Bioprocess Biosyst Eng (2007) 30:189–196
DOI 10.1007/s00449-007-0114-3
ORIGINAL PAPER
Bioconversion of municipal solid waste to glucose for bio-ethanol
production
Aiduan Li Æ Blanca Antizar-Ladislao Æ
Majeda Khraisheh
Received: 11 December 2006 / Accepted: 10 January 2007 / Published online: 15 February 2007

Springer-Verlag 2007
Abstract Selected biodegradable municipal solid
waste fractions were subjected to fifteen different pre-
hydrolysis treatments to obtain the highest glucose
yield for bio-ethanol production. Pre-hydrolysis treat-
ments consisted of dilute acid (H2SO4, HNO3 or HCl, 1
and 4%, 180 min, 60C), steam treatment (121 and
134C, 15 min), microwave treatment (700 W, 2 min)
or a combination of two of them. Enzymatic hydrolysis
was carried out with Trichoderma reesei and Tricho-
derma viride (10 and 60 FPU g–1 substrate). Glucose
yields were compared using a factorial experimental
design. The highest glucose yield (72.80%) was ob-
tained with a pre-hydrolysis treatment consisting of
H2SO4 at 1% concentration, followed by steam treat-
ment at 121C, and enzymatic hydrolysis with Tricho-
derma viride at 60 FPU g–1 substrate. The contribution
of enzyme loading and acid concentration was signifi-
A. Li
B. Antizar-Ladislao
M. Khraisheh
Department of Civil and Environmental Engineering,
University College London, Gower Street,
London WC1E 6BT, UK
A. Li
e-mail: aiduan.li@ucl.ac.uk
M. Khraisheh
e-mail: m.khraisheh@ucl.ac.uk
Present Address:
B. Antizar-Ladislao (&)
Department of Water and Environmental Science
and Technology, Universidad de Cantabria,
Bulevard Ronda Rufino Peón, 254,
39316 Torrelavega, Cantabria, Spain
e-mail: antizarb@unican.es
cantly higher (49.39 and 47.70%, respectively), than
the contribution of temperature during steam treat-
ment (0.13%) to the glucose yield.
Keywords Bioethanol
Glucose
Lignocellulose

