Bioresource Technology 100 (2009) 1007–1010

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Short Communication

Ethanol production from cotton-based waste textiles

Azam Jeihanipour a,b,*, Mohammad J. Taherzadeh a
a
School of Engineering, University of Borås, Borås, Sweden
b
Chemical Reaction Engineering, Chalmers University of Technology, Göteborg, Sweden

a r t i c l e i n f o a b s t r a c t

Article history: Ethanol production from cotton linter and waste of blue jeans textiles was investigated. In the best case,
Received 13 May 2008 alkali pretreatment followed by enzymatic hydrolysis resulted in almost complete conversion of the cot-
Received in revised form 9 July 2008 ton and jeans to glucose, which was then fermented by Saccharomyces cerevisiae to ethanol. If no pretreat-
Accepted 12 July 2008
ment applied, hydrolyses of the textiles by cellulase and b-glucosidase for 24 h followed by simultaneous
Available online 23 August 2008
saccharification and fermentation (SSF) in 4 days, resulted in 0.140–0.145 g ethanol/g textiles, which was
25–26% of the corresponding theoretical yield. A pretreatment with concentrated phosphoric acid prior to
Keywords:
the hydrolysis improved ethanol production from the textiles up to 66% of the theoretical yield. However,
Cotton linter
Jeans waste textiles
the best results obtained from alkali pretreatment of the materials by NaOH. The alkaline pretreatment of
Enzymatic hydrolysis cotton fibers were carried out with 0–20% NaOH at 0 °C, 23 °C and 100 °C, followed by enzymatic hydro-
Ethanol lysis up to 4 days. In general, higher concentration of NaOH resulted in a better yield of the hydrolysis,
Alkaline pretreatment whereas temperature had a reverse effect and better results were obtained at lower temperature. The
best conditions for the alkali pretreatment of the cotton were obtained in this study at 12% NaOH and
0 °C and 3 h. In this condition, the materials with 3% solid content were enzymatically hydrolyzed at
85.1% of the theoretical yield in 24 h and 99.1% in 4 days. The alkali pretreatment of the waste textiles
at these conditions and subsequent SSF resulted in 0.48 g ethanol/g pretreated textiles used.
Ó 2008 Elsevier Ltd. All rights reserved.

1. Introduction
Used clothes and surplus of textile industries end sooner or la-
ter in waste collection stations, and usually landfilled or inciner-
ated (Miranda et al., 2007). Cotton with annual production of
more than 23 million tons per year (FAO, 2003) stands for about
1/3 of the global market of textile fibers (Aizenshtein, 2004). Cot-
ton fibers are the seed hairs of the plant Gossypium. Cotton is typ-
ically composed of 88–96% cellulose, the remainder being protein,
pectin materials and wax. Cotton must be scoured and usually
bleached before use, whereby its cellulose content is enriched to
about 99% (Miranda et al., 2007). It is therefore possible to hydro-
lyze the cotton by e.g. enzyme or acids to glucose and then ferment
it to ethanol.
Ethanol is currently the most abundant renewable in the global
fuel market. It is currently produced from sugars and starchy mate-
rials, but lignocelluloses are most likely the alternative feedstocks
for the second generation of ethanol production in the future (Lynd
et al., 2005; Taherzadeh and Karimi, 2007a). The polymers of cellu-
lose and hemicelluloses should be first released from the fibrils in
‘‘pretreatment” in order to have an effective hydrolysis (Taher-
* Corresponding author. Address: School of Engineering, University of Borås,
Borås, Sweden. Tel.: +46 33 435 4636; fax: +46 33 435 4008.
E-mail address: Azam.Jeihanipour@hb.se (A. Jeihanipour).
zadeh and Karimi, 2007b). The general ideas in various pretreat-
ment technologies are to alter or remove lignin and
hemicelluloses, increasing the surface area and decreasing the
crystallinity of cellulose (Galbe and Zacchi, 2002; Jørgensen et al.,
2007; Wyman et al., 2005). In cotton, there is no lignin or hemicel-
luloses, but its crystallinity is relatively high. The crystallinity
index of Avicel, wood pulp, bacterial cellulose and cotton is 0.50–
0.60, 0.50–0.70, 0.76–0.95 and 0.81–0.95, respectively (Zhang
and Lynd, 2004). Therefore, the main goal of any possible pretreat-
ment of cotton-based textiles should be to increase the surface
area and decrease the crystallinity. In order to approach this goal,
one should efficiently disrupt hydrogen bonds between the glucan
chains in crystalline cellulose, and produce amorphous cellulose.
Zhang et al. (2006, 2007) developed a procedure for producing
regenerated amorphous cellulose, by dissolving homogenous cellu-
lose in concentrated phosphoric acid followed by a precipitation by
acetone. They concluded that in this method, by breaking down the
hydrogen bonds and precipitating amorphous cellulose, it is possi-
ble to approach complete enzymatic hydrolysis. This pretreatment
method was used in our work on waste cotton textiles. Another
concept that we applied was using sodium hydroxide as a swelling
agent for cotton, which is known in textile industries (Porro et al.,
2007; Roy et al., 2003; Warwicker, 1966), and used in e.g. produc-
tion of viscose rayon. However, we have found no previous publi-
cation on alkaline pretreatment of waste textiles and enzymatic
hydrolysis.

