Renewable Energy 75 (2015) 389e394

Contents lists available at ScienceDirect

Renewable Energy
journal homepage: www.elsevier.com/locate/renene

Study on the decomposition of lignocellulosic biomass and subjecting

it to alcoholic fermentation
Study on the decomposition of lignocellulosic biomass

 ska*, Wojciech Dziemianowicz
Katarzyna Kotarska, Anna Swierczy
n
Prof. Wacaw Da browski Institute of Agricultural and Food Biotechnology, Distillery Technology and Renewable Energy Division, 85-090 Bydgoszcz,
 co
w Wielkopolskich 17, Poland
Powstan

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 9 January 2014
Accepted 7 October 2014
Available online 25 October 2014

The decomposition of lignocellulosic raw material included: mechanical grinding of plant biomass,
delignication (removal of lignin e this process was conducted in alkaline environment) and detoxication process (removal of alcoholic fermentation inhibitory compounds).
The study on producing ethanol from corn straw was based on SSF method which involved conducting
simultaneous enzymatic hydrolysis of cellulose and fermentation of obtained saccharides.
Based on the study of corn straw alcoholic fermentation it was determined that the way of preparing
the raw material in the initial stage of simultaneous saccharication and fermentation, signicantly
inuences the improvement of fermentation yield.
In comparison with an attempt in which biomass detoxication process was not implemented, the
attempt with detoxication resulted in gaining higher fermentation yield and in lowering the content of
aldehydes, methanol and furfural in the produced spirit.
Moreover, in the attempts in which detoxication of raw material was used, better actual speed,
productivity and the yield of alcoholic fermentation of corn straw was noted. The conducted detoxication in the process of lignocellulosic biomass decomposition improved fermentation yield.
2014 Elsevier Ltd. All rights reserved.

Keywords:
Lignocellulosic biomass
Ethanol
Alcoholic fermentation
Biofuels

1. Introduction
In recent years, the interest in the use of renewable raw materials, including plant biomass (lignocellulosic raw materials), as an
inexhaustible source of liquid fuel (second generation biofuel) has
been constantly growing.
Moreover, second generation biofuels limit the emission of
carbon dioxide to the atmosphere, preventing the greenhouse effect. The plant biomass becomes an alternative energy source for
fossil fuels.
Whole plants, straw or agricultural product waste are very
suitable for bioethanol production, because they are not related to
food supply. They are also available in larger quantities in comparison to e.g. grain, and they are less expensive (by-products
laying on the elds, waste from plant production) [1e3].

* Corresponding author. Tel.: 48 52 341 00 82; fax: 48 52 346 00 36.

E-mail addresses: kotarska@zg.bydgoszcz.pl (K. Kotarska), swierczynska@ibprs.

 ska), dziemianowicz@zg.bydgoszcz.pl (W. Dziemianowicz).
pl (A. Swierczy
n
http://dx.doi.org/10.1016/j.renene.2014.10.018
0960-1481/ 2014 Elsevier Ltd. All rights reserved.

The lignocellulosic raw materials are built mainly out of three

connected compound groups [3e8]:
1) cellulose (comprising 30e40% of oral cell walls) e harder hydrolytic decomposition is connected to its crystalline and
amorphous areas,
2) hemicellulose (comprising 30e35% of oral cell walls) e characterized by complex carbohydrate structures (xylose, arabinose
mannose, glucose and galactose) that often create branched
chains, it is more prone to hydrolysis than cellulose,
3) lignin (comprising 11e25% of oral cell walls) e ensures plants
with impermeability and immunity against an attack from
micro-organisms and protection from chemical degradation.
The amount of specic compounds of the lignocellulose in oral
cell walls is varied due to the oral type, kind and origin. In the case
of corn stover is as follows e Table 1.
The purpose of biomass pretreatment is the degradation the
cellulosic phibrils, lowering their crystallization and polymerization levels, hemicellulose separation, degradation of cellulose

390

K. Kotarska et al. / Renewable Energy 75 (2015) 389e394

2. Materials and methods

Table 1
The amount of lignocellulose components in corn stover [9e14].
Lignocellulosic
material

Components of lignocellulose [%]

Cellulose

Hemicellulose

Lignin

Corn stover

38.90
38.14
38.70
36.40
39.00
36.80

23.20
22.68
21.70
22.60
19.10
25.40

19.10
23.34
19.30
16.60
15.10
16.90

2.1. Material

References

Tian et al., 2013 [9]

