfurogenes.” Applied and Environmental Microbiology 49 (5): 1162–1167.
Ibeto, C. N., A. U. Ofoefule, and K. E. Agbo. 2011. “A Global Overview of Biomass Potentials for Bioethanol Production:
A Renewable Alternative Fuel.” Trends Applied Science Research 6 (5): 410–425.
Ibrahim, H. A. H. 2012. “Pretreatment of Straw for Bioethanol Prodution.” Energy Procedia 14: 542–551.
IEA. 2011. Analysis of Alternative Transport Fuel Costs Under Different Oil Price Assumptions in the Near and Longer
Term. Paris: OECD/IEA.
Jata, S. K., M. Nedunchezhiyan, T. R. Sahoo, and A. Lenka. 2012. “Sustainable Livelihoods through Tuber Crop.” Odisha
Review 84–90.
Jeon, B. Y., S. J. Kim, D. H. Kim, B. K. Na, D. Y. Park, H. T. Tran, R. Zhang, D. H. Ahn. 2007. “Development of a
Serial Bioreactor System for Direct Ethanol Production from Starch Using Aspergillus niger and Saccharomyces
cerevisiae.” Biotechnology and Bioprocess Engineering 12 (5): 566–573.
Jirasak, K., and S. Buddhiporn. 2011. “Comparative Study of Bioethanol Production from Cassava Peels by Monoculture
and Co-culture of Yeast.” Journal of National Science 45 (2): 268–274.
Kays, S. J., and Y. Wang. 2002. “Sweet-Potato Quality: Its Importance, Assessment and Selection in Breeding Programs.”
Acta Horticulture 583: 187–193.
Khan, R. A., A. Nawaz, M. Ahmed, M. R. Khan, F. Dian, N. Azam, S. Ullah, F. Sadullah, A. Ahmad, M. S. Shah, and N.
Khan. 2012. “Production of Bioethanol through Enzymatic Hydrolysis of Potato.” African Journal of Biotchnology
11 (25): 6739–6743.
Kim, I.-S.,Y.-S. Kim, H. Kim, I. Jin, and H.-S.Yoon, 2013. “Saccharomyces cerevisiae KNU5377 Stress Response During
High-temperature Ethanol Fermentation.” Molecules and Cells 35 (3): 210–218.
Kongkiattikajorn, J., and B. Sornvoraweat. 2011. “Comparative Study of Bioethanol Production from Cassava Peels by
Monoculture and Co-culture of Yeast.” Kasetsart Journal (Natural Science) 45 (2): 268–274.
Kraatz, S. 2008. “Energy Inputs for Corn Production in Wisconsin and Germany.” ASABE Annual International Meeting
Sponsored by ASABE Rhode Island Convention Center Providence, Rhode Island, June 29–July 2.
Kumar, P., D. M. Barrett, M. J. Delwiche, and P. Stroeve. 2009. “Methods for Pretreatment of Lignocellulosic Biomass for
Efficient Hydrolysis and Biofuel Production.” Industrial and Engineering Chemistry Research 48 (8): 3713–3729.
Kymalainen, M. 2007. “Bioethanol – a Possibility in Finland.” In Recent Applications in Bioprocess Engineering., edited
by F. Hameenlinna, H. Kautola, and T. Pirttijarvi, 43–48. London: HAMKin e-julkaisuja7.
Lareo, C., M. D. Ferrari, M. Guigou, L. Fajardo, V. Larnaudie, M. B. Ramírez, and J. Martínez-Garreiro. 2013. “Evaluation
of Sweet Potato for Fuel Bioethanol Production: Hydrolysis and Fermentation.” SpringerPlus 2: 493.
Lee, W. S., I. C. Chen, C. H. Chang, and S. S.Yang. 2012. “Bioethanol Production from Sweet Potato by Co-immobilization
of Saccharolytic Molds and Saccharomyces cerevisiae.” Reneweble Energy 39 (1): 216–222.
Lee O. K., A. L. Kim, D. H. Seong, C. G. Lee,Y. T. Jung, J. W. Lee, and E.Y. Lee. 2013. “Chemo-enzymatic Saccharification
and Bioethanol Fermentation of Lipid – Extracted Residual Biomass of the Microalgae Dunaliella tertiolecta.”
Bioresource Technology 132: 197–201.
Liimatainen, H., T. Kuokkanen, and J. Kaariainen. 2004. “Development of Bio-ethanol Production from Waste Potatoes.”