Municipal solid waste
Introduction
The use of biofuels for transport is becoming of
increasing importance for a number of reasons, such
as environmental concerns relating to climate change,
depletion of fossil fuel reserves, and reduction of
reliance on imports [1, 2]. This is leading to inter-
national, national and regional focus upon alternative
energy sources. In Europe, the European Commis-
sion has proposed indicative targets for biofuel sub-
stitution of 5.75% by 2010 [1]. A potential source for
low-cost biofuel (i.e., bio-ethanol) production is to
utilize lignocellulosic materials such as crop residues,
grasses, sawdust, wood chips, and solid waste. Addi-
tionally, European legislative pressures target for
minimising landfill use in European countries, and
the amount of biodegradable municipal solid waste
(BMSW) going to landfill must be reduced by 25%
by 2010, 50% by 2013 and 65% by 2020 [1]. Thus, the
BMSW fraction may be considered an alternative
sustainable source of bio-ethanol [3, 4]. In fact,
London Boroughs (UK) collected 4.4 thousand ton-
nes of total municipal solid waste (MSW) in 2004–
2005 [5]. Although the recycling/composting rate in
London has increased over the past 5 years from 8%
in 2000–2001 to 15% in 2004–2005, still 65% MSW is
disposed by landfill, of which 81.4% may be biode-
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gradable to a given extent, averaging 65% biode-
gradability [5, 6]. Typical BMSW fractions are paper/
card, kitchen waste, garden waste, textiles, fines and
miscellaneous (combustibles and others).
In order to satisfy European legislative pressures
and energy demand, an environmentally sustainable
approach required to change the current waste man-
agement strategies will be the bioconversion of ligno-
cellulosic BMSW fraction-to-bio-ethanol. A limited
number of studies on the application of this emerging
sustainable approach have been reported [4]. Previous
research in bioconversion of lignocellulosic materials
(i.e., agricultural residues, woods, residues from pulp
and paper industry, urban lignocellulosic wastes) to
bio-ethanol in the last two decades due to its large
availability and immense potential clearly indicate that
in order to obtain high glucose yields, removal of lignin
and hemicellulose (hinders the access of enzymes to
cellulose), reduction of cellulose crystallinity, and in-
crease of porosity in pre-treatment processes can sig-
nificantly improve the hydrolysis of lignocellulosic
materials [7].
For that purpose, physical, physico-chemical, chem-
ical and biological pre-treatment processes have been
applied to several cellulosic materials to enhance their
enzymatic digestibility [8]. Mtui and Nakamura [4] ap-
plied a pre-hydrolysis stage (carried out with dilute
strong acid followed by steam treatment at 120C for
15 min) and enzymatic hydrolysis (carried out with
cellulase enzyme extracted from Trichoderma reesei,
incubated at 55C for 6 h) to lignocellulosic solid wastes
from selected sites in Tanzania. They achieved a glucose
concentration of 0.13 and 0.05 g l–1 (corresponding to
1 g of pre-treated lignocellulosic material) from solid
waste samples with a high lignocellulose content (93%)
and a low lignocellulose content (14%), respectively.
Higher glucose concentration (and yields) needs to be
obtained in order to implement BMSW-to-bio-ethanol
as a feasible environmental sustainable waste manage-
ment strategy.
Pre-hydrolysis treatment and enzymatic hydrolysis
of BMSW are of crucial importance during the bio-
conversion of BMSW-to-bio-ethanol, and thus their
optimization will result in beneficial environmental and
economic practices [9]. Hence, this research sought to
investigate the impact of pre-hydrolysis treatments
(i.e., dilute acid type and concentration, steam treat-
ment, microwave) and enzymatic hydrolysis (i.e., en-
zyme type and concentration) on the bioconversion of
BMSW-to-glucose at laboratory-scale to optimize the
processes for bio-ethanol production and bio-ethanol
technology.
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Bioprocess Biosyst Eng (2007) 30:189–196
Materials and methods
Waste selection and physical pre-treatment
Waste feedstock was selected from typical biodegrad-
able MSW fractions, which are carrot peelings (CP)
and potato peelings (PP) typical in kitchen waste, grass
(G) typical in garden waste and newspaper (NP) and
scrap paper (SP) typical of paper/card fractions. Potato
and carrot were peeled separately. The peelings, grass,
newspaper and scrap paper were separately shredded
(2–20 mm) using a food processor (Magimix UK Ltd)
and dried at 105C overnight. Waste fractions were
then separately grinded, screened (0.2 mm) and stored
in a dark and dry environment at room temperature
previous to treatment.
Pre-hydrolysis treatment
Various pre-hydrolysis treatments were investigated,
including dilute-acid pre-hydrolysis, steam treatment
and microwave treatment or combinations of two of
them applied consecutively (Table 1). The effect of
sulphuric acid (H2SO4), nitric acid (HNO3), and
hydrochloric acid (HCl) was investigated in this study.
Biomass and dilute acid (4%) were mixed at a solid/
liquid ratio of 0.5 g in 50 ml during 180 min at 60C.
The acid-soaked samples were drained and air-dried
overnight to a final 45% moisture content.
Additionally, two different pre-treatment methods
were combined by treating the BMSW fraction with
4% H2SO4, HNO3, HCl, for 180 min at 60C first fol-
lowed by steam treatment. Steam treatment was
undertaken in an autoclave at a constant temperature
(121 or 134C) for 15 min. Following steam treatment,
the samples were washed with deionised water until
the pH of the filtrate remained constant at an
approximate value of 4.5. The pre-treated samples
were drained and air-dried overnight to a final 45%
moisture content. Then, the dilute acid treatment fol-
lowed by steam treatment that produced the highest
glucose yield was compared with acid treatment fol-
lowed by microwave treatment. Microwave treatment
was carried out in a domestic microwave oven (Matsui
TS106WH) at full power output of 700 W for 2 min
[10]. Biomass and dilute acid (4%) were mixed at a
solid/liquid ratio of 0.5 g with 4% acid in a 100 ml open
beaker and placed in the centre of a rotating circular
glass plate in the microwave. The idea was to speed the
reaction at high temperature inside the microwave
oven and to save time in comparison with the steam
Bioprocess Biosyst Eng (2007) 30:189–196 191