Chu et al., 2013 [10]
Zhao, Xia 2009 [11]
Lynd et al., 1999 [12]
Lee 1997 [13]
Evans et al., 1988 [14]

The study was conducted using corn straw (leaves and stems).
The material was obtained from a farm in Wojnowo. The corn straw
contained 6.33% of humidity and 93.67% of dry mass.
2.2. Enzymatic preparation
The following preparations were used for enzymatic hydrolysis
(decomposition of cellulose into glucose, cellobiose and higher
glucose polymers):

Table 2
Fermentation yield from corn straw, alkalization of the environment with the use of
Ca(OH)2, T 40  C, SSF method.
Lignocellulosic Option type
raw material

Fermentation yield
[L EtOH/100 kg of raw material]
24 h

Corn straw

48 h

72 h

10.10 0.03 13.20 0.06 14.80 0.06

without
detoxication
Detoxication
18.60 0.00 23.30 0.00 24.80 1.91
(activated carbon)

The bold values represent the fermentation yield of corn stover after 72h of
fermentation. The boldface discriminate fermentation yield of corn stover obtained
in the last day of alcoholic fermentation.

 Celluclast 1.5L (Novozymes Company, Denmark, Bagsvaerd) e

preparation obtained from Trichoderma reesei fungus cultivation, which catalyzes the decomposition of cellulose into
glucose, cellobiose and higher glucose polymers. Optimal conditions: temperature of 25e55  C and pH of 4.0e6.0.
 Novozyme 188 (Novozymes Company, Denmark, Bagsvaerd) e
enzyme obtained from Aspergillus niger fungus cultivation,
which catalyzes the decomposition of cellobiose into glucose.
This preparation is a biocatalyst that supports the effect of the
Celluclast 1.5 L enzyme. Optimal conditions: temperature of
25e55  C and pH of 4.0e5.5.

2.3. Microorganisms
complex with lignin and the modication of lignin's structure,
delignication, as well as increasing the available area for hydrolytic enzymes activity.
The pretreatment of raw materials has signicant inuence on
the efciency of the technological process [15].
After completed pretreatment, the most effective and prospective method of cellulose hydrolysis is the enzymatic method with
the help of cellulase and hemicellulose [16,17].
After the pretreatment of lignocellulosic raw material, the obtained monosaccharides are subjected to alcoholic fermentation,
using yeast.
It is very important in alcoholic fermentation process to use heat
resistant yeast breeds due to the temperature in which the enzymatic hydrolysis is conducted (optimal temperature of cellulases'
complex working is 40e50  C) [18].
The aim of the research was to conduct the decomposition of
lignocellulosic biomass in conditions appropriate for obtaining the
highest ethanol yield and benecial chemical composition of the
spirit, attesting to its quality.
When taking into account the overview of current literature,
there are many papers on various aspects of cellulose decomposition, however they do not contain information about the quality of
the obtained spirit. The spirit quality, i.e. its purity, indicates the
possibilities of its application, and the protability of its production.

The yeast strain D-2 is Saccharomyces cerevisiae, obtained from a

collection of pure cultures from IBPRS (Institute of Agricultural and
Food Biotechnology) Distillery Technology and Renewable Energy
Division in Bydgoszcz. D-2 strain is characterized by: alcohol
resistance (above 95 g/L EtOH), osmophily (22-24 Blg), thermophily (38e40  C) and acidophily (pH ok.3).

2.4. Processing of lignocellulosic raw material

The degradation of ber structure of lignocellulosic corn straw
was conducted by:
 Mechanical grinding of plant biomass into particles with
0.12 0.43 mm diameter,
 Raw material processing in alkaline environment: corn straw
(10 g) was subjected to a solution of calcium hydroxide (5 g
Ca(OH)2 130 mL of distilled H2O), and then to pressure
(0.15 MPa) and thermal (135  C) treatment in 1 h time,
 Detoxication process (removal of alcoholic fermentation
inhibiting compounds). Detoxication was performed with the
use of activated carbon (20 g/100 g of the material) in the
temperature of 80  C in 2 h time (with simultaneous shaking of
150 rpm). Then the lignocellulosic substrate was rinsed with

