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Bioethanol production from residual lignocellulosic materials: A review –

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The Annals of the University Dunarea de Jos of Galati
Fascicle VI – Food Technology 37(1) 25-38

REVIEW PAPER

BIOETHANOL PRODUCTION FROM RESIDUAL

LIGNOCELLULOSIC MATERIALS: A REVIEW – PART 2

CRISTIAN-TEODOR BURUIANA 1,*, GIL GARROTE 2 and CAMELIA VIZIREANU 1

1
Department of Food Science, Food Engineering and Applied Biotechnology,
Faculty of Food Science and Engineering, ‘’Dunarea de Jos’’ University of Galati,
111 Domneasca Street, 800201 Galati, Romania
2
Department of Chemical Engineering, Faculty of Science,
University of Vigo (Campus Ourense), As Lagoas, 32004 Ourense, Spain and
CITI (Centro de Investigación, Transferencia e Innovación),
University of Vigo, Tecnopole, San Cibrao das Viñas, Ourense, Spain
* Corresponding author: cristian.buruiana@ugal.ro

Received on 15th February 2013

Revised on 15 th March 2013

Lignocellulosic material (LCM) can be employed as feedstock for

biorefineries, a concept related to industries designed to process biomass for
producing chemicals, fuels and/or electrical power. According to this
philosophy, LCM can be fractionated and the resulting fractions employed for
specific applications. Bioethanol production from cellulosic fraction of LCM
involves: hydrolysis of polysaccharides and fermentation of the monomers
into bioethanol. Enzymatic hydrolysis is catalyzed by cellulolytic enzymes
and fermentation is carried out by bacteria, yeasts or fungi. The main
objective of this article is to review different process integration technologies
for bioethanol production from LCM. This paper include: separate hydrolysis
and fermentation (SHF), simultaneous saccharification and fermentation
(SSF), and simultaneous saccharification and co-fermentation (SSCF)
methods. Furthermore, the fermentation process and a comparative data of
cellulases, hemicellulases and ethanol producing-microorganisms were
presented.

Keywords: bioethanol, enzymatic hydrolysis, fermentation, separate

hydrolysis and fermentation, simultaneous saccharification and fermentation,
simultaneous saccharification and co-fermentation

Introduction
Each fuel has its intrinsic properties that play an important role in determining the
internal combustion engine performance, performance that depends on fuel quality.
An example of engine performance related to fuel quality is the relationship
between compression ratio and fuel octane. Other physical and chemical
characteristics are related to energy density, vaporization heat, molecular ratio
26 C. T Buruiană et al. / AUDJG – Food Technology 37(1) 25-38

between reactants and combustion products, specific energy, flammability limits,

flame transmission speed, flame temperature and the hydrogen and carbon content.
The use of bioethanol mixed with gasoline has a number of advantages: reduction
by 8-30% of carbon monoxide emissions when using 10% ethanol in gasoline;
reduction by 5-15% of toxic emissions (sulfur and aromatic compounds such as
benzene, toluene, xylene) without influencing fuel performance; reduction of the
ozone layer; reduction of pollution. Fuel bioethanol is currently used worldwide in
E 10 mixture (10% anhydrous ethanol and 90% gasoline). This mixture (E 10)
contains 3.5% oxygen compared to 2.7% for gasoline. Bioethanol can be mixed
with gasoline in higher proportions: 85% (E 85), 95% (E 95) and used entirely (E
100) as an alternative fuel source (Banu et al., 2006).
After pretreatment, the next steps in the biochemical process of bioethanol
production from lignocellulosic materials (LCM) are: enzymatic hydrolysis of
polysaccharides and fermentation of monosaccharides into bioethanol. They can be
performed separately or simultaneously (Tomás-Pejó et al., 2008). Enzymatic
hydrolysis can be applied at different levels of process integration: separate
hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation
(SSF), simultaneous saccharification and co-fermentation (SSCF) (Hamelinck et
al., 2005). Enzymatic hydrolysis is catalyzed by cellulolytic enzymes and the
fermentation is carried out by yeast or bacteria. Cellulases are composed of a
mixture of enzyme proteins responsible for the degradation of cellulose to glucose
(Howard et al., 2003). The efficiency of enzymatic hydrolysis depends on the
appropriate proportional ratio of the cellulose components. SHF process is
performed in two different vessels and each step can be done under optimal
conditions of pH and temperature. In the SSF, the enzymatic saccharification and
fermentation process are run in the same vessel and glucose released by the action
of cellulases is converted directly into ethanol by the fermenting microorganisms.
These advantages result in an increased rate of saccharification and ethanol
productivity compared to SHF (Wyman, 1996). SSCF process combines the
hydrolysis of cellulose to glucose and the co-fermentation of pentose and hexose
sugars to ethanol in one reaction vessel (Lynd, 1996; Spatari et al., 2010). The
main advantages of these processes are: in SHF each stage can be processed at its
optimal operating conditions and separate steps minimize interaction between the
hydrolysis and fermentation, compared to SSF where can be obtained higher
ethanol yields due to removal of end product inhibition of saccharification process
and can be reduced the number of reactors required (Sarkar et al., 2012).
This review is focused on bioethanol production from cellulosic fraction of
lignocellulosic materials (LCM) based on hydrolysis of polysaccharides (usually
using enzymes) and fermentation of the monomers into bioethanol.