In Proceedings of the Waste Minimization and Resources Use Optimization Conference, June 10th 2004, University
of Oulu, Finland. edited by E Pongrácz, 123–129. Oulu: Oulu University Press.
Lin, Y., and S. Tanaka. 2006. “Ethanol Fermentation from Biomass Resources: Current State and Prospects. Source: Asian
Centre for Environmental Research, Meisei University, Tokyo, Japan.” Applied Microbiology and Biotechnology 69
(6): 627–642.
Litch, F. O. 2012. Cited in Ethanol Producer Magazine, June 27. http://www.ethanolproducer.com/articles/8907.
Lixin, Z., M. Haibo, S. Liying, and J. Yan. 2007. “Estimation of Un-Used Land Potential for Biofuels Development in
(the) People’s Republic of China.” Applied Energy 86 (1): 52–57.
Lynd, L. R., and M. Q. Wang. 2003. “A Product-Nonspecific Framework for Evaluating the Potential of Biomass-Based
Products to Displace Fossil Fuels.” Journal of Industrial Ecology 7 (3–4): 17–32.
Lynd, L. R., P. J. Weimer, W. H. van Zyl, and I. S. Pretorius. 2002. “Microbial Cellulose Utilization: Fundamentals and
Biotechnology.” Microbioliology and Molecular Biology Review 66 (3): 506–577.
MacLean, H. L., and L. B. Lave. 2004. “Alternative Transport Fuels for the Future.” International Journal of Vehicle
Design 35 (1–2): 27–49.
MarèlneCot, M., M. O. Loret, and J. Francois. 2007. “Physiological Behaviour of Saccharomyces cerevisiae in Aerated
Fed-Batch Fermentation for High Level Production of Bioethanol.” FEMS Yeast Research 7 (1): 22–32.
Marija, B. T., V. K. Budimir, L. L. Miodrag, and B. V. Vlada. 2009. “TheAcid Hydrolysis of Potato Tuber Mash Inbioethanol
Production.” Biochemistry Engineering Journal 43 (2): 208–211.
Mark, C. M., and V. Loosdrecht. 2007. “Mixed Culture Biotechnology for Bioenergy Production.” Current Opinion in
Biotechnology 18 (3): 207–212.
Martín, C.,Y. López,Y. Plasencia, and E. Hernández. 2006. “Characterization of Agricultural and Agro-industrial Residues
as Raw Materials for Ethanol Production.” Chemical and Biochemical Engineering Quarterly 20 (4): 443–447.
McMillan, J. D. 1997. “Bioethanol Production: Status and Prospects.” Renewable Energy 10 (2–3): 295–302.
Millichip, R. J., and H. W. Doelle. 1989. “Large-Scale Ethanol Production from Milo (Sorghum) using Zymomonas
mobilis.” Process Biochemistry 24 (4): 141–145.
MINO, A. K. 2010. “Ethanol Production from Sugarcane in India: Viability, Constraints and Implications.” Submitted in
partial fulfillment of the requirements for the degree of Master of Science in Natural Resources and Environmental
Sciences in the Graduate College of the University of Illinois at Urbana – Champaign, 2010.
Mussatto, S., G. Dragone, P. Guimarães, J. Silva, and L. Carneiro. 2010. “Technological Trends, Global Market, and
Challenges of Bio-Ethanol Production.” Biotechnology Advertisement 28 (6): 817–830.
Narendranath, N. V., K. C. Thomas, and W. M. Ingledew. 2000. “Urea Hydrogen Peroxide Reduces the Number of
Lactobacilli spp., Nourish Yeast, and Leaves No Residues in the Ethanol Fermentation.” Applied and Environmental
Microbiology 66 (10): 4187–4192.
Naylor, R. L., A. J. Liska, M. B. Burke, W. P. Falcon, J. C. Gaskell, S.D. Rozelle, and K.G. Cassman. 2007. “The Ripple
Effect.” Environment 49 (9): 33–46.
Neves, M. A., T. Kimura, N. Shimizu, and K. Shiiba. 2006. “Production of Alcohol by Simultaneous Saccharification and