Table 3
Parameters describing alcoholic fermentation of corn straw e processing with the use of Ca(OH)2, with detoxication, T 40  C, SSF method.
Option

Apparent extract [ Blg]

Actual extract [ Blg]

0h

24 h

48 h

72 h

24 h

48 h

72 h

0h

24 h

Without detoxication
Detoxication
(activated carbon)

7.80 0.00
5.50 0.00

6.30 0.00
4.20 0.00

6.00 0.07
4.10 0.00

5.70 0.07
4.00 0.00

8.00 0.03
14.70 0.00

10.50 0.06
18.50 0.00

11.80 0.06
19.60 0.19

7.80 0.00
5.50 0.00

6.40 0.00
4.40 0.00

Concentration of ethanol [g/L]

Notes: alcohol concentration e the results for 0 h are missing because at that time the samples were not subjected to the fermentation process.

K. Kotarska et al. / Renewable Energy 75 (2015) 389e394

distilled water (7 L of H2O/140 mL of sample) and, after ltering,

it was dried in 105  C temperature for 1 h.
To ensure optimal conditions for the activity of used enzymatic
preparations, the plant biomass pH was set on the level of 5.0 by
using concentrated sulfuric acid.
2.5. The process of simultaneous saccharication and alcoholic
fermentation
The SSF (Simultaneous Saccharication and Fermentation)
process was performed by simultaneous hydrolysis of enzymatic
cellulose and fermentation of the obtained sugars.
The process of enzymatic hydrolysis was conducted with the use
of Celluclast 1.5L and Novozym 188 enzymes in the amount of 2 mL/
10 g of the material in the temperature of 50  C in 4 h time with
simultaneous shaking of 150 rpm.
To complete alcoholic fermentation, D-2 breed yeasts were used
in the amount of 2 gy140 mL of the sample. Alcoholic fermentation
took place in 40  C temperature, statically, in 72 h time.
2.5.1. Marking biotechnological parameters and indicators of
alcoholic fermentation process
The real extract in fermenting (in 0, 24, 48, 72 h) and attenuated
mash were measured with the aerometer (calibrated by the State
Ofce of Measures) in Balling degrees ( Blg), after ltering. The
value pH of fermenting and attenuated mash determined with the
use of Hanna Instrument digital pH meter. The reducing sugars in
slop in mashes in 0, 24, 48, 72 h of the fermentation and in the slops
was determined by Lane-Eynon's method [19]. Concentrations of
ethanol (g/L) were determined after distillation of 100 mL of ltratem mash using immersion refractometer (Carl-Zeiss) and
alcoholometric tables.
Average results of the measurements and markings were used
to calculate the biotechnological indicators of alcoholic fermentation [20], such as:

ethanol yield of cellulose [L EtOH/100 kg of cellulose],

actual speed of fermentation [L EtOH/kg of glucose  h],
fermentation productivity [L EtOH/L of mesh  h]
fermentation yield [% of theoretical].

2.6. The analysis of the quality of obtained spirit

Marking of chemical contamination in the obtained distillates was
performed with capillary gas chromatography method with the use
of Hewlett Packard 6890 gas chromatographer with EPC (Electronic
Pneumatic Control) unit, ame ionization detector (FID) and
Chrompack CP-WAX 57-CB polar capillary column (high polarity
polyethylene glycol) with the dimensions of 50 m/320 mm/0.20 mm.
Computer analytical station with Helwelt Packard Chem-Station
software was used for integrating the signal and for reporting.

391

The chromatographer is equipped with ame ionization detector that is optimized for capillary columns with EPC unit. Working
temperature of 240  C, nitrogen used as makeup gas in constant
makeup ow mode at 30 mL/min, hydrogen ow at 25 mL/min, air
ow at 280 mL/min.