Enzymatic hydrolysis
Cellulases and cellulase-producing microorganisms
Cellulases represent an enzymatic complex that is capable of hydrolyzing cellulose.
This enzyme complex includes:
 C. T Buruiană et al. / AUDJG – Food Technology 37(1) 25-38 27

 Endo-1,4-β-glucanases (EG), acting within polyglucidic chains forming

cellulose, making the final breaking of β-1,4-glucosidic bonds and
formation of shorter chains with free reducing ends;
 Exo-1,4-β-D-glucanases or cellobiohydrolases (CBH), which include two
enzymes: 1,4-β-D-glucanglucohydrolase which releases glucose units from
the reducing end of cellulose chains, but slowly hydrolyses cellobiose
formed by another enzyme; and 1,4-β-D-glucancellobiohydrolase which
releases cellobiose units from the reducing end of cellulase chains;
 β-D-glucosidases (BGL) and β-D-glucosideglucohydrolase (cellobiase)
that hydrolyze cellobiose and short chains of oligosaccharides to glucose
(Zhang & Lynd, 2004; Howard et al., 2003; Heikinheimo, 2002; Gusakov
et al., 2007; Gusakov et al., 2005; Valjamae et al., 2001).
When the enzymatic system (cellulase) acts in vitro on insoluble cellulose
substrate, three significant processes occur simultaneously: chemical and physical
changes in the cellulosic fraction; primary hydrolysis, which involves the release of
soluble sugars from the surface of cellulosic molecules; secondary hydrolysis,
which involves hydrolysis of soluble sugars to lower molecular weight sugars, and
finally to glucose (Mosier et al., 2002).
Several species of bacteria such as Clostridium, Cellulomonas, Thermonospora,
Bacillus, Bacteriodes, Ruminococcus, Erwinia, Acetovibrio, Microbispora,
Streptomyces species, and fungi such as Trichoderma, Penicillium, Fusarium,
Humicola, Phanerochaete, Schizophillum species are able to produce cellulases
(Rabinovich et al., 2002; Sun & Cheng, 2002). Cellulases can also be produced by
aerobic bacteria such as Pseudomonas and Actinomycetes species as well as fungi
of Aspergillus species (Sun, 2002).

Hemicellulases and hemicellulase-producing microorganisms

Hemicellulases can be classified into three categories:
 Endo-hemicellulases which act within the hemicellulose chain and have
limited activity on short chain oligomers;
 Exo-hemicellulases which act progressively outside the hemicellulose
chain;
 Hemicellulases that hydrolyze hemicellulose from native plants (acetyl
esterases and esterases).
Several species of bacteria (Table 1) such as Agrobacterium, Bacillus,
Bifidobacterium, Clostridium, Streptomyces, Cellulomonas, Geobacillus species,
and fungi such as Aspergillus and Trichoderma species are able to produce
hemicellulases (Shallom & Shoham, 2003).
The main hemicellulase-producing microorganisms from several species of fungi
are as follows: Aspergillus niger, Trichoderma reesei and Trichoderma viride
species (Banu et al., 2006).
 28 C. T Buruiană et al. / AUDJG – Food Technology 37(1) 25-38