Fermentation of Low-Grade Wheat Flour.” Brazilian Archives of Biology and Technology 49 (3): 481–490.
Nigam, J. N. 2001. “Ethanol Production from Wheat Straw Hemicelluloses Hydrolysate by Pichia stipitis.” Journal of
Nuwamanya, E., L. Chiwona, R. Kawuki, andY. Baguma. 2012. “Bio-Ethanol Production from Non-Food Parts of Cassava
(Manihot esculenta Crantz).” Ambio 41 (3): 262–270.
24 H. Thatoi et al.
Nweke, F. 2004. New Challenges in the Cassava Transformation in Nigeria and Ghana. Environment and Production
Technology Division. Washington, DC: International Food Policy Research Institute (IFPRI).
Nyerhovwo, J. I. 2004. “Cassava and the Future Biotechnology Issues for Developing Countries”. Electronic Journal of
O’Connor, C. B. 1994. “Rural Dairy Technology. ILRI Training Manual No.1.” International Livestock Center for Africa,
133, Addis Ababa, Ethiopia.
Ogbonna, C. N., and E. C. Okoli. 2010. “Effects of Supplementing Cassava Koji with Crude Amylase Enzymes from
Submerged Cultures on Ethanol Production from Cassava Flour.” Journal of Science and Engineering Technology
17 (3): 9722–9737.
O’Hair, S. K. 1990. “Tropical Root and Tuber Crops.” In Advances in New Crops, edited by J. Janick and J. E. Simon,
424–428. Portland, OR: Timber Press.
Öhgren, K., R. Bura, G. Lesnicki, J. Saddler, and G. Zacchi. 2007. “A Comparison between Simultaneous Saccharification
and Fermentation and Separate Hydrolysis and Fermentation Using Steam-Pretreatment Corn Stover.” Process
Biochemistry 42 (5): 834–839.
Olsson, L., and B. Hahn-Hägerdal. 1996. “Fermentation of Lignocellulosic Hydrolysates for Ethanol Production.” Enzyme
Microbial Technology 18 (5): 312–331.
Osuji, C. M, E. C. Nwanekezi, E. M. Amadi, and C. A. Osuji. 2010. “Yield of Ethanol from Enzyme-Hydrolyzed Yam
(Dioscorea rotundata) and Cocoyam (Xanthosoma sagittifolium) Flours.” Nigerian Food Journal 28 (2): 1–5.
Padmaja, G., M. George, and S. N. Moorthy. 2006. “Detoxification of Cassava During Fermentation with a Mixed Culture
Innoculumns.” Journal of the science of food and Agriculture 63 (4): 473–481.
Palmqvist, E., and B. Hahn-Hägerdal. 2000. “Fermentation of Lignocellulosic Hydrolysates. I: Inhibition and Detoxifi-
cation.” Bioresource Technology 74 (1): 17–24.
Pasha, C., R. C. Kuhad, and L.V. Rao. 2007. “Strain Improvement of Thermotolerant Saccharomyces cerevisiae VS3
Strain for Better Utilization of Lignocellulosic Substrates.” Journal of Applied Microbiology 103 (5): 1480–1489.
Patni, N., G. S. Pillai, and A. H. Dwivedi. 2011. Analysis of Current Scenario of Biofuels in India Specifically Bio-Diesel
and Bio-Ethanol, 382–481. Ahmedabad: Institute of Technology, Nirma University.
Pearlman, D. 1950. “Observations on the Production of Ethanol by Fungi and Yeast.” American Journal of Botany 37:
237–224.
Plessas, S., A. Bekatorou, A. A. Koutinas, M. Soupioni, I. M. Banat, and R. Marchant. 2007. “Use of Saccharomyces
cerevisiae Cells Immobilized on Orange Peel as Biocatalyst for Alcoholic Fermentation.” Bioresource Technology
98 (4): 860–865.
Prasad, S., A. Singh, and H. C. Joshia. 2007. “Ethanol as an Alternative Fuel from Agricultural, Industrial and Urban
Residues.” Resource Conservation Recycling 50 (1): 1–39.
Rani, P., S. Sharma, F. C. Garg, K. Raj, and L. Wati. 2010. “Ethanol Production from Potato Flour by Saccharomyces
cerevisiae.” Indian Journal of Science and Technology 3 (7): 733–736.