3. Results
3.1. The effect of processing of the lignocellulosic raw material on
alcoholic fermentation
The study on the alcoholic fermentation of corn straw was
conducted with the use of raw material processing in alkaline
environment (calcium hydroxide), with and without the inclusion
of detoxication process. The results of fermentation yield are
shown in Table 2.
Fermentation yield obtained from corn straw with the detoxication option was higher by 40%, i.e. 24.80 L/100 kg of raw material, in comparison to fermentation yield obtained without
detoxication, i.e. 14.80 L/100 kg of raw material; (after 72 h of
fermentation), Table 2.
Similar situation can be observed when analyzing the alcoholic
fermentation process at 24th and 48th hour. During this alcohol
fermentation process at 24th hour, fermentation yield with
detoxication was higher by 45% (i.e. 18.60 L/100 kg of raw material) in comparison to the fermentation yield without detoxication
(i.e. 10.10 L/100 kg of raw material). Analogous situation was noted
at 48th hour of the process: with detoxication, the fermentation
yield was higher by 43%, in comparison to the process without
detoxication.
The analysis showed that conducting detoxication in lignocellulosic biomass decomposition process positively affected
fermentation yield.
Parameters describing alcoholic fermentation of corn straw with
the inclusion of detoxication process are shown in Table 3.
In the case of not utilizing detoxication, the apparent extract was
developing on a high level, i.e. 5.70e6.30 Blg, (24e72 h). This way, a
lower concentration of ethanol was obtained, i.e. 8.00e11.80 g/L,
(24e72 h). Produced concentration of ethanol is correlated with
yeasts not using monosaccharides available in the substrate entirely.
Its quantity (range between 14.3 g/L and 18.0 g/L, 24e72 h) left in the
biomass after completed fermentation is a testament of that, Table 3.
Implementing detoxication caused a lowering of apparent extract
by 29e33% (4.00e4.20 Blg) in correlation to the option without
detoxication (5.70e6.30 Blg), 24e72 h.
The amount of sugars directly reducing in the lignocellulose
biomass was 23.60 mg/L for 0 h process. After conducting the
fermentation process (72 h), the amount of reducing sugars
decreased by circa 82% in the case without detoxication, and by
39% in the case where detoxication was performed.
The amount of directly reducing sugars in the attenuated
biomass, with detoxication, equaled 4.20 mg/L and it was lower by

Actual extract [ Blg]

pH

48 h

72 h

0h

24 h

48 h

72 h

0h

24 h

48 h

72 h

6.20 0.00
4.30 0.00

6.00 0.00
4.10 0.07

5,00 0.00

4.80 0.00
4.70 0.00

4.70 0.07
4.70 0.00

4.70 0.00
4.60 0.00

23.60 0.00

18.00 0.06
4.90 0.00

16.20 0.04
4.40 0.00

14.30 0.08
4.20 0.01

Reducing sugars in slop [mg/L]

Notes: All biotechnological indicators of alcoholic fermentation process were determined based on the amount of alcohol obtained in the samples in specic days e alcohol concentration in 24, 48 and 72 h. The results for 0 h are
missing because at that time the samples were not subjected to the fermentation process.

35.19 0.00 44.03 0.00 46.66 0.53

0.34 0.01
0.49 0.00
0.78 0.00
4.11 2.48
5.81 0.00
9.29 0.00
33.51 2.48
31.62 0.00

72 h
48 h

27.04 0.74 35.34 1.51 39.63 1.51

0.21 0.01
0.28 0.01
0.42 0.01
2.45 1.09
3.28 1.09
5.02 0.54
28.46 1.09

48 h
72 h
48 h

25.38 1.09

Without
19.42 0.54
detoxication
Detoxication
25.27 0.00
(activated carbon)

24 h
72 h
48 h

Fermentation productivity [L EtOH/L of mash  h]

24 h
24 h

72 h

Actual speed of fermentation [L EtOH/kg of glucose  h]

24 h

In raw spirits obtained from alcoholic fermentation of corn

straw, subjected earlier to decomposition and detoxication, a gas
chromatography method was used to mark the following: higher
alcohols, acids, esters, methanol, aldehydes, acrolein and furfural.
In the above mentioned spirits, the content of carbonyl compounds
was 0.467 g/L EtOH, and it was a value lower by 79% in comparison to
the option in which the decomposition of lignocellulosic biomass was
not subjected to detoxication, i.e. 2.221 g/L EtOH, Table 5.
The calculated standard deviation in the case of aldehydes
marked in the spirits obtained with detoxication amounted to

Ethanol yield [L EtOH/100 kg of cellulose]

3.2. The evaluation of the quality of obtained raw spirit

Option

70% when compared to the option without detoxication, i.e.