Table 1. Comparative data of different microorganisms with the highest specific activity
for cellulases and hemicellulases (source: Howard et al., 2003; Menon, & Rao, 2012)

Specific activity
Organism Enzyme Substrate
(μmol.min-1.mg-1)
Cellulase-producing microorganisms
Aspergillus niger Cellulase Carboxymethylcellulose / cellohexaose / cellopentaose 194
/
Cellotetraose / cellotriose / cellulose
Achlya bisexualis 1,3-β-glucan Glucan / laminarin / neutral glucan / phosphoglucan 7840
glucohydrolase
Rhizopus chinensis 1,3-β-D-glucan β-glucan 4800
glucanohydrolase
Penicillium 1,6-β-D-glucan β-glucan/gentiobiose / pachyman 405
brefeldianum glucanohydrolase
Bacillus subtilis Mannan endo- Galactoglucomannan / glucomannans / mannans 514
1,4-β-
mannosidase
Clostridium Cellulase Avicel / carboxymethylcellulose / cellulose / 428
thermocellum cellopentaose /
Cellotetraose / cellotriose
Streptomyces 1,3-β-glucan Laminarin 6.7
murinus glucohydrolase
Bacillus macerans 1,3-1,4-β-D- β-D-glucan/lichenan 5030
glucan
glucanohydrolase
Hemicellulases-producing microorganisms
Trichoderma Endo-1,4-β- 1,4-β-D-xylan 6630
longibrachiatum xylanase
Aspergillus nidulans β-1,4-xylosidase p-nitrophenyl-β-D-xylopyranoside 107.1
Aspergillus niger Exo-β-1,4- β-D-Man-(1-4)-β-D-GlcNAc-(1-4)-β-DGlcNAc-Asn- 188
mannosidase Lys
Aspergillus niger Endo-α-1,5- 1,5-α-L-arabinan 90.2
arabinanase
Sclerotium rolfsii Endo-β-1,4- Galactoglucomannan / mannans / galactomannans / 380
mannanase glucomannans
Phanerochaete α-Glucuronidase 4-O-methyl-glucuronosyl-xylobiose 4.5
chrysosporium
Mortierella vinacea α-Galactosidase Melibiose 2000
Humicola insolvens β-glucosidase (2-hydroxymethylphenyl)-β-D-glucopyranoside 266.9
Schizophyllum Acetyl xylan 4-methylumbelliferyl acetate / 4-nitrophenyl acetate 227
commune esterase
Clostridium Feruloyl esterase Ethyl ferulate 88
stercorarium
Thermoanaerobacter β-1,4-xylosidase o-nitrophenyl-β-D-xylopyranoside 1073
ethanolicus
Fibrobacter Acetyl xylan Acetylxylan / α-naphthyl acetate 2933
succinogenes esterase
Pyrococcus furiosus Exo-β-1,4- p-nitrophenyl-β-D-galactoside 31.1
mannosidase
Bacillus subtilis Endo-β-1,4- Galactoglucomannan / glucomannans / mannan 514
mannanase
Bacillus subtilis Endo-α-1,5- 1,5-α-L-arabinan 429
arabinanase
Bacillus subtilis Endo-galactanase Arabinogalactan 1790
Bacillus polymyxa β-Glucosidase 4-nitrophenyl-β-D-glucopyranoside 2417
Bacillus pumilus Endo-1,4-β- β-1,4-D-xylan 1780
xylanase
Escherichia coli α-Galactosidase Raffinose 27350
 C. T Buruiană et al. / AUDJG – Food Technology 37(1) 25-38 29