Table 1 Experimental conditions in pre-hydrolysis treatment of BMSW fractions selected in this study
Experiment Dilute acid Steam treatment Microwave treatment Enzymatic
hydrolysis
HNO3 Time, HCl Time H2SO4 Time T Time Power Time Cellulase
(%) min (%) (min) (%) (min) (C) (min) (W) (min) (FPU/g)

1 4 180 – – – – – – – – 60
2 – – 4 180 – – – – – – 60
3 – – – – 4 180 – – – – 60
4 4 180 – – – 121 15 – – 60
5 – – 4 180 – – 121 15 – – 60
6 – – – – 4 180 121 15 – – 60
7 – – – – 4 180 – – 700 2 60
8 – – – – 4 180 134 15 – – 60
9 – – – – 1 180 121 15 – – 60
10 – – – – 1 180 134 15 – – 60
11 – – – – 4 180 121 15 – – 10
12 – – – – 4 180 134 15 – – 10
13 – – – – 1 180 121 15 – – 10
14 – – – – 1 180 134 15 – – 10
15a 4 180 121 15 – – 60
a
Enzyme used is T. reesei

treatment, which lasted for 50 min. A few bubbles
started to appear on the surface of the material indi-
cating the onset of boiling. Following microwave
treatment, the samples were washed with deionised
water until the pH of the filtrate remained constant
pH = 4.5. The pre-hydrolysis treated samples were
drained and air-dried overnight to a final 45% moisture
content.
Enzymatic hydrolysis
Enzymatic hydrolysis of 0.5 g of pre-hydrolysed sub-
strate was carried out at 50C in vials (50 ml) placed in
an orbital agitator at 68 rpm for 96 h. The liquor pH
was kept at pH = 4.8 using 0.1 M citrate acid-sodium
citrate buffer. Two enzymes, T. viride and T. reesei
(Cellulase from T. viride and cellulase from T. reesei
ATCC26921, respectively; Sigma, UK) were separately
used as received in combination with the enzyme b-
glucosidase (Sigma, UK). Possible differences in the
cellulase activity of the enzymes [11] was measured by
the Filter Paper assay developed by Mandels et al. [12],
and the activity was expressed in terms of Filter Paper
Units (FPU). The b-glucosidase activity was measured
by the PNPG assay [13], and reported as PNPG units
(PNPGU). All the experiments were performed at
enzyme to substrate ratios of 60 FPU g–1 substrate [14]
and 64 PNPGU g–1 substrate, and 10 mg ml–1 puro-
mycin for 96 h. Samples were withdrawn from the
reaction media after 24 h for pre-hydrolysis treatment
comparison and at 2, 5, 9, 24, 48, 72, and 96 h for ki-
netic studies.
Analytical methods
The analytical methods were performed according to
the following procedures. Ash content was determined
using the Standard Biomass Analytical Methods [15].
Briefly, triplicate 10 g samples were dried at 105C for
24 h (moisture content) and then transferred to a
muffle furnace at 575C for 24 h. The cellulose content
of the selected biodegradable MSW fractions was
analyzed by the method of Crampton and Maynard
[16]. Sugar concentrations were determined routinely
from centrifuged samples (13,000 rpm, 5 min) using a
glucose meter (MediSense Ltd, UK) after the method
was validated in the laboratory. Calibration graphs
were linear for the concentration range 0.5–5 g l–1
(R2 = 0.98).
Data analysis
All compositional analyses were done in triplicate, and
data were corrected to a 100% dry matter (DM) basis.
Glucose yield as percentage of the theoretical yield
[percentage digestibility, obtained from the equation
which involves the transfer of cellulose to sugar
(C6H10O5)n + nH2O fi (C6H12O6)n] was computed
by using the formula given by the National Renewable
Energy Laboratory (Standard Biomass Analytical
Procedures) [17]:

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Glucose yield ð%Þ ¼ ½Glucose=ð1:11  f
 ½BiomassÞ  100
where [Glucose] is the glucose concentration (g l ), –1
[Biomass] is dry biomass concentration following
enzymatic hydrolysis (g l–1) = 10 g l–1; f is the cellu-
lose fraction in dry biomass (g g–1) (Table 2); and
1.11 is the factor that corresponds to the mass bal-
ance of the transfer of cellulose to sugar. Glucose
yields following different treatments in this study
were compared using a factorial experimental design
with the software package Design Ease 6.0.6 (State-
ease, Inc.). Selected factors identified as more con-
venient to control the hydrolysis process were se-
lected from both, pre-hydrolysis treatment and
enzymatic hydrolysis. Thus, factors included pre-
hydrolysis temperature, pre-hydrolysis acid concen-
tration and cellulase concentration during enzymatic
hydrolysis (Table 1).
Results and discussion
Composition of the MSW fractions
Composition of the waste fractions is summarised in
Table 2. Experimental results show that about 40, 19,
22, 46 and 67% of the oven-dry CP, PP, G, NP and SP
weight, respectively, corresponds to cellulose. Ligno-
cellulosic biomass typically contains 55–75% carbohy-
drates (including cellulose) by dry weight, for example
cardboard, Eucalyptus globules and Pinus pinaster
contain 75, 62.3 and 59.6% carbohydrates by dry weight,
respectively [18, 19]. Although the cellulose content of
some BMSW fractions may be low as compared to other
lignocellulosic biomass, the optimization of pre-hydro-
lysis treatments of BMSW fractions and mixtures may
result in glucose yields similar to those currently re-
ported for agricultural waste, rich in lignocellulosic
biomass. Thus, pre-hydrolysis treatments need to be
optimised.
Table 2 Composition of BMSW fractions selected in this study
Fraction of MSW Oven-dry basis (wt %)
Cellulose [27] (f) Ash content
Carrot peelings 39.49 9.71
Potato peelings 18.76 9.04
Grass 21.60 16.57
News paper 46.12 6.78
Scrap paper 67.07 16.32
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Bioprocess Biosyst Eng (2007) 30:189–196
Pre-hydrolysis treatment of the MSW fractions
Dilute acid hydrolysis has been successfully developed
at low temperatures (T less than 160C) and high solids
loading (10–40%) favourable for cellulose hydrolysis
and high reaction rates [20]. In order to asses the
suitability of each pre-hydrolysis treatment for maxi-
mum solubilization of hemicelluloses and subsequent
enzymatic hydrolysis, the effect of pre-hydrolysis
treatment of BMSW with the acid was investigated
using H2SO4, HNO3 and HCl (4%). Grass resulted in
the highest glucose yield (42–49%), while CP, PP, SP
resulted in lower, although similar, glucose yields (15–
29%) for all dilute acid pre-hydrolysis treatments. NP
resulted in the lowest glucose yields (5–10%). In gen-
eral, the pre-hydrolysis treatment for the selected
BMSW fractions with H2SO4 resulted in similar or
higher glucose yields than with HNO3 or HCl (Fig. 1).
Although dilute acid pre-hydrolysis treatment can
significantly improve the cellulose hydrolysis, its cost is
usually higher than some physico-chemical pre-hydro-
lysis treatments such as steam treatment or microwave
treatment. Steam treatment, most commonly used
method for pre-treatment of lignocellulosic materials,
causes hemicellulose degradation and lignin transfor-
mation due to high temperature, thus increasing the
potential of cellulose hydrolysis. Low temperature and
longer residence time are favourable for optimal
hemicellulose solubilisation and hydrolysis [8]. Addi-
tion of dilute acid in steam treatment can effectively
improve enzymatic hydrolysis, decrease the production
of inhibitory compounds, and lead to more complete
removal of hemicellulose [21]. Microwave method can
also be applied to improve acid hydrolysis [22]. Thus in
this study the pre-hydrolysis treatment of dilute acid
(H2SO4, HNO3 and HCl) followed by steam treatment
PP+CP+G+SP+NP
CP+PP H2SO4
SP+NP HNO3
HCl
SP
NP
G
PP
CP
0 20 40 60 80 100
Glucose yield, %
Fig. 1 Effects of pre-hydrolysis treatments of selected BMSW
fractions in glucose yield after 24 h hydrolysis (dilute acid
concentration 4%, T.viride 60 FPU g–1). CP carrot peelings, PP
potato peelings, G grass, NP newspaper, SP scrap paper
Bioprocess Biosyst Eng (2007) 30:189–196
to effectively maximize enzymatic hydrolysis was
investigated. Subsequently, the best experimental
conditions were compared with dilute acid treatment
followed by microwave treatment.