Ray, R. C., and S. N. Moorthy. 2007. “Exopollysaccaride Production from Cassava Starch Residue by Auribassadium
pullulans Strain MTCC 7521.” Journal of Scientific and Industrial Research 66 (3): 252–255.
Reddy, O. V. S., and S. C. Basappa. 1996. “Direct Fermentation of Cassava Starch to Ethanol by Mixed Cultures of
Endomycopsis fibuligera and Zymomonas mobilis: Synergism and limitations.” Biotechnology Letters 18 (11): 1315–
1318.
Rosenberg, S. L. 1980. “Fermentation of Pentose Sugars to Ethanol and Other Neutral Products by Microorganisms.”
Enzyme Microbial Technology 2 (3): 185–193.
Sánchez, O., and C. Cardona. 2008. “Trend in Biotechnological Production of Fuel Ethanol from Different Feedstock.”
Bioresource Technolology 99 (13): 5270–5295.
Scott, G. J., and L. Maldonado. 1997. Sweet Potato for the New Millenium: Trends in Production and Utilization in
Developing Countries. CIP Program Report 329–335.
Scott, G., R. Best, M. W. Rosegrant, and M. Bokanga. 2000. “Roots and Tubers in the Global Food System – A Vision
Statement to the Year 2020.” CIAT-CIP-IFPRI-IITA-IPGRI-CIP: 45, Lima, Peru.
Senthilkumar, V., and P. Gunasekaran. 2005. “Bioethanol Production from Cellulosic Substrates: Engineered Bacteria and
Process Integration Challanges.” Journal of Scientific and Industrial Research 64 (11): 845–853.
Shama, G. 1988. “Developments in Bioreactors for Fuel Ethanol Production.” Process Biochemistry 23: 138–145.
Shanavas, S., G. Padmaja, S. N. Moorthy, M. S. Sajeev, and J. T. Sheriff. 2011. “Process Optimization for Bioethanol
Production from Cassava Starch Using Novel Eco-friendly Enzymes.” Biomass and Bioenergy 35 (2): 901–909.
Singh, A., P. K. R. Kumar, and K. Schugerl. 1992. “Bioconversion of Cellulosic Materials to Ethanol by Filamentous
Fungi.” Advances in Biochemical and Biotechnology 45: 29–55.
Skoog, K., and B. Hahn-Hägerdal. 1988. “Xylose Fermentation.” Enzyme Microbial Technology 10: 66–80.
Sommer, P., T. Georgieva, and B. K. Ahring. 2004. “Potential for using Thermophilic Anaerobic Bacteria for Bioethanol
Production from Hemicellulose.” Biochemical Society Transactions 32 (2): 283–289.
Sonneitner, B. 1983. “Biotechnology of Thermophilic Bacteria Growth, Products and Application.” Advances in
Biochemical Engineering/Biotechnology 28: 69–138.
Sovorawet, B., and J. Kongkiattikajorn. 2012. “Bioproduction of Ethanol in SHF and SSF from Cassava Stalks.” KKU
Research Journal 17 (4): 565–572.
Srichuwong, S., M. Fujiwara, X. Wang, T. Seyama, R. Shiroma, M. Arakane, N. Mukojima, and K. Tokuyasu. 2009.
“Simultaneous Saccharification and Fermentation (SSF) of Very High Gravity (VHG) Potato Mash for the Production
of Ethanol.” Biomass Bioenergy 33 (5): 890–898.
Srinivas, T. 2007. “Industrial Demand for Cassava Starch in India.” Sarch/Starke 59 (10): 477–481.
Sun, Y., and J. Y. Cheng. 2002. “Hydrolysis of Lignocellulosic Materials for Ethanol Production: A Review.” Bioresource
Technology 83 (1): 1–11.
Swaider, J., and G. Ware. 2002. Producing Vegetable Crops. 5th ed., Vol. 22, 569–578. Danville, IL: Interstate Publishers.
Swain, M. R., J. Mishra, and H. N. Thatoi. 2013. “Bioethanol Production from Sweet Potato (Ipomoea batatasL.) Flour
Using Co-culture of Trichoderma sp. and Saccharomyces cerevisiae in Solid-State Fermentation.” Brazzilian Archives
Biology and Technology 56 (2): 171–179.