0960-8524/$ - see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2008.07.020
1008 A. Jeihanipour, M.J. Taherzadeh / Bioresource Technology 100 (2009) 1007–1010
The aim of this work was to investigate the possibility of com-
plete conversion of waste cotton-based textiles to ethanol via
enzymatic hydrolysis and fermentation. However, since the yield
in enzymatic hydrolysis of untreated cotton was low in the preli-
minary experiments, the main effort was to develop a suitable pre-
treatment process for the cotton textiles.
2. Methods
2.1. Substrates, enzymes and yeast strain
The cotton-based materials used in this work were cotton linter
(purchased from a local shop in Sweden) and used blue jeans tex-
tiles. Pure cellulose (Avicel, Fluka 11365) was used as a reference
material for comparison. The cellulose content of samples was
determined according to Ruiz and Ehrman (1996). Two enzymes,
cellulase (SIGMA, C2730) and b-glucosidase (SIGMA, G0395), were
used for enzymatic hydrolyses. The activity of cellulase was mea-
sured as 60 FPU/mL of enzyme solution according to Adney and Ba-
ker (1996), while the activity of b-glucosidase was reported as
5.2 IU/mg solid by the supplier. Flocculating yeast strain of Saccha-
romyces cerevisiae CCUG 53310 (Culture Collection, University of
Göteborg, Sweden) was used in all the cultivation experiments.
The strain was maintained on agar plates made of yeast extract
10 g/L, soy peptone 20 g/L, and agar 20 g/L with D-glucose 20 g/L
as an additional carbon source.
2.2. Pretreatment procedures
In this investigation, cotton linter was soaked within 3 h in
various concentrations of sodium hydroxide solution (0%, 2%,
4%, 6%, 8%, 10%, 12%, 14%, 16%, 18% and 20%) at three different
temperatures of 0 °C, 23 °C and 100 °C. The ratio of liquid/dry so-
lid was 20 mL/g. The chemically treated fibers were then washed
with distilled water until pH 7 was obtained. The washed sam-
ples were squeezed to remove the water and kept in a refriger-
ator for further use. Jeans textiles were pretreated at the best
conditions with the same procedure as for the cotton fibers.
But before pretreatment of the jeans material, it was shredded
and then ground to small size. Pretreatment of the textiles with
concentrated phosphoric acid was performed according to Zhang
et al. (2007) with some modifications. The dry weight of the
treated and untreated materials was measured by drying at
110 °C overnight.
2.3. Digestibility, saccharification and fermentation
Digestibility or saccharification of cotton, jeans or Avicel as a
reference were investigate by enzymatic hydrolyses at 45 °C and
pH 4.8 (with 50 mM sodium citrate buffer) with 3% w/v solid sub-
strates, using 20 FPU cellulase and 30 IU b-glucosidase per gram
cellulose within 24–96 h. The hydrolyzate or glucose solution were
then supplemented with other nutrients (Karimi et al., 2006), inoc-
ulated with 10 ± 3 g/L S. cerevisiae, and cultivated at 37 °C for 4
days. The yields for pretreated materials were calculated based
on the amount of retrieved cellulose after pretreatment. All exper-
iments were performed at least in duplicate and results are pre-
sented as average values. The average standard deviation of the
results was 4.0%.
2.4. Analytical methods
The samples from various steps of experiments were analyzed
by HPLC (Waters, Milford, USA), which was equipped with RI
detector (Waters 2414). Glucose and ethanol were analyzed on
an Aminex HPX-87H column (Bio-Rad, USA) at 60 °C with 0.6 mL/
min eluent of 5 mM sulfuric acid.
3. Results
3.1. Alkali pretreatment at different temperatures and concentrations
Conversion of cotton-based waste textiles to ethanol by enzy-
matic degradation and fermentation was investigated. The materi-
als used were cotton linter and a waste material from blue jeans
textiles. The cellulose contents of these materials were 98% ± 1%
and 93% ± 1%, respectively. The cotton and jeans with no pretreat-
ment were enzymatically hydrolyzed to glucose by cellulase and b-
glucosidase for 2 days, and resulted into 24.1% and 23.6% digestion,
respectively. Hydrolysis of pure cellulose (Avicel) at identical con-
ditions was more successful, where 38.7% of the cellulose was con-
verted to glucose.
Pretreatment of the textile materials by concentrated phospho-
ric acid improved their digestibility. The cellulose parts of the
materials were completely dissolved in the acid, and then precipi-
tated using acetone, where 70–80% of the initial materials were
recovered after the precipitation. It resulted in improved glucose
yield in the enzymatic hydrolyses, where 63.4% and 60.7% of the
cotton linter and the jeans were digested, respectively. However,
better results obtained by the alkali pretreatment of the cotton
and jeans (data not shown).
Cotton linter was then chosen as a sample of cotton textiles to
study the effect of temperature and NaOH concentration on the
enzymatic digestibility of the material. The material was treated
at 0 °C, 23 °C and 100 °C with 0–20% NaOH for 3 h. The treated
materials remained solid in the solution, but swelled, where more
than 95% of the initial materials were recovered after the washing.
The most important results of digestion of pretreated cottons are
summarized in Table 1. The results show great effects of treatment
temperature and NaOH concentration on the rate of enzymatic
hydrolysis. The best results were obtained at cold temperature
(0 °C), where more than 85% of the theoretical glucose yield could
be obtained within 24 h enzymatic hydrolysis of treated cotton by
12% NaOH solution (Table 1). Furthermore, increasing the concen-
tration of NaOH enhanced the digestibility. However, for pretreat-
ment at 0 °C, the concentration of 12% was the critical dosage and
no significant improvement in digestibility was obtained when
using higher concentration of NaOH. Temperature had a negative
impact on the rate of enzymatic hydrolysis. The initial enzymatic
hydrolysis rate during the first day was practically the same for
all three temperatures until 8% NaOH concentration used (Table
1). However, at higher concentration of sodium hydroxide used,
lower temperature was more effective for increasing the hydrolysis
rate. The best digestion results were obtained at 12% NaOH and
0 °C, where 99.1% of the cotton was digested to glucose within 4
days.
3.2. Ethanol production from waste textiles
Production of ethanol from cotton linter and waste blue jeans
textiles was investigated and the results were compared with Avi-
cel and glucose as reference materials. The textiles were untreated
or treated either with 12% NaOH at 0 °C for 3 h or with concen-
trated phosphoric acid. The materials were first hydrolyzed at
45 °C by the enzymes for 24 h followed by simultaneous sacchari-
fication and fermentation (SSF) by S. cerevisiae for 4 days. The most
important results are summarized in Fig. 1. As the reference mate-
rial, the yields of ethanol from glucose after 24 to 96 h cultivations
were 0.42 and 0.44 g/g, which was 83% and 86% of the theoretical
yield, respectively. The SSF of untreated Avicel resulted in 35% and
 A. Jeihanipour, M.J. Taherzadeh / Bioresource Technology 100 (2009) 1007–1010 1009

