3

Sorghum as a Multifunctional Crop

for the Production of Fuel Ethanol:
Current Status and Future Trends
Sergio O. Serna-Saldívar*, Cristina Chuck-Hernández,
Esther Pérez-Carrillo and Erick Heredia-Olea
Departamento de Biotecnología e Ingeniería de Alimentos,
Centro de Biotecnología. Tecnológico de Monterrey, Monterrey, N. L.
México

1. Introduction
Nowadays, there is a growing interest for alternative energy sources because of the
reduction of fossil fuel production. Ethanol used as automotive fuel has increased at least six
times in the current century. According to the Renewable Fuels Association, in 2010 the USA
bio-refineries generated 13 billion gallons of fuel ethanol and the year before worldwide
production reached 19 billion. This noteworthy increment is in its majority based on maize
and sugar cane as raw materials (Berg, 2004; Renewable Fuels Association, 2010). The use of
these feedstocks has triggered concerns related to food security especially today when the
world population has reached 7 billion people.
The relatively sudden rise in food prices during 2008, 2010 and 2011 has been attributed
mainly to the use of maize for bioethanol even when other factors like droughts or changes
in global consumption patterns have also played a major role (World Food Program, 2008).
Food price projections indicate that this situation will worsen, breaking the downward trend
registered in food prices in the last thirty years (The Economist, 2007).
Even if there was not a food-fuel controversy especially due to the current conversion of
millions of tons of maize for bioethanol, the use of only this crop cannot support the
ambitious objectives of renewable fuel legislation in countries like the United States of
America, where a target of 36 billion gallons of liquid biofuels have been established for
2022. In order to meet this requirement all the 333 million tons of maize yearly produced by
USA should be channelled to biorefineries. This production represents 2 and 16 times the
maize harvested in countries like China and Mexico respectively, which in turn are two of
the five top world producers.
Environmental factors have been also pushing for the quest of new crops dedicated
exclusively for liquid automotive fuel in order to reduce the use of prime farming land,
irrigation water and other resources. A dedicated energy crop ideally must meet several
requirements such as: high biomass yield and growth rate, perennial, with reduced input
necessities, fully adapted to the geographic regions where will be planted, easy to
manipulate via genetic improvement, non-invasive, tolerant to stress and with a good
carbon sequestration rate among others (Jessup, 2009). At the present time, energy crops are

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52 Bioethanol

mainly represented by perennial grasses as switchgrass (Panicum virgatum L.), energy cane
(Saccharum spp), sweet and forage sorghum (Sorghum bicolor), miscanthus (Miscanthus spp.)
as well as other short-rotation forest resources (willow –Salix spp- and poplar –Populus spp)
(Jessup, 2009; McCutchen et al., 2008).
The development of new and improved enzymes, bioprocesses and feedstocks could lead to
cost reduction from an estimated of 0.69 cents to below 0.51 cents/L that nowadays is the
benchmark established for starchy raw materials (Kim & Day, 2011). Besides the
development of dedicated crops for energy, one of the best approaches for cost reduction
and optimal use of resources is the use of flexible facilities allowing the integration of
different streams of same or different feedstocks. Flexibility, balance, diversification and
regionalization are indeed keywords in the development of solutions to meet future world
energy demands.
In tropical, subtropical, and arid regions from the United States, Mexico, China, India,
Southern Africa, and other developing countries, where agronomic harsh conditions prevail,
one of the most promising crops for fuel is sorghum (Sorghum bicolor (L.) Moench) (Reddy et
al., 2005; Zhang et al., 2010). This is a high efficient photosynthetic crop that reached a
worldwide production of 56 million tons of grain in 2009 (FAOSTAT, 2011), just behind
maize, wheat, rice and barley. Almost 30% of this production is harvested in North America
where sorghum is mainly used for feed. Sorghum is a C4 plant, highly resistant to biotic and
abiotic factors as insects, drought, salinity, and soil alkalinity. Furthermore, this crop has
one of the best rates of carbon assimilation (50 g/m2/day) which in turn allows a fast
growth and a better rate of net CO2 use (Prasad et al., 2007). Sorghum requires one third of
the water with respect to sugar cane and 80 to 90% compared to maize (Almodares & Hadi,
2009; Wu et al., 2010b). Thus, sorghum is considered as one of the most drought resistant
crops. Furthermore, sorghum requires approximately one third of the fertilizer required by
sugar cane (Kim & Day, 2011) and its growth cycle is between 3 to 5 months allowing two or
three crops per year instead of one commonly obtained with sugarcane. Besides
environmental advantages, sorghum is one of the more acquiescent plants to genetic
modification because is highly variable in terms of genetic resources and germplasm. This
facilitates plant breeding and development of new cultivars adapted to different regions
around the globe (Zhang et al., 2010).
Sorghum can be classified in four broad groups: grain, sweet, forage and high biomass. All
belong basically to the same species and virtually there are no biological or taxonomic
differences (Wang et al., 2009). Grain sorghum is used mainly as food, feed and for starch
production. In the United States only a small percentage of fuel ethanol (around 2-3%) is
obtained from grain sorghum (Renewable Fuels Association, 2010; Turhollow et al., 2010;
Zhao et al., 2008), but in 2009 about 30% of the U.S. grain sorghum crop was used for
ethanol production (Blake, 2010).
On the other hand, forage sorghum is characterized as a high biomass crop. This capacity
has been boosted by intensive research programs worldwide, focused in the design of
new varieties tailored for ethanol production (Rooney et al., 2007). The main product
obtained from sweet sorghums is the fermentable sugar rich juice that is produced and
accumulated in the stalks in a similar fashion as sugar cane. The extracted sweet juice is
mainly composed of sucrose, glucose, and fructose, and thus can be directly fermented
into ethanol with efficiencies of more than 90% (Wu et al., 2010b). According to
Almodares & Hadi (2009) sorghum yields a better energy output/input ratio compared to
other feedstocks such as sugar cane, sugar beet, maize and wheat. Altogether with the

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Sorghum as a Multifunctional Crop
for the Production of Fuel Ethanol: Current Status and Future Trends 53

juice, the residue or bagasse can be also converted to ethanol or used for other traditional
applications.
In summary, sorghum is a crop well adapted to adverse climatic conditions which at this
time is one of the growing concerns in agronomic projections. This is mainly due to the
change of rain patterns and climate, greenhouse effect and the steadily rise of world
temperature. Given all these advantages of sorghum as a potential source of biofuels, the
objective of this chapter is to explore its potential, as an integrated crop for fuel production
in terms of yield and technologies available for processing. The chapter especially focuses on
optimum technologies to produce bioethanol from sweet sorghums, starchy grains and
biomass from dedicated crops.

2. Botanical features and agronomic characteristics

Sorghum is a member of Poaceae family, a high-efficient photosynthetic crop, well adapted
to tropical and arid climates. As a result, sorghum is extremely efficient in the use of water,
carbon dioxide, nutrients and solar light (Kundiyana, 1996; Serna-Saldívar, 2010). This crop
is considered one of the most drought resistant, making it one of the most successful in
semi-desert regions from Africa and Asia (Woods, 2000). This resistance is due mainly to its
photosynthetic C4 metabolism that allows sorghum to accumulate CO2 during the night, to
lower the photorespiration rate in presence of light, to reduce the loss of water across the
stoma and the waste of carbon (Keeley & Rundel, 2003).
The leaves of sorghum and maize are similar but in the case of sorghum they are covered by
a waxy coat that protects the plant from prolonged droughts. The sorghum grain is grouped
in panicles and the plant height ranges from 120 to 400 cm depending on type of cultivar
and growing conditions. An advantage of sorghum compared to maize is that it has a
comparatively lower seed requirement because only 10 to 15 kg/ha are used compared with
40 kg/ha required by other cereals (Kundiyana, 1996). In some regions is possible to
produce multiple crops per year, either from seed (replanting) or from ratoon (Saballos,
2008; Turhollow et al., 2010).