Szambelan, K., J. Nowak, and Z. Czarnecki. 2004. “Use of Zymomonas mobilis and Saccharomyces cerevisiae mixed with
Kluyveromyces fragilis for Improved Ethanol Production from Jerusalem Artichoke Tubers.” Biotechnology Letters
26 (10): 845–848.
Takahashi, T., H. Shimoi, and K. Ito. 2001. “Identification of Genes Required for Growth under Ethanol Stress Using
Transposon Mutagenesis in Saccharomyces cerevisiae.” Molecular Genetics and Genomics 265 (6): 1112–1116.
Takashi, W., S. Sathaporn, S. Mitsuhiro, S. Arakane, M. Tamiya, I. Yoshinaga, M. Watanabe, A. Yamamoto, K. Ando,
T. Tokuyasu, and S. Nakamura. 2010. “Selection of Stress-Tolerant Yeasts for Simultaneous Saccharification and
Fermentation (SSF) of Very High Gravity (VHG) Potato Mash to Ethanol.” Bioresource Technology 101 (24):
9710–9714.
Teramoto, Y., S. H. Lee, and H. Endo. 2008. “Pretreatment of Woody and Herbaceous Biomass for Enzymatic
Saccharification Using Sulfuricacid-Free Ethanol Cooking.” Bioresource Technology 96 (18): 8856–8863.
Tokgoz, S. 2008. “The Impact of Energy Markets on the EU Agricultural Sector.” Proceedings of the 12th congress of the
European association of agricultural economists – EAAE 2008, Ghent, Belgium, August 26–29.
Tran, A. V., and R. P. Chambers. 1986. “Red Oak Wood Derived Inhibitors in the Ethanol Fermentation of Xylose by
Pichia stipitis CBS 5776.” Biotechnology Letters 7 (11): 841–846.
USDA GAIN, Biofuels Annual China. 2012. Peoples Republic of China. Report Number: 12044. http://gain.fas.usda.gov/
Recent%20GAIN%20Publications/Biofuels%20Annual_The%20Hague_EU-27_8-13-2013.pdf
USDA GAIN, Biofuels Annual India. 2012. Report Number: IN2081. http://gain.fas.usda.gov/Recent%20GAIN
%20Publications/Biofuels%20Annual_New%20Delhi_India_6-20-2012.pdf
Van Voorst, F., J. Houghton-Larsen, L. Jonson, M. C. Kielland-Brandt, and A. Brandt. 2006. “Genome-Wide Identification
of Genes Required for Growth of Saccharomyces cerevisiae Under Ethanol Stress.” Yeast 23 (5): 351–359.
Vijaya, O., R. Sarathi, and S. C. Basappa. 1996. “Direct Fermentation of Cassava Starch to Ethanol by Mixed Cultures
of Endomycopsis fibuligera and Zymomonas mobilis: Synergism and Limitations.” Biotechnology Letters 18 (11):
1315–1318.
Wagner, W. 2005. “Biodiesel and Ethanol from Soybeans and Sugar Beets.” BioCycle 46 (6): 10.
Wang, F. Q., C. J. Gao, C. Y. Yang, and P. Xu. 2007. “Optimization of an Ethanol Production Medium in Very High Gravity
Fermentation.” Biotechnology Letters 29 (2): 233–236.
Wang, G. S., X. J. Pan, J. Y. Zhu, R. Gleisner, and D. Rockwood. 2009. “Sulfite Pretreatment to Overcome Ecalcitrance
of Lignocellulose (SPORL) for Robust Enzymatic Saccharification of Hardwoods.” Biotechnology Progress 25 (4):
1086–1093.
Wen, S., I. Lee, C. Chen, C. Hsiung, C. Shang, and S. Yang. 2002. “Bioethanol Production from Sweet Potato by Co-
immobilization of Saccharolytic Molds and Saccharomyces cerevisiae.” Applied and Environmental Microbiology
39 (1): 540–564.
Wheals, A. E., L. C. Basso, D. M. G. Alves, and H. V. Amorim. 1999. “Fuel Ethanol after 25Years.” Trends in Biotechnology
17 (12): 482–487.
William, H., S. Mari, G. Chinn, and Y. Craig. 2013. “Hydrolysis and Fermentation of Sweet Potatoes for Production of
Fermentable Sugars and Ethanol.” Industrial Crops and Products 42: 527–537.
Williams, L. A., and G. Ziobro. 1982. “Processing and Fermentation of Jerusalem Artichoke for Ethanol Production.”
Biotechnology Letters 4 (1): 45–50.