Table 1
Digestion (percentage of conversion) of pretreated cotton linter to glucose by enzymatic hydrolysis in 24, 48 and 96 h

NaOH concentration (%w/v) Pretreatment at 0 °C Pretreatment at 23 °C Pretreatment at 100 °C

Hydrolysis time Hydrolysis time Hydrolysis time
24 h 48 h 96 h 24 h 48 h 96 h 24 h 48 h 96 h
0 23.2 42.7 44.7 17.6 24.1 26.2 17.6 24.1 26.2
2 25.6 35.8 42.2 18.8 28.1 30.1 18.8 28.1 30.1
4 29.4 40.6 52.0 20.3 30.3 36.0 20.3 30.3 36.0
6 40.4 43.0 57.8 32.3 42.3 49.1 32.3 42.3 49.1
8 42.6 44.1 58.9 43.7 44.0 52.1 43.7 44.0 52.1
10 52.3 70.5 81.6 47.2 55.3 59.0 47.2 55.3 59.0
12 85.1 93.3 99.1 50.4 75.5 93.1 50.4 75.5 93.1
14 86.4 94.1 98.9 50.0 76.5 98.8 50.0 76.5 98.7
16 88.0 94.7 97.9 69.4 80.2 98.8 69.4 80.2 98.8
18 86.7 93.1 98.7 79.9 88.0 99.1 79.9 88.0 99.1
20 78.5 83.6 97.3 77.4 89.7 98.6 77.4 89.7 98.6

The pretreatments were carried out in 3 h at 0 °C, 23 °C and 100 °C with 0–20% NaOH. The enzymatic hydrolyses were performed at 45 °C using 30 g/L cotton, 20 FPU/g
cellulase and 30 IU/g b-glucosidase.