3. Chemical composition
3.1 Juice from sweet sorghum
The mature stems of sweet sorghum contain about 73% moisture and the solids are divided
in structural and non-structural carbohydrates. Approximately 13% are non-structural
carbohydrates composed of sucrose, glucose and fructose, in variable amounts according to
cultivar, harvesting season, maturity stage, among other agronomic factors (Mamma et al.,
1996; Phowchinda et al., 1997). Anglani (1998) suggests a classification of sweet sorghums
based on proportion of soluble sugars in the juice. The first group with a high content of
sucrose (sugary type) and the second with more monosaccharides (glucose and fructose)
compared to other soluble carbohydrates (syrup type). Smith et al. (1987) in their evaluation
of six sweet sorghum varieties throughout four years in nine different locations did not find
significant differences in sugar content or composition. The typical composition indicates
that around 70% was sucrose and the rest glucose and fructose in equal parts. In stem dry
basis, Woods (2000) reported fermentable sugars content between 41 to 44% in Keller and
Wray varieties with 80 and 63% represented by sucrose and the rest by glucose and fructose.
A fiber variety analyzed by the same author (H173) reached only 20% fermentable sugars
based on the dry stem weight; sucrose, glucose and fructose were found in equivalent

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amounts (around 7% for each sugar). Compared to sugar cane, the main difference is that
the sucrose content in cane is significantly higher compared to glucose and fructose (90, 4
and 6%respectively) and the total content sugar is 49% of the dry stem weight. In general
terms, composition of simple sugars in sweet sorghum juice is 53-85, 9-33 and 6-21% for
sucrose, glucose and fructose, respectively (Gnansounou et al., 2005; Mamma et al., 1996;
Phowchinda et al., 1997; Prasad et al., 2007).
Beyond the proportion of soluble sugars in sweet sorghum plants, the yield of total sugars
per harvested area is a better guide in the analysis for fuel production. Woods (2000)
reported for sweet sorghum cultivars (Keller, Wray and H173) an average of 7, 10 and 4 ton
of fermentable sugars/ha respectively, significantly lower compared to the 17 ton/ha for
sugarcane indicated by the same author. The varieties studied by Davila-Gomez et al. (2011)
yielded an average of 1.85 to 2.03 ton of sugar/ha, whereas Smith et al. (1987) in a extensive
study performed in several locations of continental United States and Hawaii, obtained from
4.5 to 10.6 ton/ha. In other varieties evaluated in China, the best yields reached 18 ton/ha
(Zhang et al., 2010).
Sugars in sweet sorghum are very sensitive to microbial contamination especially after
crushing stalks for juice production. In data reported by Davila-Gomez et al. (2011), the
percentage of sugars, as °Brix before fermentation, was lower (11 to 24% lower) than the
obtained immediately after harvest in summer time, when temperatures easily reached 32°C
in Northeast Mexico. The microbial contamination was the most obvious explanation of this
phenomenon. Besides, the sucrose proportion in the fermented juices was lower in relation
to glucose and fructose (0 to 10% of total). This can be related to invertase activity of
contaminating wild yeasts that hydrolyzed sucrose into glucose and fructose. These
monomers are quickly metabolized by means of facilitated diffusion into the yeast cell. Wu
et al. (2010b), working with cultivars with 16 to 18% of fermentable sugars, found that as
much as 20% of substrate can be lost in 3 days at 25°C. This loss corresponds to
approximately 700 L ethanol/ha when a yield of 50 ton of sorghum stems/ha is considered.
Daeschel et al. (1981) reported that juices can be preserved during 14 days at 4°C without
detectable changes or deterioration (sour odor and foaming). These authors also reported
that the dominant spoilage microorganisms were Leuconostoc mesenteroides and Lactobacillus
plantarum at 25 and 32°C, respectively and recommended to process the juice within five
hours after extraction.

3.2 Sorghum grain

Sorghum grain is a naked caryopsis composed of three major anatomical parts: pericarp,
germ, and endosperm. The pericarp is composed of epicarp, mesorcarp and endocarp
(cross and tube cells). Among cereals, sorghum is the only one that can contain significant
amounts of starch granules in the mesocarp cells. The starch-devoid germ is rich in fat,
soluble sugars and proteins (albumins and globulins) whereas the endosperm is divided
into the single layered aleurone and the starchy endosperm cells positioned in the
corneous and floury or chalky regions of the endosperm. The endosperm constitutes the
largest fraction of the kernel and where almost all the starch is contained. Similar to
maize, sorghum contains 60 to 70% of starch. The endosperm texture and hardness are
highly related to the performance of the grain during several stages of ethanol production.
In general terms, composition of sorghum is similar to maize with a few small but
significant differences mainly in protein and fat concentrations. Sorghum for instance, has
an average 1% less fat and 1.5 to 2.0% more crude protein compared to maize. Both

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Sorghum as a Multifunctional Crop
for the Production of Fuel Ethanol: Current Status and Future Trends 55

sorghum and maize have more than 50% of this protein as prolamins named kafirins and
zeins, respectively. In sorghum, approximately half of the prolamin fraction is bound. In
contrast, approximately 70% of the maize prolamins are free or alcohol-soluble. There are
some sorghum varieties that contain significant amounts of condensed tannins in the
testa. These sorghums are classed as type III and have a lower nutritional value compared
to other sorghums and maize. This is due to the presence of tannins that bind proteins and
inactivate enzymes. As a result, high tannin sorghums may have reduced ethanol yields
(Serna-Saldivar, 2010).
One of the most noteworthy differences between sorghum and maize is its starch granule-
protein matrix interaction that negatively affects the susceptibility of both proteins and
starch to enzyme hydrolyses. These structural differences affect protein digestibility and
the speed of dextrins and glucose production during liquefaction and saccharification and
thereafter the efficiency of yeast fermentation. Kafirins, despite the high sequence
homology with zeins, tend to be less digestible especially after wet-cooking indicating the
change in conformational structure attributed to formation of disulphide bonds. This is
due to its high hydrophobicity which also makes possible the formation of additional
protein aggregates that enhance the formation of more covalent bonds compared to zeins
(Wong et al., 2009). Prolamins in the kernel are concentrated in protein bodies arranged
among starch granules. The protein body composition in maize and sorghum is also
similar, with alpha kafirin in the inner core surrounded by beta and gamma kafirins. The
difference with maize is that during wet thermal processes the external part of protein
body seems to form a net that makes difficult to access the alpha portion that is in turn
more digestible than the beta and gamma counterparts. This phenomenon affects starch
digestibility because in sorghum is 15 to 25% less digestible compared to maize. Taylor &
Belton (2002) indicate that in sorghum, a complex rather than a simple obstruction
mechanism between kafirins and starch is more likely to occur. This is the main reason
why sorghum has lower susceptibility to hydrolysis and fermentation and yields less fuel
ethanol compared to maize. Besides the starch-protein relationship, some other factors
such as mash viscosity, amount of phenolic compounds, ratio of amylose to amylopectin
and formation of amylose-lipid complex in the mash, limit the rate of enzymatic
hydrolysis and fermentation efficiency during bioethanol production. For instance, starch
in amylose-lipid complex cannot be converted into fermentable sugars, reducing
conversion rate and final ethanol yield (Wang et al., 2008).