The effective pre-hydrolysis treatment for the se-
lected BMSW fractions in this study occurred com-
bining dilute H2SO4 (4%) followed by steam treatment
(Fig. 2). Highest glucose yield after 24 h pre-hydrolysis
treatment was obtained from grass and carrot peelings
(62 and 60%, respectively), followed by potato peel-
ings (32%). The glucose yield from waste paper was
relatively lower, where the scrap paper and newspaper
resulted in 25 and 6% glucose yield, respectively.
Previous studies have reported that softwood newspa-
per is the most difficult component to digest in a lig-
nocellulosic MSW mixture, due to its high lignin
content [23]. In this study, no pre-treatment or com-
bining dilute HCl with steam treatment resulted in the
highest glucose yields from scrap paper and newspaper
(32–46% scrap paper and 18% newspaper) as com-
pared to other pre-hydrolysis treatments. Comparing
the combined treatments H2SO4 with steam treatment
and H2SO4 with microwave, generally higher glucose
yields were obtained using the former treatment, but
the glucose yield from newspaper was doubled using
H2SO4 followed by microwave pre-hydrolysis treat-
ment.
Glucose yields following the combined treatment of
dilute acid with microwave and the glucose yield fol-
lowing the same dilute acid treatment alone were
compared (Figs. 1, 2). Thus the effect of microwave
treatment following dilute acid treatment could be
revealed. It was observed that the glucose yield was
generally lower after microwave treatment. It was
SP
NP H2SO4+MW
H2SO4+SE
G HNO3+SE
HCl+SE
no pretreatment
PP
CP
0 20 40 60 80 100
Glucose yield, %
Fig. 2 Effects of combined pre-hydrolysis treatments of selected
BMSW fractions in glucose yield after 24 h hydrolysis (dilute
acid concentration 4%, T.viride 60 FPU g–1). CP carrot peelings,
PP potato peelings, G grass, NP newspaper, SP scrap paper
193
suspected that although microwave treatment may be
more efficient in removing hemicellulose and lignin
present in the selected BMSW fractions and therefore
enabling the accessibility of enzymes to the cellulosic
chains, it also may have facilitated glucose production
at concentrations, which may have been inhibitory
during enzymatic hydrolysis. However, more research
is currently undertaken in order to provide with a
rigorous explanation to this observation.
Pre-hydrolysis treatment methods investigated in
this study indicated that in general lower glucose yield
resulted from newspaper and scrap paper BMSW
fractions than from kitchen waste BMSW fractions,
i.e., carrot peelings and potato peelings. Nguyen and
Saddler [14] reported a maximum glucose yield
approximately 60% after two-stage dilute-acid pre-
treatment of softwood, which is the source for news-
paper and scrap paper. A lower glucose yield obtained
from newspaper and scrap paper in this study than by
Nguyen and Saddler [14] may have occurred due to the
dense structure and presence of chemicals, such as
fillers, ink and other additives in the newspaper and
scrap paper (biosurfactants and antimicrobials to block
pulp degradation), which may have inhibited the
enzymatic hydrolysis [24].
Enzymatic hydrolysis of MSW fractions
Efficient hydrolysis of cellulose requires the synergistic
activities of three types of enzymes. Endo-b–1,4-glu-
canases hydrolyze accessible regions on cellulose
chains to provide new sites for attack by exo-acting
cellobiohydrolases, which removes successive cellobi-
ose units from newly created chains ends. Finally, b-
glucosidase hydrolyzes cellobiose, and smaller amounts
of higher cellooligomers, to glucose. Two cellulose-
digesting enzymes used in this study T. reesei and T.
viride contain endogenous b-glucosidase activity,
however, the activities of this enzyme are generally
insufficient to prevent the accumulation of cellobiose,
resulting in product inhibition of endoglucanases and
cellobiohydrolases [25]. Consequently, b-glucosidase
was supplemented during the enzymatic hydrolysis of
MSW fractions.
The substrates included single substrate from the
selected MSW fractions (carrot peeling, potato peel-
ings, grass, newspaper and scrap paper) and multi-
substrates by combining different single substrates. All
substrates were pre-hydrolysis treated combining di-
lute acid (H2SO4, 4%) followed by steam treatment, as
this was shown the most efficient pre-hydrolysis treat-
ment for MSW fractions (Fig. 3). This study indicated
that using T. viride resulted in higher glucose yields
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194 Bioprocess Biosyst Eng (2007) 30:189–196