Fermentation
Hydrolysate obtained by acid pretreatment is used for fermentation by
microorganisms. Because the hydrolysate contains not only glucose, but also
different monosaccharides such as xylose, galactose, mannose, arabinose and
oligosaccharides, microorganisms are necessary to ferment these sugars (Katahira
et al., 2006). These microorganisms can use carbohydrates with 6-carbon atoms,
one of the most common being glucose. Cellulosic materials containing high levels
of glucose or glucose precursors are most easily converted into bioethanol (Balat et
al., 2008). There is a number of microorganisms that produce significant amounts
of bioethanol (Steward & Russell, 1987). Xylose fermenting microorganisms are
bacteria, yeasts and filamentous fungi (Hahn-Hagerdal et al., 2006). One of the
most efficient ethanol producing yeast is Saccharomyces cerevisiae which has a
high tolerance to ethanol and other inhibitory compounds resulting from acid
hydrolysis of LCM. Since wild strains of this yeast cannot ferment pentose such as
xylose, arabinose and oligosaccharides, production of bioethanol from
lignocellulosic hydrolysate is inadequate (Katahira et al., 2006).
Ethanologenic bacteria which are currently used for bioethanol production from
LCM are: Escherichia coli, Klebsiella oxytoca and Zymomonas mobilis (Dien et
al., 2003). Zymomonas mobilis is well known for its ability to quickly and
efficiently produce ethanol from raw materials based on glucose and comparative
tests have shown that Zymomonas mobilis can achieve yields higher than 5% and a
productivity up to five times higher compared to traditional yeast fermentation.
Zymomonas mobilis showed ethanol yields up to 97% and ethanol concentrations
up to 12% for glucose fermentation (Mohagheghi et al., 2002). Zymomonas mobilis
can efficiently produce ethanol from hexose: glucose and fructose, but not from
pentose (Hahn-Hagerdal et al., 2006). Zymomonas mobilis ferments only glucose,
fructose and sucrose. In the recent years, researchers at NREL have successfully
designed Zymomonas mobilis strains capable of fermenting xylose and arabinose
(Dien et al., 2003). Escherichia coli and Klebsiella oxytoca metabolize arabinose
so that ethanologenic strains ferment all lignocellulose sugars (Hahn-Hagerdal et
al., 2006). Pichia stipitis, Candida parapsilosis and Candida shehatae can
metabolize xylose by the action of xylose reductase to convert xylose to xylitol and
by the action of xylitol dehydrogenase to convert xylitol to xylulose. Xylose
fermentation can be carried out successfully by recombinant Saccharomyces
cerevisiae which takes xylose reductase and xylitol-dehydrogenase from Pichia
stipitis, and xylulokinase from Sacharomyces cerevisiae (Katahira et al., 2006).

Table 2. Ethanol-producing microorganisms for the bioconversion of LCM

(source: Waites et al., 2001)

Microorganism Fermented substrate

Bacteria
Clostridium thermohydrosulfuricum Glucose, xylose sucrose and cellobiose
(thermophilic)
Clostridium thermocellum Glucose and cellobiose
(thermophilic)
30 C. T Buruiană et al. / AUDJG – Food Technology 37(1) 25-38

Microorganism Fermented substrate

Thermoanaerobacter ethanolicus Xylose
Thermoanaerobium brockii Glucose, sucrose and cellobiose
(thermophilic)
Thermobacteroides aceroethylicus Glucose, sucrose and cellobiose
(thermophilic)
Zymomonas mobilis Glucose, fructose and sucrose
Yeasts
Candida pseudotropicalis Glucose and galactose
Candida tropicalis Glucose, xylose and xylulose
Kluyveromyces fragilis Glucose and galactose
Pachysolen tannophilus Xylose*
Pichia stipitis Xylose*
Saccharomyces cerevisiae Glucose, sucrose, galactose, fructose, xylulose,
maltose and maltotriose
Saccharomyces caerlbergensis Glucose, galactose, fructose, xylulose, maltose
and maltotriose
Saccharomyces rouxii (osmophilic) Glucose, sucrose, fructose and maltose
Filamentous fungi
Fusarium sp. Xylose*
Mucor sp. Xylose and arabinose*
* In addition to hexose monosaccharides and disaccharides