3.3 Sorghum bagasse and straw

As stated in section 3.1, besides water-soluble sugars (sucrose, glucose and fructose),
sorghum is composed by structural cell wall carbohydrates primarily cellulose and
hemicellulose, which in turn can be hydrolyzed and used as substrate for ethanol
production (Sipos et al., 2009).
Sorghum bagasse is the residual fraction obtained after juice extraction from sweet sorghum
whereas sorghum straw is the remaining material usually left on the field after threshing.
The composition and proportion of fibrous-structural fractions in sorghum is widely
reported and varies according to intrinsic and extrinsic factors such as cultivar type,
maturity and climatic conditions. An average of 15% of the total weight corresponds to the
fibrous portion within a range from 12 to 17% (Woods, 2000).
In sweet sorghum bagasse, average content of cellulose, hemicelluloses and lignin is 34-44%,
27-25%, and 18-20% respectively (Ballesteros et al., 2003; Kim & Day, 2011; Sipos et al., 2009).

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56 Bioethanol

Table 1 summarizes chemical composition of sweet sorghum bagasse and straw compared
to energy-dedicated sugar cane, maize, wheat and rice counterparts.

Feedstock Fiber(%) Cellulose(%) Hemicellulose (%) Lignin (%) Ash (%)

Sweet sorghum 13.0 44.6 27.1 20.7 0.4
Sweet sorghum 2 - 25.0 22.0 4.0 -
Sweet sorghum
- 41.3 24.6 14.0 3.7
bagasse3
Sorghum straw - 32.4 27.0 7.0 0.7
Sugar cane 13.5 41.6 25.1 20.3 4.8
Energy cane 26.7 43.3 23.8 21.7 0.8
Corn stover - 40.0 28.0 21.0 7.0
Wheat straw - 38.0 32.0 19.0 8.0
Rice straw - 36.0 28.0 14.0 20.0
1Modified from Kim & Day (2011) and Reddy & Yang (2005). All data expressed in dry weight basis.
Percentage of fiber is based in 100% of original material and cellulose, hemicellulose, lignin and ash are
percentages of the total fiber; 2Wray variety (Woods, 2000); 3 Data yet not published from sweet
sorghum bagasse harvested in Central Mexico and manually pressed for juice extraction.
Table 1. Fiber composition of different ethanol feedstock 1

4. Ethanol fuel from sweet sorghum juice

Sweet sorghum juice can be used for syrup, molasses, sugar and ethanol production with
average fermentation efficiencies from 85 to 90% (Almodares & Hadi, 2009; Prasad et al.,
2007; Wang et al., 2009; Wu et al., 2010b). The sweet sorghum juice is not commonly used for
crystallized sugar production because of the presence of significant amounts of inverted
sugars (glucose and fructose) that makes difficult crystallization in large-scale processes.
However, the sweet sorghum juice, rich in fermentable sugars, has an excellent potential for
yeast fermentation (Turhollow et al., 2010; Woods, 2000).
The sweet sorghum juice is obtained through a mechanical operation with a roller mill
composed by a set of cylinders, similar to the ones employed by the sugar cane mills. Water
is added during the last stage of the crushing process with the aim to augment the
solubilization of residual sugars associated to the bagasse. The sweet sorghum juice yields
around 50% in relation to the initial weight of the stems (Wu et al., 2010b). However, these
authors describe an extraction process by pressing, which results in lower yields compared
to roller mills. Furthermore, pressing is a batch process which is difficult to optimize for
industrial conditions.
Approximately 90% of fermentable sugars from sorghum stalks can be obtained after
conventional roller-milling, yielding an extraction ratio of 0.7 in relation to the initial plant
weight (Almodares & Hadi, 2009). Gnansounou et al. (2005) reported extraction ratios
ranging from 0.59 to 0.65 for the sweet sorghum cultivars Kelley, Wray, Río and Tianza. On
the other hand, Kundiyana (1996) observed that extraction percentages varied between 47 to
58%, close to values observed by our research group in central Mexico (unpublished data).
After extraction, the sweet sorghum juice is fermented, distilled and the ethanol finally
dehydrated (Fig. 1). This is the simplest way to produce fuel ethanol because the grain and

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Sorghum as a Multifunctional Crop
for the Production of Fuel Ethanol: Current Status and Future Trends 57

fiber processes require the hydrolysis of starch and fiber components into fermentable
sugars. These steps are considered expensive, take time and expend energy and other
additional resources (i.e. enzymes, chemical reagents, etc.) (Fig. 2 and 3). Despite these
benefits, some challenges must be solved in order to efficiently convert the sweet sorghum
crop into fuel ethanol. The main setbacks are the relatively higher rate of sugar degradation
at ambient temperature and the low nitrogen content for yeast growth (Mei et al., 2009; Wu
et al., 2010b). Thus, the logistics of just in time harvesting and the storage of the feedstock in
facilities that retard decomposition and degradation of fermentable carbohydrates should be
considered and stressed. In relation to nitrogen availability, this disadvantage can be
overcome with the supplementation of urea, ammonia or yeast extract in order to avoid
sluggish fermentation.
Besides sugar and nitrogen content, fermentation performance of sweet sorghum juice can
also be affected with processing parameters and bioreactor configuration. Nuanpeng et al.
(2011) observed in a repeated-batch study that very high gravity (VHG) fermentation is a
good alternative to produce high ethanol concentrations from sweet sorghum juice when an
adequate level of yeast cell concentration, nitrogen, and agitation are used. On the other
hand, Laopaiboon et al. (2007) reported better results in fed-batch fermentation compared to
batch configuration, in terms of ethanol concentration and product yield but not in
productivity (measured as grams of ethanol generated/L/hr). These findings indicate the
need to optimize parameters as feeding and withdrawn rate in order to optimize yields.

Fig. 1. Flowchart for ethanol production from sweet sorghum juice; 1Water 73%, sugars
(sucrose, glucose and fructose) 13.0%; 2Water 84%, sugars (sucrose, glucose and fructose)
14.2%. Data from: Almodares & Hadi (2009) and Gnansounou et al. (2005).
The microorganism used, as indicated in the next sections, is also a factor that is worthwhile
exploring. In the case of sweet sorghum juice, fermentation with different yeast strains has
been evaluated and productivity varies significantly, but most of the strains showed an
efficiency of more than 90% (Wu et al., 2010b). Liu et al. (2008) reported the use of
immobilized yeast in a fluidized bed reactor that shortened process time and increased
conversion efficiency. These results can be optimized when parameters as temperature,
agitation rate, particles stuffing rate and pH are modified. Liu & Shen (2008) found that
fermentation with immobilized yeast at 37°C, 200 rpm, 25% particles stuffing rate and pH of
5.0 in shaking flasks and 5 L bioreactor corresponds to the optimal conditions derived from
an orthogonal experimental design.