14.30 mg/L (72 h).
Such low amount of directly reducing sugars noted in attenuated biomass with detoxication indicates, that the yeast use
monosaccharides available in the substrate, and attest to good
processing of alcoholic fermentation. However, in the option
without lignocellulose biomass detoxication, the reducing sugar
level indicates only a partial use of the monosaccharides available
in the substrate by the yeast.
In the option with detoxication, higher concentration of alcohol
by o 40e45% (i.e. 14.70e19.60 g/L) was noted in comparison to the
option without this process (i.e. 8.00e11.80 g/L), 24e72 h, Table 3.
The standard deviation calculated for the alcohol obtained during
3-day alcoholic fermentation without detoxication was in the range
of 0.028e0.056. In the case with detoxication, the standard deviation was calculated at 0e0.190. The occurring difference in standard
deviation for both those options is not that relevant.
During alcoholic fermentation, pH was in the range of 4.6e4.8
(24e72 h) in both cases.
The statistical analysis of the received results was done with the
use of Statistica 7.0 software. The results of calculated standard
deviation for alcoholic fermentation parameters with and without
detoxication indicate a lack of major differences between the results (i.e. 0e0.190).
It may be assumed that the use activated carbon has signicantly affected the neutralization of inhibitors occurring in biomass
subjected to cellulitic decomposition.
Biotechnological indicators of alcoholic fermentation are shown
in Table 4.
Based on the results analysis it may be observed that higher
ethanol yield by 15e23% was obtained in the option with detoxication (19.42e28.46 L EtOH/100 kg of cellulose)in comparison to
the option without one (25.27e33.51 L EtOH/100 kg of cellulose),
24e72 h, Table 3. The standard deviation shows that in the
detoxication option, a larger difference between results of
fermentation yield occurred (i.e. 2.481), compared to the option
without detoxication (i.e. 1.088), 72 h.
When comparing the speed of actual fermentation in 72nd hour
of the process, it can be observed that in the detoxication option,
this value is higher by 40% (4.11 L EtOH/kg of glucose  h) in
comparison to the value in samples without this process, i.e. 2.45 L
EtOH/kg of glucose  h.
The productivity of alcoholic fermentation obtained in the
detoxication option was higher by 38e46% (0.34e0.78 L EtOH/L of
mash  h) in comparison to the sample without detoxication
(0.21e0.42 L EtOH/L of mash  h), 24e72 h, Table 4.
With the increase of fermentation yield from corn straw, the
alcohol fermentation yield raised as well by 15% in the detoxication
option, in comparison to the option without that process (72 h).
The analysis showed that conducted detoxication in the process of lignocellulosic biomass decomposition was benecial for
ethanol yield.

Fermentation yield [% of theoretical]

K. Kotarska et al. / Renewable Energy 75 (2015) 389e394

Table 4
Biotechnological indicators of corn straw alcoholic fermentation, SSF method, with detoxication.

392

K. Kotarska et al. / Renewable Energy 75 (2015) 389e394

Table 5
The amount of by-products obtained in raw spirits during corn straw alcoholic
fermentation with the use of detoxication.
Type of chemical compound

Aldehydes
[g/L EtOH ]
Acids
[g/L EtOH ]
Esters
[g/L EtOH ]
Furfural
[g/L EtOH ]
Methanol
[g/L EtOH ]
Acrolein
[g/L EtOH ]
Higher alcohols
sum: [g/L EtOH ]
Isoamyl alcohols[g/L EtOH ]
n-propanol
[g/L EtOH ]
isobutanol
[g/L EtOH ]
n-butanol
[g/L EtOH ]

Option
Without detoxication

With detoxication

2.221 0.103

0.467 0.020

0.001 0.000

0.001 0.000

0.132 0.013

0.023 0.005

0.032 0.002

0.009 0.002

0.130 0.004

0.010 0.000

n/a

n/a

1.680 0.045

4.134 0.571

0.197 0.011
0.731 0.016

1.124 0.177
0.748 0.094

0.700 0.016

2.234 0.297

0.052 0.002

0.028 0.003

Notes: Concentration of ethanol in 72 h - variant without detoxication:

11.80 0.06 [g/L]; variant with detoxication: 19.60 0.19 [g/L].
The bold values discriminate the amount of methanol and higher alcohols obtained
in raw spirits during corn straw alcoholic fermentation. The content of these
products is important for the purpose the spirit use in the production of biofuels.