Process integration
Separate hydrolysis and fermentation (SHF)
Enzymatic hydrolysis of cellulose consists in a heterogeneous reaction where
endoglucanases randomly attack the inner links of the chain and exoglucanases
catalyze the oligosaccharides from the end of the polymer chains releasing
cellobiose and shorter chains of cellulose which are soluble or do not depend on its
degree of polymerization. Cellobiose is passing in the aqueous solution and is
hydrolyzed in the homogeneous phase by β-1,4-glucosidase which catalyzes the
hydrolysis of cellobiose to glucose. Most investigated cellulase systems were
extracted from fungi such as Trichoderma viride, Trichoderma reesei and
Fusarium solani (Gan et al., 2003). Enzymatic hydrolysis is carried out under
conditions of moderate temperature and pH in non-corrosive medium (temperature
= 45-50°C and pH = 4.8) (Duff & Murray, 1996). This represents significant
savings in energy and equipment. Furthermore, the enzymatic hydrolysis will only
attack polysaccharides without altering the phenolic fraction, resulting cleaner and
easily fermentable solutions (Vazquez et al., 1991). In the first step glucose
solution enters in fermentation reactor and the mixture is then distilled to remove
bioethanol. In the second step, xylose is fermented with the production of
bioethanol and then the bioethanol is distilled (Hamelinck et al., 2003; Hamelinck
et al., 2005). The main advantage of SHF is that hydrolysis and fermentation take
place under optimum conditions. The disadvantage is that cellulolytic enzymes are
inhibited, so that the rate of hydrolysis is reduced when glucose and cellobiose are
accumulating (Hahn-Hagerdal et al., 2006; Balat et al., 2008) (Figure 1).
 C. T Buruiană et al. / AUDJG – Food Technology 37(1) 25-38 31

Figure 1. Separate hydrolysis and fermentation (SHF) with separate pentose and hexose
sugars and combined sugar fermentation (source: Chandel et al., 2007; Balat et al., 2008)

Simultaneous saccharification and fermentation (SSF)

Enzymatic hydrolysis of LCM is a very slow process because cellulose hydrolysis
is prevented by the structural parameters of the substrate, such as lignin and
hemicellulose content as well as by crystallinity of cellulose (Pan et al., 2006).
Enzymatic hydrolysis usually occurs at pH = 4.8 and temperature of 45-50˚C (Sun,
2002). Carbohydrates of pretreatment and enzymatic hydrolysis steps are
fermented by enzymes produced by certain microorganisms (bacteria, yeasts or
filamentous fungi) (Hahn-Hagerdal et al., 2006). SSF provides higher bioethanol
yields and requires small amounts of enzyme (Dien et al., 2003; Chandel et al.,
2007). SSF process can be carried out in batch mode with multiple additions of
solid substrate or in fed-batch mode with higher concentrations of final product.
The optimum enzyme loading depends on the conditions during SSF such as water-
insoluble solids (WIS) and inhibitory compounds concentration in the SSF medium
(Hoyer et al., 2010).
SSF is a process that uses natural materials containing heterogeneous polymer
complex, such as lignin, pectin and lignocellulose (Sabu et al., 2006) (Figure 2).
Major advantages of SSF are (Sun & Cheng, 2002): increase of hydrolysis rate by
conversion of sugars; small amounts of enzymes used; high yields of the products
obtained; lower requirements under sterile conditions because glucose is removed
immediately by producing bioethanol; short processing time; lower volume
bioreactor. The main disadvantage of SSF process is to choose the optimal
temperature (Krishna et al., 2001). In many cases, pH < 5 and temperatures higher
than 40˚C may be favorable for enzymatic hydrolysis, since low pH can inhibit the
production of lactic acid and high temperatures can adversely affect the growth of
microorganisms (Huang et al., 2005). Cellulase from Trichoderma reesei, which is
the most active enzyme preparation, has optimal activity at pH = 4.5 and
32 C. T Buruiană et al. / AUDJG – Food Technology 37(1) 25-38

temperature of 55˚C. For Saccharomyces cerevisiae cultures, the process is

controlled at pH = 4.5 and temperature of 37˚C (Dien et al., 2003).

Figure 2. Simultaneous saccharification and fermentation (SSF) with combined sugars

fermentation (source: Chandel et al., 2007; Balat et al., 2008)

Several species of bacteria such as Clostridium, Thermoanaerobacter,

Thermoanaerobium, Thermobacteroides species, yeasts such as Candida,
Kluyveromyces, Pachysolen, Pichia, Saccharomyces species and fungi such as
Fusarium and Mucor species are able to produce ethanol (Waites et al., 2001).