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58 Bioethanol

5. Ethanol fuel from sorghum grain

5.1 Conventional dry grind
The five basic steps in the conventional dry-grind ethanol process are milling, liquefaction,
saccharification, fermentation and ethanol distillation/dehydration (Fig. 2). Mashing goes
throughout the entire process beginning with mixing the grain meal with water (and possibly
backset stillage) to obtain a mash ready for fermentation. Mashing is a wet-cooking process to
turn the gelatinized starch into fermentable sugars first with the use of thermostable alpha-
amylase and then with amyloglucosidase (Zhao et al., 2008; Solomon et al., 2007; Wu et al.,
2007). Starch is the substrate for grain fuel ethanol. Unlike maize, the starch content of
sorghum is not the best indicator of ethanol yield obtained by the dry-grind process because
this carbohydrate greatly differs in availability or susceptibility to amylases.
The comparatively higher protein content of sorghum compared to maize should be
advantageous because the protein is partially degraded into free amino nitrogen compounds
during biocatalysis. These compounds are a source of nitrogen for yeast nutrition. However,
the relatively lower protein digestibility and nature of the endosperm proteins associated to
sorghum counteracts its higher protein concentration. As a result, sorghum mashes almost
always contain less free amino nitrogen compared to maize mashes. The use of proteases
during or after liquefaction is a good alternative to increase free amino nitrogen in sorghum
mashes (Perez-Carrillo & Serna-Saldivar, 2007). Protein digestibility in wet-cooked sorghum is
relatively lower compared to other cereals, mainly because of the cross-linking of prolamins.
This phenomenon reduces the availability of nitrogenous compound in sorghum mashes
needed to support yeast metabolism during fermentation.
Yeast cannot use proteins as source of nitrogen, instead it utilizes amino acids and short
peptides (di or tri), indicating the importance of protein fragmentation altogether with starch
hydrolysis in mashing. Beyond yeast nutrimental quandary, there are also issues related to
starch digestibility that affects the performance of amylolytic enzymes during liquefaction and
saccharification. This trend is also related to proteins because of the interaction between
protein and starch that in sorghum reduces the susceptibility of this polysaccharide in both
native and gelatinized conditions. Sorghum starch has higher gelatinization temperature
compared to maize and more prolamin containing bodies within the endosperm, differences
that can restrict gelatinization of starch granules (Zhao et al., 2008).
It has been reported that ethanol yields from sorghum decreases as protein content
increases; however, at the same protein level, ethanol fermentation efficiency can vary as
much as 8%. The difference is higher than typical experimental variations which indicate
that additional factors to protein affects starch-conversion rate. In a work reported by Wang
et al. (2008), nine sorghum genotypes were selected and used to study the effect of protein
availability on efficiency of ethanol fermentation. The results showed a strong positive
linear relationship between protein digestibility and fermentation efficiency, indicating the
influence, and at the same time, the usefulness of this sorghum grain features as predictor of
ethanol yield (Rooney et al., 2007; Wang et al., 2008; Wu et al., 2007; Wu et al., 2010a).
In Fig. 2 a typical process of dry-grind ethanol production is depicted. An average yield of 390
L of ethanol from 1 ton of sorghum can be obtained, but yields as high as 400 L/ton with
fermentation efficiencies of more than 90% has been achieved and reported (Chuck-Hernandez
et al., 2009; Pérez-Carrillo & Serna-Saldivar, 2007). The Dried Distillers Grains with Solubles
(DDGS) obtained in these processes contribute to the economics of biorefineries. The wet
distillers grains can be dried to 12% moisture with the aim to produce a shelf-stable byproduct.

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Sorghum as a Multifunctional Crop
for the Production of Fuel Ethanol: Current Status and Future Trends 59

Its nutritional composition (39 and 49% of protein and carbohydrates respectively) makes it an
excellent option for livestock feed, especially for ruminants.

5.2 Use of biotechnology to improve ethanol yields

5.2.1 Genetic modified sorghum
Nowadays, advances in transformation and genetic modification in plants make the
development of special sorghum cultivars one of the best tactics to overcome the various
known factors that reduce ethanol yields. Previous research works have concluded that
fermentation efficiencies and ethanol yields are influenced by genotype and chemical
composition (Wu et al., 2007, 2008; Zhao et al., 2008). These investigations have determined
important traits that enhance or reduce yields. Starch, protein and tannins are the principal
components related to ethanol production from sorghum grain and these characteristic can
be associated to genotype and also, in the case of starch and protein, to environmental
factors as sowing season and location (Wu et al., 2008). Starch composition, specifically the
amylose:amylopectin ratio, is related to fermentation efficiency. Raw materials with less
amylose are more efficiently converted into ethanol (Wu et al., 2006). The improvement is
related to digestibility of starch, reported as higher in waxy types (Rooney & Pflugfelder,
1986). Wu et al. (2006) also attributed the increased efficiency to the lower content of
amylose-lipid complexes in mashes.

Fig. 2. Flowchart for ethanol production from sorghum grain. Data from: Serna-Saldívar
(2010).
The DSC thermograms of starches from waxy sorghum and waxy maize are essentially the
same: both display a single, smooth endothermic peak, with approximately the same onset,
peak, and ending temperatures in the range of 60-80°C. However, in normal sorghum a
second peak appears around 85 to 105°C corresponding to an amylose-lipid complex that
reduces the availability of starch. Waxy starches are thereby easily gelatinized and
hydrolyzed, giving high conversion efficiencies (Wu et al., 2007). Thus, the waxy
characteristics improved the susceptibility of the endosperm matrix for low-energy
gelatinization, enzymatic hydrolysis and total ethanol production (Wu et al., 2010a).
In the case of proteins, Wu et al. (2010a) indicate that high-lysine, high-protein-digestibility
(HD) sorghum lines have been developed. These genotypes have several potential
advantages for their use as feedstocks in biorefineries. First, the starch granules swells and

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60 Bioethanol

pastes more easily at lower temperatures; second, the proteins have improved feed value
with higher bioavailability even for monogastrics. Interestingly, these high-lysine genotypes
can contain 60% more of this essential amino acid compared to regular counterparts and
similar content compared to quality protein maize (QPM) genotypes (Wu et al., 2010a). The
enhanced protein digestibility of these lines is attributed to an improved kafirin digestibility
as a result of the unique, abnormal and highly invaginated protein bodies. Segregated
progeny with HD population lack the kafirin protein body matrix that surround starch
granules and restrict swelling and pasting.
While modification in starch and protein digestibility affects ethanol production, one of the
most important traits in starch conversion is total starch harvested per area. The primary
goal of sorghum breeding programs has been and continues to be the development of high-
yielding, drought-tolerant and pest-resistant hybrids. This effort will continue and
additional gains in yield can be expected which will result in higher ethanol production
from each hectare dedicated to sorghum (Rooney et al., 2007).

5.2.2 Exogenous enzymes

As explained before, protein digestibility is related to ethanol production and this
digestibility in turn is related to the tendency of sorghum proteins to form web-like
structures during mashing which reduces the possibility of enzymes to access starch. Protein
solubility should decrease with the increase of protein cross-linking; thus, this parameter
can be used as a quality indicator in sorghum biorefineries (Zhao et al., 2008).
The utilization of proteases before conventional starch liquefaction can be used as an
alternative method to improve rate of starch hydrolysis and yield hydrolyzates with high
FAN concentration (Perez-Carrillo & Serna-Saldivar, 2007).
Perez-Carrillo et al. (2008) proposed the use of protease before starch gelatinization and
liquefaction of both decorticated and whole sorghum meals. The use of decortication to
remove the sorghum outer layers and the exogenous protease had a positive synergic effect
in terms of ethanol yield and energy savings because mashes required about half of the
fermentation time compared to conventionally processed sorghum. Decorticated meals with
more starch were more susceptible to alpha-amylase during liquefaction and produced more
ethanol during fermentation (Alvarez et al., 2010). This technology produced similar ethanol
yields compared to soft yellow dent maize and 44% more ethanol compared to the whole
sorghum control treatment. The other advantage of mechanical decortication is that the
bran, separated beforehand, is shelf-stable and can be directly channeled for production of
animal feeds and consequently the yield of wet distilled grains from decorticated sorghum
is significantly lower compared to the obtained after processing whole sorghum meals.
Thus, if dried distilled grains are produced, the biorefinery plant will spend less energy
when processing decorticated sorghum.