PP+CP+G+SP+NP for the reduction of the glucose yield in the selected

CP+PP waste mixture. Thus, future efforts need to be oriented
SP+NP to increase glucose yields from newspaper waste and
SP T. reesei mixtures of BMSW containing newspaper.
T. viride
NP

G Process optimization
PP

CP
Avicel
0 20 40 60 80
Glucose yield, %
Fig. 3 Effects of cellulase (T. viride, T. reesei, 60 FPU g–1)
during enzymatic hydrolysis of selected BMSW fractions in
glucose yield (55C, 96 h). Pre-hydrolysis treatment: H2SO4 (4%,
180 min) and steam treatment (121C, 15 min). CP carrot
peelings, PP potato peelings, G grass, NP newspaper, SP scrap
paper
than using T. reesei, which corroborates previous
studies indicating an activity (units glucose produced
per mg enzyme) of T. viride and T. reesei of 3–10 and
1.0, respectively [26].
Once pre-hydrolysis treatment (H2SO4 4% and
steam treatment) and cellulose-digesting enzymes (T.
viride) were optimized for maximum glucose yield
from selected BMSW fractions, a set of experiments of
enzymatic hydrolysis was carried out during 96 h, and
samples were withdrawn at intervals (Fig. 4). This ki-
netic study indicated that the highest glucose yield was
obtained using potato peeling (89%), followed by
carrot peelings (73%) or combination of carrot peel-
ings and potato peelings (70%). Newspaper produced
the lowest glucose yield (14%) following 96 h enzy-
matic hydrolysis. The glucose yield produced by a
combination of five selected BMSW fractions (20%
each) increased from 41 to 52% after 24 and 96 h of
enzymatic hydrolysis, respectively. The presence of
newspaper was suspected to be the main responsible
Mathematical modelling is a valuable tool for the
process optimization, and thus various studies have
developed mathematical models considering several
100
factors. For example, Nguyen and Saddler [14] used
and exponential function, R0 ¼ t  eðTr Tb Þ =14:75, to
optimize the pre-treatment process including two fac-
tors (temperature and residence time) and the process
was optimized with different severities by changing the
temperature. Where, R0 is the severity factor combin-
ing the effects of time and temperature, t is the resi-
dence time (min), Tr is the reaction temperature (C),
and Tb is a reference temperature (100C). Esteghla-
lian et al. [20] modelled the hydrolysis of hemicellulose
to its monomeric constituents and possible degradation
of these monomers by a series of first-order kinetic
functions. The kinetic parameters for predicting the
xylose yield were determined using the actual acid
concentration in the reactor after accounting for the
neutralization effect of the substrates. Yañez et al. [18]
used a pseudo-homogeneous first order kinetic func-
tion to calculate the hemicellulose (H) and non-poly-
saccharide (NPF) fractions and an overall material
balance to estimate the cellulose content (Cl),
Cl = 100 – H – NPF. These studies have facilitated
process optimization, however, current technology
development for the production of bio-ethanol focus
towards the utilisation of residual lignocellulosic
materials at lower production costs. Thus, important
factors that need to be optimized in order to reduce the
cost of the production of bio-ethanol include acid
concentration, temperature and cellulase concentra-
tion.