Simultaneous saccharification and co-fermentation (SSCF)

Recently, SSF technology has proven to be advantageous for simultaneous
fermentation of hexose and pentose sugars, this process is called simultaneous
saccharification and co-fermentation (SSCF). In SSCF, enzymatic hydrolysis is
releasing continuously hexose sugars, which increase the rate of glycolysis, so that
pentose sugars are fermented faster and with better yields (Hahn-Hagerdal et al.,
2006). In this process, cellulose is converted into glucose using enzymes
(cellulase). An enzyme preparation (cellulase) is a mixture of enzymes (catalytic
proteins) that work together to break the cellulose fibers in cellobiose and glucose.
Glucose and carbohydrates resulted in hydrolysis of hemicellulose during
pretreatment are fermented to bioethanol. Bacteria used for co-fermentation of
cellulose are Zymomonas mobilis because this organism and its genome is largely
accessible (Urbanchuk, 2006). "Co-fermentation" means that the microorganism
can ferment simultaneously glucose and xylose to bioethanol. For fermentation of
cellulose are considered other cultures such as genetically modified strains of
Saccharomyces cerevisiae (McAloon et al., 2000; Balat et al., 2008).
Enzymatic hydrolysis is initiated in a continuous reactor. Diluted and neutralized
hydrolysate is cooled with water and introduced into the reactor at a concentration
of 20%. Originally, the enzyme (cellulase) is mixed with the hydrolysate at a
temperature of 48°C (Humbird et al., 2011). The quantity of enzyme used is
 C. T Buruiană et al. / AUDJG – Food Technology 37(1) 25-38 33

determined by the quantity of cellulose present in the hydrolysate and the specific
activity of the enzyme. During enzymatic hydrolysis, temperature is maintained by
cooling water. Studies by NREL showed optimum temperature at 48 °C for a range
of commercial enzymes. A deviation from this temperature would reduce the
efficiency of converting cellulose into bioethanol. The hydrolysate is containing
11.7% total soluble carbohydrates (including oligomers) with 6.7% glucose and
3.7% xylose. The hydrolysate is cooled to a temperature of 32°C for fermentation
(Humbird et al., 2011). Recombinant bacteria Zymomonas mobilis can
simultaneously ferment glucose and xylose to bioethanol. In order to ensure an
appropriate volume of 10% inoculum for fermentation, 10% of hydrolysate is sent
to prepare the culture medium. In addition to being fermented to ethanol, sugars
can be converted into products of contamination by microorganisms. Co-
fermentation process lasts 36 hours. Inoculum is fed with corn syrup (CSL) and
ammonium phosphate (DAP). It is assumed that the viscosity reduction of
hydrolysate occurs in continuous hydrolysis reactor and stirring is not required in
anaerobic fermentation with Zymomonas mobilis. Fermented syrup, having a 5.4%
ethanol concentration, is sent to the storage tank (Humbird et al., 2011).
Olofsson et al. (2010a) demonstrates a new approach for controlling the glucose
release rate from the enzymatic hydrolysis by controlling the addition of enzymes
in SSCF using spruce as feedstock and a recombinant xylose fermenting strain
Saccharomyces cerevisiae TMB3400. The results showed that the total xylose
uptake could be increased from 40% to 80% by controlling the enzyme feed.
Jin et al. (2012) presented information on the performance of industrial xylose
fermenting strain in lignocellulosic hydrolyzates. Xylose consumption by
Saccharomyces cerevisiae 424A(LNH-ST), which is a widely used genetically
modified xylose-fermenting yeast strain for ethanol production from AFEXTM
pretreated biomass, during SSCF of AFEXTM pretreated switchgrass was inhibited
by unhydrolyzed solids. Such inhibitory effects were not found in unhydrolyzed
solids from AFEXTM pretreated corn stover.