5.2.3 Germination and sprouting

Germinated or sprouted regular and high-tannin sorghums have improved ethanol yields
compared to the unmalted kernels. Yan et al. (2009, 2010) reported a reduction in fermentation
time and reported higher yields when sprouted sorghum was processed. The improved yield
and efficiency is attributed to the action of intrinsic enzymes in starch, proteins and cell walls.
Thus, the use of purposely malted or field sprouted sorghums can be advantageous for fuel
ethanol biorefineries. Nevertheless, the industries should consider that malting requires
important inputs in terms of water, labor, energy for drying and logistics.

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Sorghum as a Multifunctional Crop
for the Production of Fuel Ethanol: Current Status and Future Trends 61

5.2.4 Very High Gravity (VHG) fermentation

Very High Gravity (VHG) mashes are used for fuel ethanol production at industrial scale.
Among the benefits include an increased productivity, a reduced capital cost, a higher ethanol
concentration in the fermented mash (from 7-10% to 15-18% -v/v- or more), and a decrease in
water requirements. The most concentrated ethanol in fermented mashes also reduces
distillation requirements, being an important issue because after feedstock, energy is the
biggest production input, representing 30% of total ethanol cost (Pradeep et al., 2010; Wang et
al., 2007). This economic consideration indicates the importance of the substrate concentration
at the beginning of the process. The use of mashes with higher sugar concentration influences
the decision of which fermentation microorganism will be selected and used.
Yeast osmotolerance is determined by genetics and by the carbohydrate level present in
mashes, fermentation temperature, osmotic pressure/water activity and substrate
concentration. Osmotolerant yeast fermenting in batch conditions can produce and tolerate
levels of 16 to 17% (v/v) alcohol (Casey & Ingledew, 1986). According to the same authors,
higher alcohol beers can be produced if oxygenation and nitrogen sources are supplemented
to worts. Predeep et al. (2010) reported a maximum ethanol concentration of 15.6% (v/v)
converted with about 86.6% efficiency when finger millet mashes were fermented with
Saccharomyces bayanus. Fermentation temperature is also an important factor affecting
productivity, and generally speaking, at higher temperatures the time required to finish
fermentation is decreased. Jones & Ingledew (1994) reported an increment in fermentation
efficiency when dissolved solids concentration increased from 14 to 36.5 g/100 mL and also
observed that the use of urea accelerated the rate of reaction and decreased time required to
complete fermentation.
Working with VHG sweet sorghum juice rather than with ground sorghum grain, Wu et al.
(2010b) reported an increase in glycerol (0.3 to 0.6%) and residual sugars (0.2 to 5.1%) when
sugar in juices increased from 20 to 30%. A reduction in fermentation efficiency (93 to 72%)
was also observed after 72 hours fermentation. Authors recommend the use of juices with
no more than 20% soluble sugars in order to obtain the highest efficiency.
In general terms, yeasts can exhibit osmotic inhibition starting at 15% sugar, and this
inhibition is higher in glucose followed by other carbohydrates such as sucrose and maltose.
Sumari et al. (2010) stated that very few types of yeasts were known to tolerate sugar
concentration above 40% and normally at this concentration their growth is sluggish. For
this reason, the screening for osmotolerance and the development of new strains is
necessary for industrial purposes. Sumari et al. (2010), using a molecular genetic approach,
characterized a set of yeasts isolated from African brews and wines. One strain was able to
ferment a medium with sucrose concentration of 1000 g/L. The phylogenetic analysis with
rDNA clustered this microorganism away from the typical osmotolerant yeast. This
indicates the opportunity to explore and look for new strains in nature.
Besides yeast, other microorganisms, as bacteria, are especially designed for ethanol
fermentation. Escherichia coli is the typical modified microorganism for ethanol production
because of the wide spectrum of metabolized carbohydrates, its well-known genetic makeup
and the easiness of manipulation. Zymomonas mobilis, a rod shaped, gram negative, non-
spore forming bacteria is naturally ethanologenic and compared to yeast, has higher rates of
glucose uptake. Z. mobilis has also a higher ethanol production, increased yield and
tolerance, making it a good option to use in VHG fermentation. Kesava et al. (1995), working
with Z. mobilis, reported 95% conversion rates after 35 hours fermentation and ethanol
yields of approximately 70 g/L when fermenting mashes containing 150 g glucose/L. The

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62 Bioethanol

bacterium was able to ferment mashes containing 200 g glucose/L in a step-fed system.
Perez-Carrillo et al. (2011) observed that Z. mobilis had lower nitrogen requirements
compared to S. cerevisiae when fermenting mashes adjusted to 20° Plato. This bacterium has
potential and possible advantages for commercial use in biorefineries.

5.3 Physico-mechanical technologies to Improve ethanol yield

Several approaches to increase ethanol yield from sorghum involve physical or mechanical
treatments, v.gr: reduction of particle size, decortication or steam flaking. The aim of these
treatments is to reduce physical barriers to hydrolytic enzymes in order to yield more
fermentable sugars in shorter reaction times.

5.3.1 Particle size

Particle size of ground sorghum meals also plays an important role in the starch-to-ethanol
conversion process. Wang et al. (2008) observed that fermentation efficiencies of finely
ground samples were approximately 5% higher compared to coarsely ground counterparts.
This effect is a consequence of differences in gelatinization temperature and accessibility of
starch to hydrolyzing enzymes. Wang et al. (2008) reported that gelatinization temperatures
of larger or coarser particles are 5-10°C higher compared to finer particles.
The conversion of meals with smaller particles enhanced digestibility due to an
improvement in the relative surface-contact area. Mahasukhonthachat et al. (2010) indicate
that starch digestion proceeded by diffusion mechanisms is based on an inverse square
dependence of rate coefficient on average particle size.

5.3.2 Decortication
According to Rooney & Serna-Saldivar (2000) pericarp, testa, aleurone and mainly
peripheral endosperm are grain tissues directly related to the lower nutrient digestibility of
sorghum. These layers can be removed through decortication or pearling, an abrasive
process used on a regular basis for production of refined flours or grits (Serna-Saldivar,
2010). Commercial mills are typically batch type and are equipped with a set of abrasive
disks or carborundum stones to mechanically remove from 10 to 30% of the grain weight.
The resulting mixture of bran and decorticated sorghum is separated via air aspiration or
sifting (Serna-Saldivar, 2010). The classified pearled grain is then conventionally milled into
a meal or flour. This technology requires little capital investment or alteration of existing
facilities (Wang et al., 1999). The mechanical removal of the sorghum outer layers increases
starch concentration and decreases fiber, fat and phenolics. The ground decorticated
sorghum kernels are more susceptible to thermoresistant alpha-amylase hydrolysis (Perez-
Carrillo & Serna-Saldivar, 2007). Furthermore, the removal of the sorghum outer layers
allows greater starch loading and results in improved ethanol yields.

5.3.3 Steam-flaking
Other proposed alternative to process sorghum before dry-milling is steam-flaking. This
technology, widely used in feedlots, disrupts the endosperm structure with the injection of
live steam in a period of 15 to 30 min, followed by flaking through grooved rolls. Before
flaking, moisture of sorghum is increased to at least 21% and a conditioning or surfactant
agent as lecithin is added in order to obtain whole flakes and reduce processing losses
(Serna-Saldivar, 2010). After drying and cooling, sorghum flakes can be milled using

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for the Production of Fuel Ethanol: Current Status and Future Trends 63

traditional processes. The pregelatinized starch associated to the ground and steamed flaked
sorghum had higher susceptibility during liquefaction and produced more ethanol during
fermentation. Compared to the whole sorghum counterpart the steam-flaked sorghum
yielded approximately 40% more ethanol (Chuck-Hernandez et al., 2009). Currently, the cost
of steam flaking one ton of sorghum is approximately $7.5 US dollars.