100 0.48 g ethanol from each gram of the pretreated waste textiles.
90 Furthermore, the hydrolysis of the material was quite successful,
Ethanol yield (% theoretical)

so that with just 24 h SSF the yield of ethanol from the treated tex-
80 tiles was 92% of the corresponding yield from glucose.
70 The pretreatment of the textiles by phosphoric acid was not as
effective as the alkali pretreatment. The acid-pretreated cotton and
60
jeans resulted in 44–46% of the theoretical ethanol yield after one
50 day and 65–66% after 4 days of SSF (Fig. 1b). However, this is still
40
130–160% improvement compared to the untreated textiles.

30
4. Discussion
20
Almost one third of the waste textiles produced from manufac-
10
turing clothing and other textile products are made of cotton. The
0 overall objective of this research was to develop an efficient meth-
od for bioconversion of this type of waste in an environmentally
100 safe manner. The current work shows the possibility of converting
90 the cotton-based waste textiles to glucose with more than 99%
Ethanol yield (% theoretical)

yield and a total ethanol production of 0.48 g/g of the pretreated

80 cotton.
70 An advantage of cottons compared to the other lignocelluloses
is not covering the glucan polymers with lignin and hemicellu-
60
loses. Therefore, one might consider easy digestion of cottons with
50 cellulases. However, the highly crystalline structure of cotton
40 makes it resistant to the enzymes. NaOH pretreatment influences
the structure and morphology of cellulose fibers. The most impor-
30 tant effect of the base is the crystallographical modification of the
20 fibers from cellulose I to cellulose II (Kleinschek et al., 1999), which
has a major impact on the digestibility of the cottons. Phosphoric
10
acid was also able to improve this characteristic, but not as much
0 as NaOH did.
0 24 48 72 96 120
Treatment with sodium hydroxide at cold temperature has been
Time (h) studied for dissolution of cellulose, where the dissolution can take
Fig. 1. The trend of ethanol production in 24 h enzymatic hydrolysis at 45 °C
place in a narrow range of NaOH concentration, from 7% to 9% w/v,
followed by a 4 days SSF at 37 °C. The symbols represent (a) glucose (x) and Avicel and temperature, 5 °C to 6 °C (Cai and Zhang, 2006; Isogai and
(d) as reference materials, untreated cotton (N) and waste jeans textiles (j) and Atalla, 1998; Roy et al., 2001). The conditions that we examined in
(b) pretreated cotton (N) and waste jeans textiles (j) with NaOH, pretreated cotton this work were out of this range. However, the pretreatment at
(4) and waste jeans textiles (h) with concentrated phosphoric acid.
cold conditions was more successful. According to Mansour et al.
(1972), during the alkaline treatments, crystallinity is reduced
55% of the theoretical yield of ethanol in one and four days, respec- most in presence of 9–12% NaOH, which is in agreement with
tively (Fig. 1a). our results. This pretreatment of cotton stretches some of the
The results show significant improvement in ethanol produc- hydrogen bonds beyond the breaking limit. But on washing with
tion from the cotton and jeans by the alkali pretreatment. While water, reformation of bonds occurs with various strengths
the SSF of untreated jeans and cotton resulted in 25–26% of the (Mansour et al., 1972). Therefore, some of the hydroxyl groups
theoretical ethanol yield within 4 days (Fig. 1a), the alkali treat- may enter into stronger hydrogen bonds, while others enter into
ment resulted in 85–86% of the theoretical yield which is the same weaker ones, with the overall number of the hydroxyl groups
as the yield from glucose (Fig. 1b). This means that we produced entering the strong bonds being lower than in the original
1010 A. Jeihanipour, M.J. Taherzadeh / Bioresource Technology 100 (2009) 1007–1010
untreated samples. The effect of temperature in this process might
be connected with the stronger hydrogen bonding and network of
NaOH hydrates at lower temperature, which facilitates penetration
of NaOH hydrates into the cellulose fibers (Egal et al., 2007). This
fact was investigated and confirmed by comparing the spectra re-
corded at 5 °C and room temperature (Kleinschek et al., 1999).
These phenomena might explain why lower temperature has a
higher impact on the enzymatic hydrolysis of cotton.
5. Conclusion
We may infer the possibility of converting cotton-based waste
materials to ethanol via enzymatic hydrolysis and fermentation.
However, they need a pretreatment; otherwise the ethanol yield
cannot be more than 26% of the theoretical yield. Pretreatment
with phosphoric acid results in a great improvement in the hydro-
lysis of the materials, but it is far from the results of alkali pretreat-
ment by NaOH, which results in more than 99% yield in the
subsequent enzymatic hydrolysis.
Acknowledgements
This work was financed by Swedish Excellent Center ‘‘Waste
Refinery”. The authors also appreciate collaboration and discus-
sions with SP Technical Research Institute of Sweden, Borås Energi
& Miljö AB and Dalkia FM AB in Sweden.
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