Table 3. Comparative data of ethanol production from LCM using different process
configurations

Final
Ethanol Ethanol
Temp ethanol
Substrate Microorganism Process yield yield References
(°C) conc
(g g-1)1 (%)2
(g L-1)
Sugarcane Zymomonas SHF EH:45; 6.24 0.403 79.09 Wirawan et
bagasse mobilis F:30 al., 2012
(immobilized in
PVA)
Sugarcane Zymomonas SHF EH:45; 5.52 0.356 69.96 Wirawan et
bagasse mobilis F:30 al., 2012
(immobilized in
CA)
Sugarcane Zymomonas SSF 30 5.53 0.357 70.09 Wirawan et
bagasse mobilis al., 2012
(immobilized in
PVA)
34 C. T Buruiană et al. / AUDJG – Food Technology 37(1) 25-38

Final
Ethanol Ethanol
Temp ethanol
Substrate Microorganism Process yield yield References
(°C) conc
(g g-1)1 (%)2
(g L-1)
Sugarcane Zymomonas SSF 30 5.44 0.351 68.95 Wirawan et
bagasse mobilis al., 2012
(immobilized in
CA)
Bermudag Saccharomyces SSF 38 16.10 0.480* 94.70 Li et al.,
rass cerevisiae 2009
Reed Saccharomyces SSF 38 16.40 0.490* 96.40 Li et al.,
cerevisiae 2009
Rapeseed Saccharomyces SSF 38 15.80 0.470* 92.90 Li et al.,
cerevisiae 2009
Rice straw Saccharomyces SSF 38 12.70 0.420* 83.10 Ko et al.,
cerevisiae 2009
Corn Saccharomyces SSF 35 33.80 0.410* 80.20 Öhgren et
stover cerevisiae al., 2007
Barley Saccharomyces SSF 35 22.40 0.410* 80.00 Linde et al.,
straw cerevisiae 2007
Spruce Saccharomyces SSF 37 44.50 0.430 84.00 Rudolf et
cerevisiae al., 2005
Populus Kluyveromyces SSF 42 19.00 0.360 71.20 Ballesteros
nigra marxianus CECT et al., 2004
10875
Eucalyptu Kluyveromyces SSF 42 17.00 0.320 62.50 Ballesteros
s globulus marxianus CECT et al., 2004
10875
Wheat Kluyveromyces SSF 42 18.10 0.320 62.50 Ballesteros
straw marxianus CECT et al., 2004
10875
Sweet Kluyveromyces SSF 42 16.20 0.310 60.90 Ballesteros
sorghum marxianus CECT et al., 2004
bagasse 10875
Brassica Kluyveromyces SSF 42 19.00 0.350 68.10 Ballesteros
carinata marxianus CECT et al., 2004
residue 10875
Spruce Saccharomyces SSCF 34 32.90 0.390 77.00 Olofsson et
cerevisiae al., 2010a
TMB3400
Spruce Saccharomyces SSCF 34 35.50 0.410 79.00 Olofsson et
cerevisiae al., 2010a
TMB3400
Wheat Saccharomyces SSCF 34 38.00 0.350 69.00 Olofsson et
straw cerevisiae al., 2010b
TMB3400
* Not directly given in the reference article, calculated by the authors.
1
g ethanol/g cellulose (in most cases expressed as potential glucose) in the pretreated raw material.
2
based on maximum theoretical ethanol yield on available sugars (in most cases only glucose) in the pretreated
raw material.
PVA – polyvinyl alcohol; CA – calcium alginate; EH – enzymatic hydrolysis; F – fermentation.

Conclusions
The challenges for different process integration technologies for bioethanol production
from LCM are to obtain high degree of hydrolysis and high ethanol yields.
Compared with SHF technology, where each stage takes place under optimal operating
conditions (minimizing the interaction between hydrolysis and fermentation), the main
 C. T Buruiană et al. / AUDJG – Food Technology 37(1) 25-38 35

SSF advantages are as follows: i) obtaining higher ethanol yields with small amount of
enzymes; ii) increasing the hydrolysis rate by sugars conversion; and iii) lower
requirements under sterile conditions, because glucose is removed immediately by
producing bioethanol. In SSCF process, enzymatic hydrolysis is releasing continuously
hexose sugars, so that pentose sugars are fermented faster and with better yields.

Acknowledgments
The work of Cristian-Teodor Buruiana was supported by Project SOP HRD - TOP
ACADEMIC 76822/2010.

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