5.3.4 Supercritical Fluid Extrusion (SCFX)

Extrusion has been widely used for the processing of cereal grains because this
thermoplastic technology is continuous and saves unit operations and energy. In extrusion,
the materials are subjected to heating, mixing, and shearing, resulting in physical and
chemical changes during its passage through the extruder. The major advantages of
extrusion include: improvement of starch digestibility and reduction of its molecular
weight, production of free sugars and dextrins, changes in the native structure of both starch
granules and proteins and reduced viscosity of fermentation broths. Therefore, extrusion
could be an effective process to improve the bioconversion rate of sorghum starch (Zhan et
al., 2006).
An innovative processing technology patented by researchers of Cornell University
combines extrusion process and supercritical-fluid technology. The main difference between
supercritical-fluid (SCFX) and conventional extrusion is the injection of supercritical carbon
dioxide, which replaces water as blowing agent for expansion. The injection of supercritical-
fluid carbon dioxide breaks the intimate bonds between starch granules and protein matrix
and results in the improvement of starch availability (Zhan et al., 2006). These researchers
suggested that SCFX produces molecular degradation of starch during extrusion of
sorghum. This process also increased about 8% the protein digestibility, the measurable
starch content, the free sugar concentration and gelatinized starch and other parameters that
increased ethanol yield (+5%) and boosted fermentation efficiency compared to the non-
extruded counterpart. The SCFX cooking also affected the crude fiber, chemical fraction that
after microscope examination showed disruption and fissures. These authors describe the
sorghum extrudates with “porous structure”. Thus, this thermoplastic procedure was
indeed effective as pretreatment to improve bioconversion of sorghum into ethanol.

6. Ethanol from sorghum bagasse and straw

6.1 Raw material conditioning
After extraction of juice or grain harvesting, the lignocellulosic residue is chopped, milled,
and dried at 50-60 °C to reduce the moisture content to about 10 to 15% (Herrera et al., 2003;
Sipos et al., 2009). There are many options to reduce particle size; the most commonly used
are hammer or rotary mills. Grinding can be used on both dry and wet materials, and the
cost is one of the lowest compared with others methods used for milling biomass. The
grinder reduces the particle size to a fine powder by mechanical shearing and this operation
can also be made with rotating and stationary abrasive stones (Mizuno et al., 2009).

6.2 Fiber extraction

One of the most significant problems in ethanol production from lignocellulose is
production cost (Mizuno et al., 2009) because the fiber conversion requires of high energy
investments in order to obtain high concentrations of fermentable sugars from the insoluble
polymers (Kurian et al., 2010; Mamma et al., 1996). A pre-hydrolysis step releases both the

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64 Bioethanol

hemicellulosic and cellulosic fractions of the fiber (Herrera et al., 2003). The main processes
related to the pretreatment of sorghum biomass for ethanol production are the acid and/or
enzyme-catalyzed hydrolyses (Mamma et al., 1996; Sipos et al., 2009). Generally, the acid
hydrolysis precedes the enzymatic in order to optimize production of C6 and C5
fermentable sugars (Sipos et al., 2009).

6.3 Pretreatments used for sorghum bagasse

The extraction of structural carbohydrates from bagasse cell walls is highly related to the
effectiveness of pretreatments. Nowadays there are many proposed treatments for cellulose
and hemicellulose extraction, but only few have been commercially implemented. In the
following sections some of the proposed technologies for sorghum biomass are discussed.

6.3.1 Steam explosion

The ground sorghum bagasse is rehydrated with steam at atmospheric pressure and
impregnated with low amounts (up to 3% w/w) of sulfur dioxide (SO2) in plastic bags for
20-30 minutes in order to improve enzymatic saccharification (Sipos et al., 2009; Stenberg et
al., 1998; Öhgren et al., 2005). The impregnated bagasse is introduced into a reactor and the
temperature is maintained by injection of saturated steam, varying in a range of 170-210°C
(Sipos et al., 2009; Stenberg et al., 1998; Öhgren et al., 2005). After 2 to 10 minutes, the blow-
down valve is opened and the hydrolyzate is released into a cyclone (Stenberg et al., 1998).
Sipos et al. (2009) achieved an extraction of 89% to 92% of cellulose with steam explosion, up
to 18 g glucose, 23 g xylose and 5.5 g arabinose/L hydrolyzate. Ballesteros et al. (2003) used
steam explosion pretreatment without sulfur dioxide and obtained around 50% of solids
recovery and only 20% solubilization of the cellulose. Hemicellulose sugars were extensively
solubilized because the raw material had originally 25% xylose and after the treatment only
2% remained on the fibrous residue.

6.3.2 Dilute acid hydrolysis

Acid hydrolysis, the most common fiber pretreatment method (Ban et al., 2008), generates
significant amounts of sugars from hemicellulose. Besides it is a process relatively cheap
(Gnansounou et al., 2005). Sulfuric, hydrochloric, hydrofluoric or acetic acids have been
tested as catalysts (Herrera et al., 2003). The process consists on the addition of diluted
aqueous acid solution (0.1 to 10 % w/v) to the ground raw material and hydrolyzing in an
autoclave. A solid residue, rich in cellulose and lignin, is formed after acid hydrolysis and
subsequently treated with enzymes in order to increase the amounts of fermentable sugars
(Tellez-Luis et al., 2002). Kurian et al. (2010) achieved extract with 92 g/L of total sugars
from sweet sorghum bagasse treated with sulfuric acid at a concentration of 5 g/kg and
treated at 140°C for 30 minutes. Ban et al. (2008) treated the same raw material at a solid-
liquid mass ratio of 10% with 80 g phosphoric acid/L at 120°C for 80 minutes. These authors
reported 302 g reducing sugars/kg with this pretreatment.

6.3.3 Alkali pretreatment

Unlike other pretreatments, the use of strong alkali delignifies biomass by disrupting the
ester bonds of cross-linked lignin and xylans, resulting in cellulose and hemicellulose
enriched fraction. Alkali pretreatment processes generally utilize lower temperatures,
pressures and residence times compared to other technologies (McIntosh & Vancov, 2010).

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Sorghum as a Multifunctional Crop
for the Production of Fuel Ethanol: Current Status and Future Trends 65

The main compounds used as pretreatment agents in alkali processes are: sodium
hydroxide, ammonia and lime, because of their comparatively lower cost and the possibility
of chemical and water recycling (McIntosh & Vancov, 2010). Usually two temperature
conditions are used for hydrolysis: mild (60°C) or high (121°C).

6.4 Enzymatic extraction

There are several enzymes generally used to convert cellulose and hemicellulose into
soluble sugars. They are a mixture of pectinases, cellulases and hemicellulases (Lin et al.,
2011; Reddy & Yang, 2005). Cellulose can be hydrolyzed by the synergistic action of endo-
acting enzymes knows as endoglucanases, and exo-acting enzymes, known as
exoglucanases (Lin et al., 2011). Today it is common to employ enzyme complexes
consisting of seven or more degrading enzymes that act synergistically. The enzyme mixture
is added before or after chemical or mechanical treatments (Reddy & Yang, 2005). Enzymes
appear to be the best prospects for continued improvements because can reduce production
costs (Gnansounou et al., 2005).
Sipos et al. (2009) observed that the separation of the solid and the liquid phases after
chemical pretreatment is beneficial to the whole process because the xylose-rich liquid
fraction can be fermented into ethanol through the pentose pathway or as substrate for
microbial cellulase production or transformed into other various valuable products. On the
other hand, the solid fraction can be further hydrolyzed and fermented into ethanol. The use
of alkali treatment before enzyme hydrolysis generated 540 g glucose/kg raw material,
equivalent to a 90% conversion of available cellulose to monomeric sugars. On the other
hand, 235 g xylose/kg was released after pretreatment of sorghum straw (McIntosh &
Vancov, 2010). These hydrolysates were obtained with an enzyme complex containing
endoglucanase, exoglucanase, xylanase, beta-glucosidase and cellulase.