0.020 and was lower than the one from the option without
detoxication, i.e. 0.1035, Table 5.
Based on the studies on the quality of produced spirit, it was
determined that the lower value of aldehydes in it is connected to
the method of lignocellulosic structure decomposition and the
process of detoxication of the raw material that was subjected to
alcoholic fermentation.
By analyzing the chemical composition of the spirits, it may be
observed that in the detoxication option the value of furfural was
lower by 71%, i.e. 0.009 g/L EtOH in comparison to the option
without detoxication, i.e. 0.032 g/L EtOH, Table 5.
Based on those results it can be determined that the way of
preparation of the lignocellulosic raw material for alcoholic fermentation signicantly inuences the amount of furfural in the spirits.
The amount of higher alcohols produced in the tested samples of
raw spirits was low and oscillated in the range of 1.680e4.134 g/L
EtOH. The highest amount of isobutanol was noted in the general
content of fusel alcohols. The content of fusel alcohols is a crucial
parameter regarding the spirit use in the production of ethyl tertbutyl ether. Quality requirements for biofuels biocomponents
strictly determine the amount of higher alcohols, i.e., they cannot
exceed 2% [v/v].
Among the tested spirits there was no sign of acrolein presence
which attests to good sensory properties of the obtained spirits.
In the spirit obtained with the inclusion of detoxication, the
content of methyl alcohol was lower by 92% (0.010 g/L EtOH) in
comparison to the option without detoxication, i.e. 0.130 g/L EtOH,
Table 5.
As a result of using lignocellulosic biomass detoxication, the
content of chemical contamination in the obtained raw spirit was
lower than in the attempt without detoxication.

4. Discussion results and conclusions

Development work described by many authors proved that
chemical contamination from during the pre-treatment of

393

lignocellulosic biomass inhibiting effect of alcoholic fermentation

process.
In this research paper, the lignocellulose decomposition with
the use of corn straw was conducted by applying the pretreatment
of raw material, which was subjected to water steam, combined
with chemical method e using calcium hydroxide, in SSF method.
As a result, the 11.80 g/L ethanol concentration was obtained (72 h
process).
In one of the options, Jang and associates [21] described the
simultaneous saccharication and fermentation process (SSF). The
fermentation process was conducted using Saccharomyces cerevisiae yeasts, and the obtained ethanol concentration was at
0.9e1.8 g/L level.
In El-Zawawy and associates research [22] it was determined,
that the amount of obtained ethyl alcohol is dependent on the type
of pretreatment and lignocellulose material. In case of rice straw,
subjected to pulping process and the application of 10% NaOH,
7.61 g/L of ethanol was obtained, while for the material processed
with water, the ethanol concentration was 5.72 g/L.
Geng and associates [23] conducted research on obtaining
ethanol from horticultural waste. The raw material was subjected
to pretreatment (using organosolv method), enzymatic hydrolysis
(using Celluclast 1.5L and Novozym 188 enzymatic preparations)
and alcoholic fermentation using S. cerevisiae yeast. The research
led to producing 11.69 g/L ethanol. This result is close to the one
obtained in this paper.
In the above mentioned research papers of other authors
[21e23], the raw material detoxication process was not conducted. In this paper it was determined that by implementing the
detoxication of corn straw, it was possible to obtain ethanol
concentration higher by circa 40%, i.e. 19.60 g/L.
Based on the studies on lignocellulosic biomass alcoholic
fermentation, it was stated that the way of biomass decomposition
and process detoxication considerably improves ethanol yield.
The alcoholic fermentation of corn straw with the inclusion of
detoxication was characterized by higher actual speed, productivity and yield, in comparison to the option without detoxication.
Study on the decomposition of lignocellulosic biomass and the
process of detoxication (active carbon) allow to improve the
process of simultaneous saccharication and fermentation, raising
the yield of bioethanol.