Wyman, C. E., B. E. Dale, R. T. Elander, M. Holtzapple, M. R. Ladisch, and Y. Y. Lee. 2005. “Coordinated Development
of Leading Biomass Pretreatment Technologies.” Bioresource Technology 96 (18): 1959–1966.
Xiros, H., and P. Christakopoulos. 2009. “Enhanced Ethanol Production from Brewer’s Spent Grain by a Fusariumoxys-
porum Consolidated System.” Biotechnology and Biofuels 21: 421–432.
Xiu, Z. L., and A. P. Zeng. 2008. “Present State and Perspective of Downstream Processing of Biologically Produced 1,
3-Propanediol and 2,3-Butanediol.” Applied Microbiology and Biotechnology 78 (6): 917–926.
Yang, B., and C. E. Wyman. 2008. “Pretreatment: The Key to Unlocking Low Cost Cellulosic Ethanol.” Biofuels,
Bioprodoucts and Biorefining 2 (1): 26–40.
Yang, F., M. Cao, Y. Jin, X. Yang, and S. Tian. 2012. “Development and Application of Saccharomyces Cerevisiae
Cell-surface Display for Bioethanol production.” Chinese Journal of Biotechnology 28 (8): 901–911.
Yoon, J. J., Y. J. Kim, S. H. Kim, H. J. Ryu, J. Y. Choi, G. S. Kim, and M. K. Shin. 2010. “Production of Polysaccharides
and Corresponding Sugars from Red Seaweed.” Advances in Material Research 93–94: 463–466.
Yuwa-Amornpitak, T. 2010. “Ethanol Production from Cassava Starch by Selected Fungi from TanKoji and Saccharomyces
cerevisiae.” Biotechnology 9 (1): 84–88.
Zhang, M., C. Eddy, K. Deanda, M. Finkelstein, and S. Picataggio. 1995. “Metabolic Engineering of a Pentose Metabolism
Pathway in Ethanologenic Zymomonas mobilis.” Science 267 (5195): 240–243.
Zhang, L., H. Zhao, M. Gan, Y. Jin, X. Gao, Q. Chen, J. Guan, and Z. Wang. 2011. “Application of Simultaneous
Saccharification and Fermentation (SSF) from Viscosity Reducing of Raw Sweet Potato for Bioethanol Production
at Laboratory, Pilot and Industrial Scales.” Bioresource Technology 102 (6): 4573–4579.
26 H. Thatoi et al.
Zhang, Y., M. Zheng, H. Xiong, F. Hong, B. Fan, L. Yi, and L. Xun. 2009. “Direct Production of Ethanol from Raw
Sweet Potato Starch Using Genetically Engineered Zymomonas mobilis.” African Journal of Microbiology 3 (11):
721–726.
Zhang, Y., Y. Zhu, and Y. Li. 2009. “The Importance of Engineering Physiological Functionality into Microbes.” Trends
in Biotechnology 27 (12): 664–672.
Zhang, L., H. Zhao, M. Gan, Y. Jin, X. Gao, Q. Chen, J. Guan, and Z. Wang. 2011. “Application of Simultaneous
Saccharification and Fermentation (SSF) from Viscosity Reducing of Raw Sweet Potato for Bioethanol Production
at Laboratory, Pilot and Industrial Scales.” Bioresource Technology 102 (6): 4573–4579.
Zhao, X. Q., and F. W. Bai. 2009. “Yeast Flocculation: New Story in Fuel Ethanol Production.” Biotechnology
Advertisement 27 (6): 849–856.
Zhao, X., K. Cheng, and D. Liu. 2009. “Organosolv Pretreatment of Lignocellulosic Biomass for Enzymatic Hydrolysis.”
Applied Microbiology and Biotechnology 82 (5): 815–827.
Zhong, W., S. Zhang, and Y. Zheng. 2006. “Cloning and Expression of any E Gene from Bacillus subtilis in Zymomonas
mobilis and Direct Production of Ethanol from Soluble Starch.” Biotechnology and Bioprocess Engineering 17 (4):
780–786.
Ziska, L. H., G. B. Runionb, M. Tomeceka, S. A. Priorb, H. A. Torbetb, and R. Sichera. 2009. “An Evaluation of Cassava,
Sweet Potato and Field Corn as Potential Carbohydrate Sources for Bioethanol Production inAlabama and Maryland.”
Biomass and Bioenergy 33 (11): 1503–1508.