6.5 Hydrolysis by-products or fermentation inhibitors

The fiber chemical hydrolysis process can produce a large number of sugar degradation
products which are known to inhibit bacteria and yeast and thus the conversion of
fermentable sugars into bioethanol (Ban et al., 2008). The most important inhibitors are
furfural, 5-hydroxymethylfurfural and acetic acid. After the acid hydrolysis, it is necessary
to adjust the pH with alkalis in order to obtain the adequate conditions for the subsequent
step of fermentation. Lime or calcium hydroxide is commonly added to increase the pH to 9-
10. This alkali treatment precipitates inhibitors in the form of insoluble salts and therefore
acts as detoxifying treatment (Kurian et al., 2010).

6.6 Fermentation
Hydrolyzates obtained from sorghum fiber are solutions rich in both hexoses and pentoses
(Kurian et al., 2010). Production of ethanol from these mashes is possible only with the use
of osmotolerant and pentose fermenting yeast or bacterial strains (Table 2).
Ballesteros et al. (2003) obtained 16.2 g ethanol/L when hydrolyzates obtained from sweet
sorghum bagasse were fermented with Kluyveromyces marxianus. On the other hand, Kurian et
al. (2010) working with Pichia stipitis obtained 38.7 g ethanol/L with a theoretical conversion of
82.5%. In Fig. 3, a flowchart of ethanol production from sorghum bagasse is depicted. A yield
of 158 L ethanol/ton biomass (wet basis) can be obtained after a sulfuric acid hydrolysis. The
process yielded 110 kg of lignin and other non-fermentable materials. Almodares & Hadi
(2009) and Gnansounou et al. (2005) reported that the cellulase used in Simultaneous

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66 Bioethanol

Microorganism Characteristics
Useful in fermentation of xylose to acetone and butanol; bioethanol
Clostridium acetobutilicum
produced in low yield
Capable of converting cellulose directly to ethanol and acetic acid.
Clostridium thermocellum Bioethanol concentrations are generally less than 5 g/l. Cellulase is
strong inhibition encountered by cellobiose accumulation
Native strains ferment xylose to a mixture of bioethanol, succinic, and
Escherichia coli acetic acids but lack ethanol tolerance; genetically engineered strains
predominantly produce bioethanol
Native strains rapidly ferment xylose and cellobiose; engineered to
Klebsiella oxytoca
ferment cellulose and produce bioethanol predominantly
Klebsiella planticola ATCC Carried gene from Zymomonas mobilis encoding pyruvate
33531 decarboxylase. Conjugated strain tolerated up to 4% ethanol
Consumes xylose and arabinose. Slowly uses glucose and cellobiose.
Lactobacillus pentoaceticus
Acetic acid is produced along with lactic in 1:1 ratio
Lactobacillus casei Ferments lactose, particularly useful for bioconversion of whey
Uses cellobiose if nutrients are supplied: uses glucose, D-xylose and L-
Lactobacillus xylosus
arabinose
Homolactic fermentation. Some strains produce lactic acid from sulfite
Lactobacillus pentosus
waste liquors
Consumes cellobiose more rapidly than glucose, xylose, or arabinose.
Lactobacillus plantarum Appears to depolymerize pectins; produces lactic acid from
agricultural residues
Pachysolen tannophilus
Saccharomyces cerevisiae ATCC Co-culture of S. cerevisiae and strains resulted in the best ethanol yield
24 860
NRRL strain Y-7124 utilized over 95% xylose based on 150 g/L initial
Pichia stipits NRRL Y-7124, Y-
concentration. Produced 52 g/L of ethanol with a yield of 0.39 g
11 544, Y-11 545
ethanol per g xylose
Maximum cell concentration of 50 g/L. Ethanol production rate of 10.7
Pichia stipits NRLL Y-7124
g/L.h with more than 80% xylose conversion. Ethanol and xylitol yield
(floculating strain)
of 0.4 and 0.03 g/ g xylose
Saccharomyces cerevisiae CBS
Co-culture of two yeast strains utilized both glucose and xylose. Yields
1200
of 100 and 27% on glucose and xylose, respectively
Candida shehatae ATCC 24 860
1 With data from: Balat et al. (2008) and Lee (1997).
Table 2. Native and engineered microorganisms capable of fermenting xylose to bioethanol1
Saccharification and Fermentation (SSF) can be added directly or from material previously
deviated from pretreatment and inoculated along with Trichoderma reesei or other fungi such
as Neurospora crassa and Fusarium oxysporum. These microorganisms were capable of directly
fermenting cellulose (Mamma et al., 1996). F. oxysporum was used in a SSF along with S.
cerevisiae, yielding 5.2 to 8.4 g ethanol per 100 g of fresh sorghum. The efficiency was
calculated based on soluble sugars and not in total polysaccharides (Mamma et al., 1996).

7. Estimated ethanol yields

Fig. 1 to 3 summarizes and compares average ethanol yields from sorghum grain, sweet
juice and biomass. Ethanol yields vary according to variety, geography, soil fertility and
temperature.

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Sorghum as a Multifunctional Crop
for the Production of Fuel Ethanol: Current Status and Future Trends 67

Sweet sorghums usually yield from 50 up to 120 tons of stalks after the first cut. This
feedstock contains 73% moisture, 13% soluble sugars, 5.3% cellulose, 3.7% hemicelluloses
and 2.7% lignin. The stalks yield up to 70% sweet juice and 15.33 ton/ha of spent bagasse
(Almodares & Hadi, 2009; Prasad et al., 2007).

Fig. 3. Flowchart for ethanol production from sweet sorghum bagasse. 1 Average
composition of sweet sorghum bagasse: Water 54%, simple fermentable sugars 5.4%;
Cellulose 17%; Hemicellulose 12%; Lignin 11.7% 2 Practical yield from fermentation with I.
orientalis: 3,865 L/ha. From: Almodares & Hadi (2009); Gnansounou et al. (2005).
Water added during extraction is considered part of the sweet juice yield (Fig. 1) and the
sweet juice commonly contains around 14% soluble sugars. This substrate allows the
production of 3,450 L ethanol/ha with a fermentation efficiency of 95%, similar to the result
reported Kim & Day (2011) (3,296 L/ha). These last researchers did not consider losses that
negatively affect fermentation efficiencies. Almodares & Hadi (2009), on the other hand,
reported a yield of 3,000 L ethanol/ha directly when processing juice extracted from
varieties that yielded from 39 to 128 ton stalks/ha. Although Wu et al. (2010b) did not report
ethanol yields per hectare, the calculated ethanol production from the amount of total
fermentable sugars extracted from a high yielding M81E cultivar planted at two different
locations and bioconverted with a 95% of fermentation efficiency was in the range of 4,750 to
5,220 L/ha. These potential ethanol yields are equivalent to the bioconversion of 12 to 13
tons of maize kernels.
Experimental data obtained from sweet sorghums cultivated in Central Mexico indicated
that these materials are capable of yielding 6.38 tons of sugar/ha/cut. Consequently, when
are adequately bioconverted have the potential of producing 4,132 L ethanol (unpublished
data). Regarding to the lignocellulosic fraction, if 15.33 ton of bagasse/ha is obtained
containing 29% cellulose and hemicellulose and 5.4% of remaining unextracted soluble
sugars, up to 2,400 L of ethanol can be obtained (Fig. 3). This yield represents almost half of
the 4,058 L/ha described by Kim & Day (2011) as theoretical ethanol.
In central Mexico, 42.5 ton of bagasse/ha with 50% fermentable sugars are commonly
obtained. This biomass is capable of yielding 6,375 L ethanol with perfect conversion
efficiency. However, experimental data where the acid-treated biomass was fermented with
Issatchenkia orientalis indicated only 60% fermentation efficiency (3,865 L/ha) (unpublished