References
[1] Budzianowski WM. Sustainable biogas energy in Poland: prospects and
challenges. Renew Sustain Energy Rev 2012;16(1):342e9.
[2] Sun Y, Cheng J. Hydrolysis of lignocellulosic materials for ethanol production:
a review. Bioresour Technol 2002;83(1):1e11.
[3] Tolan JS. Bioreneries e industrial processes and products. Status quo and
future direction, vol. 1; 2006. p. 193e208.
 ski L, Wo
jcik M. Production of ethanol and lignocellulosic substrates. In:
[4] Kopin
 ski B, editor. Current issues in agricultural distillery industry, farming
Czupryn
and ecology. Bydgoszcz: PM LOGO; 2002. p. 65e79.
[5] Agbor VB, Cicek N, Sparling R, Berlin A, Levin DB. Biomass pretreatment:
fundamentals toward application. Biotechnol Adv 2011;29(6):675e85.
[6] Hnedriks ATWM, Zeeman G. Pretreatments to enhance the digestibility of
lignocellulosic biomass. Bioresour Technol 2009;100(1):10e8.
[7] Xu J, Cheng JJ. Pretreatment of switchgrass for sugar production with the
combination of sodium hydroxide and lime. Bioresour Technol 2011;102(4):
3861e8.
[8] Yu Z, Jameel H, Chang H, Park S. The effect of delignication of forest biomass
on enzymatic hydrolysis. Bioresour Technol 2011;102(19):9083e9.
[9] Tian S, Li Y, Wang Z, Yang X. Evaluation of simultaneous saccharication and
ethanol fermentation of undetoxied steam-exploded corn Stover by
Saccharomyces cerevisiae Y5. Bioenergy Res 2013;6:1142e6.
[10] Chu Q, Yang D, Li X, Ma B, Yu S, Yong Q. An integrated process to enhance
ethanol production from steam-exploded corn stover. Fuel 2013;107:823e7.
[11] Zhao J, Xia L. Simultaneous saccharication and fermentation of alkalinepretreated corn stover to ethanol using a recombinant yeast strain. Fuel
Process Technol 2009;90:1193e7.

394

K. Kotarska et al. / Renewable Energy 75 (2015) 389e394

[12] Lynd LR, Wyman CE, Gerngross TU. Biocommodity Engineering. Biotechnol
Prog 1999;15:777e93.
[13] Lee J. Biological conversion of lignocellulosic biomass to ethanol. J Biotechnol
1997;56-1:1e24.
[14] Evans RJ, Knight AR, Onischak M, Babu SP. Development of biomass gasication to produce substitute fuels. Richland, Washington, USA: Pacic
Northwest Laboratory (PNL); 1988. p. 14. PNLd6518.
[15] Sybirny A, Sybirny W, Puchalski C. Biofuel ethanol from lignocellulose (plant
biomass): achievements, issues, perspectives. Prog Agric Sci 2007;4:15e23.
rska E, Mleko S, Pielecki J. The biosynthesis of protease and their
[16] Janas P, Podgo
effect on the activity of Trichoderma reesei cellulase. Ann UMCS 2004;59(1):
461e9.
[17] Zhang YHP, Lynd LR. Toward an aggregated understanding of enzymatic hydrolysis of cellulose: noncomplexed cellulose systems. Biotechnol Bioeng
2004;88(7):797e824.
[18] Ballesteros M, Oliva JM, Negro MJ, Manzanares P, Ballesteros I. Ethanol from
lignocellulosic materials by a simultaneous saccharication and fermentation

[19]

[20]
[21]

[22]

[23]

process (SFS) with Kluyveromyces marxianus CECT 10875. Process Biochem

2004;39:1843e8.
Schneider F. Sugar analysis, ofcial and tentative methods recommended by
the International Commission for Uniform Methods of Sugar Analysis
(ICUMSA). Peterborough: ICUMSA; 1979. p. 41e73.
 ski B, Kosowski G. Effect of various activators on the
Kotarska K, Czupryn
course of alcoholic fermentation. J Food Eng 2006;77(4):965e71.
Jang JS, Cho Y, Jeong GT, Kim SK. Optimization of saccharication and ethanol
production by simultaneous saccharication and fermentation (SSF) from
seaweed, Saccharina japonica. Bioprocess Biosyst Eng 2012;35:11e8.
El-Zawawy WK, Ibrahim MM, Abdel-Fattah YR, Soliman NA, Mahmoud MM.
Acid and enzyme hydrolysis to convert pretreated lignocellulosic materials
into glucose for ethanol production. Carbohydr Polym 2011;84(3):865e71.
Geng A, Xin F, Ip J. Ethanol production from horticultural waste treated by a
modied organosolv method. Bioresour Technol 2012;104:715e21.