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On: 04 May 2015, At: 03:16

International Journal of Sustainable

Bioethanol production from tuber

To link to this article: http://dx.doi.org/10.1080/14786451.2014.918616

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Bioethanol production from tuber crops using fermentation

(Received 6 December 2013; final version received 19 April 2014)

Keywords: tuber crops; micro-organisms; fermentation techniques; bioethanol; biotechnological tools

∗ Corresponding author. Email: hn_thatoi@rediffmail.com

© 2014 Taylor & Francis

Table 1. Worldwide bioethanol production from different feedstocks.

Leading ethanol producing regions worldwide

Location Feedstock 2007 2008 2009 2010 2011 2012

transportation fuel burning than non-renewable gasoline, as it is more oxygenated (Wheals

Figure 1. World bioethanol production.

Figure 2. Biomass for different generation bioethanol production.

2. Types of tuber crops

Table 2. Bioethanol production potential of different tuber

Tuber crops and Ethanol

Cassava 4711 Drapoch (2008)

Table 3. Chemical composition of different tuber crops used in bioethanol production.

Tuber Moisture Protein Crude Nitrogen free Vitamin Starch

Cassava 59.4 0.7 0.6 34.1 25.2 20–25

Tanzania, Cuba, Pradesh, Tamil

2.2. Sweet potato

2.5. Sugar beet

2.7. Jerusalem artichoke

Jerusalem artichoke (Helianthus tuberosus) is a tuberous-rooted perennial crop belongs to the

2.8. Asphodelus aestivus

3. Bioethanol production technology

3.1. Different feedstocks for fermentation

3.1.1. Sucrose-containing feedstock

3.1.2. Starchy materials

3.1.3. Lignocellulose biomass

3.2. Micro-organisms in bioethanol fermentation

Ethanol fermentation is a biological process in which sugar (hexose) is converted to ethanol by

3.3. Culture types used in fermentation

Organisms Substrate Treatment Reference

Cassava root peels Enzymatic hydrolysis Akponah and Akpomie (2012)

Organisms Substrate Treatment Reference

or covalent), entrapment in polymeric matrices or retention by membranes has been successful

3.4. Fermentation techniques

3.5. Process development

Process development is an amalgamation of different operations for a particular process that

Drying and milling

3.5.2. Enzymatic hydrolysis

3.5.3. Chemical hydrolysis

Conversion of starchy materials to glucose monomers by saccharification process can be achieved

3.5.4. Fermentation and distillation

4. Biotechnological tools for bioethanol production from tuber crops

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