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68 Bioethanol

data). These results indicate that there are still many areas for potential improvements
especially when processing spent biomass.
Almodares & Hadi (2009) reported that a yield up to 2 ton of grain/ha can be expected from
sweet sorghum. If this material is milled, hydrolyzed and fermented, a final ethanol yield of
780 L can be expected. Nevertheless, the sweet sorghum grain during optimum harvesting is
not fully matured and generally collected along the vegetative parts of the plant. Thus, the
immature sweet sorghum kernels are usually processed with the bagasse and not fermented
using grain technologies.
The biomass production per cultivated surface (Fig. 3) is the key and most important factor
that affects ethanol yields indicating the importance of both plant breeding for the generation
of new improved cultivars and the agronomic conditions mainly affected by soil fertility and
water availability. The new biomass cultivars should adapt to marginal lands in order to
minimize competition with basic grain production. The potential to obtain ethanol yields of
6630, 7000 and 10000 L/ha (with 95% of extraction and fermentation efficiency) can be
achieved because yields of 50 to 120 tons of biomass/ha are reported. Comparatively Kim &
Day (2011) indicated that the theoretical yield of maize kernels can be as high as 5,100 L/ha
and up to 8,625 L/ha when the whole plant is bioconverted into ethanol (grain + corn stover).
One of the most important factors to be addressed during yield calculation is indeed the
energy required for ethanol production. Biomass and starch require more energy for
hydrolysis compared to sweet sorghum juice. The technologies for starchy kernels and
sweet juice are matured but the conversion and estimation of energy balances when
processing lignocellulosic material will be critically important for the evaluation of
economic advisability.

8. Future trends
One of the most promising research priorities in agricultural production is the genetic
improvement of crops with high economic relevance. In the case of sorghum for fuels there
are important advances in the development of biomass, sweet and high yielding grain
varieties and hybrids, but is yet one of the most important and critical research topics. The
new cultivars should be adapted to marginal lands and also they must be resistant to pests,
other phytopathogens and stable facing water stress.
The creation of new varieties for ethanol production is not an easy task because the relevant
traits, such as plant height, total soluble solids, juice production and lignin : cellulose :
hemicellulose ratio are “non additive” (Reddy et al., 2005). On the other hand and according
to Turhollow et al. (2010), the genetic mapping combined with its relatively fast hybridation
and field tests, can facilitate the design and development of dedicated bioenergy cultivars.
It is also of upmost importance to develop machinery to harvest sweet and biomass
sorghums because the use of existing sugarcane equipments reduce yields and efficiencies.
Furthermore, it is also imperative to development new agronomical and technological
packages that include “just in time” harvesting.
The use of biomass sorghum represents one of the most relevant topics in research even
when there are not economic and energy efficient technologies. However, there have been
important advances in terms of fiber degradation to yield extracts rich in C5 and C6
fermentable sugars. The development of new and more environmental-friendly
pretreatments that include the use of fiber degrading enzymes and hot water and new
strains of yeast and bacteria are critical points for the economics of biomass transformation.

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Sorghum as a Multifunctional Crop
for the Production of Fuel Ethanol: Current Status and Future Trends 69

The new microorganisms must be designed or genetically engineered to be more efficient in

terms of enhanced capacity to fully ferment C5 and C6 sugars at high temperatures (Canizo,
2009). The development of new strains of Saccharomyces cerevisiae designed for pentose
utilization, with high tolerance to inhibitors, and with a better genomic stability has not
been yet fully addressed despite the recent advances in genetic engineering. Unfortunately,
there are only few industrial and commercial strains in the market.
Process wise, biorefineries should focus on designing new bioreactors, flow-patterns, new
cocktails of enzymes to optimize hydrolysis, the utilization of immobilized microorganisms
and the development of new distillation and ethanol dehydration technologies that favors
the total energy balance.

9. References
Almodares, A. & Hadi, M.R. (2009). Production of bioethanol from sweet sorghum: A
review, African Journal of Agricultural Research, Vol.5, No.9, (September 2009), pp.
772-780, ISSN 1991-637X
Alvarez, M., Perez-Carrillo, E. & Serna-Saldívar, S.O. (2010). Effect of decortication and
protease treatment on the kinetics of liquefaction, saccharification, and ethanol
production from sorghum, Journal of Chemical Technology and Biotechnology, Vol. 885,
No. 8, (August 2010), pp. 1122-1129, ISSN 1097-4660.
Anglani, C. (1998). Sorghum carbohydrates-A review, Plant Foods for Human Nutrition, Vol.
52, No.1, (March 1998), pp. 77-83. ISSN 0921-9668
Balat, M., Balat, H. & Öz, C. (2008). Progress in bioethanol processing, Progress in Energy and
Combustion Science, Vol.34, No. 5, (October 2008), pp.551-573, ISSN 0360-1285
Ballesteros, M., Oliva, J.M., Negro, M.J., Manzanares, P. & Ballesteros, I. (2003). Ethanol from
lignocellulosic materials by a simultaneous saccharification and fermentation
process (SFS) with Kluyveromyces marxianus CECT 10875, Process Biochemistry,
Vol.39, No. 12, (October 2004), pp. 1843-1848, ISSN 1359-5113
Ban, J., Yu, J., Zhang, X. & Tan, T. (2008). Ethanol production from sweet sorghum residual,
Frontiers of Chemical Engineering in China, Vol.2, No.4 (December 2008), pp. 452-455.
ISSN 2095-0179
Berg, C. (2004). World fuel ethanol, analysis and outlook, Japan. 28.03.11, Available from
http://www.meti.go.jp/report/downloadfiles/g30819b40j.pdf
Blake, C. (2010). Sorghum expansion tied to cellulosic ethanol. 30.06.11, Available from
http://southeastfarmpress.com/grains/sorghum-expansion-tied-cellulosic-ethanol
Canizo, J.R. (2009). Estudio del efecto de los parámetros de operación de los pretratamientos
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 Bioethanol
Edited by Prof. Marco Aurelio Pinheiro Lima

ISBN 978-953-51-0008-9
Hard cover, 290 pages
Publisher InTech
Published online 01, February, 2012
Published in print edition February, 2012

Recent studies have shown strong evidence of human activity impact on the climate of the planet. Higher
temperatures and intensification of extreme weather events such as hurricanes are among the consequences.
This scenario opens up several possibilities for what is now called "green" or low carbon economy. We are
talking about creating new businesses and industries geared to develop products and services with low
consumption of natural resources and reduced greenhouse gases emission. Within this category of business,
biofuels is a highlight and the central theme of this book. The first section presents some research results for
first generation ethanol production from starch and sugar raw materials. Chapters in the second section
present results on some efforts around the world to develop an efficient technology for producing second-
generation ethanol from different types of lignocellulosic materials. While these production technologies are
being developed, different uses for ethanol could also be studied. The chapter in the third section points to the
use of hydrogen in fuel cells, where this hydrogen could be produced from ethanol.

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Sergio O. Serna-Saldívar, Cristina Chuck-Hernández, Esther Pérez-Carrillo and Erick Heredia-Olea (2012).
Sorghum as a Multifunctional Crop for the Production of Fuel Ethanol: Current Status and Future Trends,
Bioethanol, Prof. Marco Aurelio Pinheiro Lima (Ed.), ISBN: 978-953-51-0008-9, InTech, Available from:
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ethanol-current-status-and-future-trend

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