Fig. 4 Kinetics of glucose 100

Avicel
yield following pre-hydrolysis
treatment with H2SO4 (4%) 80
PP
and steam treatment (121C,
Gluc os e y iedl, %

CP
15 min) and enzymatic
60 G
hydrolysis with T. viride
(60 FPU g–1). CP carrot SP
peelings, PP potato peelings, 40
G grass, NP newspaper, SP NP
scrap paper 20 CP+PP
NP+SP
0
0 20 40 60 80 100 C+P+G+NP+SP
Time, h

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Bioprocess Biosyst Eng (2007) 30:189–196 195

Thus, once the most effective pre-hydrolysis and
enzymatic hydrolysis for the selected BMSW fractions
were identified in this study, the hydrolysis process was
optimized by using a factorial experimental design
considering the aforementioned factors. Glucose yields
using pre-hydrolysis treatment of carrot peelings and
potato peelings (50% each), adding dilute H2SO4 acid
(1 and 4%; 180 min), steam treatment (T, 121 and
134C; 15 min), and T. viride (10 and 60 FPU g–1
substrate) following hydrolysis at 50C for 72 h were
computed using the software package Design Ease
6.0.6 to find the optimal treatment conditions (Fig. 5).
The highest glucose yield (72.80%) was obtained at
dilute acid concentration (1%), low temperature
(121C) and high enzyme loading (60 FPU g–1 sub-
strate). The contribution of enzyme loading and acid
concentration to the glucose yield was significantly
higher (49.39 and 47.70%, respectively), than the con-
tribution of temperature during steam treatment
(0.13%). Additionally, the results obtained by the
model presented a satisfactory correlation to the actual
values (R2 = 0.90).
The use of dilute H2SO4 acid concentration is an
advantage, in particular since its cost is usually higher
than the cost of using other physico-chemical pre-
hydrolysis treatments [8, 9]. Additionally, this study
indicated that lower temperature (121C) and long
residence time (96 h) during steam treatment produced
the highest glucose yields, which additionally has been
recognized as one of the most cost-effective pre-
hydrolysis treatment process for agricultural residues [9,
27]. Neutralization of pH is necessary for the down-
stream enzymatic hydrolysis or fermentation process,
which may be avoided using low H2SO4 acid concen-
tration (1%). An optimal ratio of cellulase to cellulose
0.8 FPU g–1 substrate has been previously reported
using Avicel pH 105 and T. reesei [28], while a ratio
0.2 FPU g–1 substrate was reported to be inhibitory [29].
This study indicated that BMSW fractions required
higher cellulase to cellulose ratios to obtain high glucose
yields, which will increase the cost of the process.
Conclusions
Results from the presented experimental work indi-
cated that various BMSW fractions have a vast po-
tential for the production of sugars that eventually can
be used for producing bio-ethanol. Dilute acid hydro-
lysis followed by steam treatment is the most efficient
pre-hydrolysis method on the selected waste among all
the pre-hydrolysis methods used in this study. High
glucose yields were obtained from both, single sub-
strates (70–89%) and multi-substrates (41–52%).
Newspaper produced the lowest glucose yield (14%)
following 96 h enzymatic hydrolysis. The contribution
of enzyme loading and acid concentration was signifi-
cantly higher (49.39 and 47.70%, respectively), than
the contribution of temperature during steam treat-
ment (0.13%) to the glucose yield.
Conversion of MSW to bio-ethanol can be an
alternative sustainable approach to disposal of waste
and reduction of the biodegradable fraction of MSW to
landfill. Technology that has been applied over the past
years to other lignocellulosic materials may be applied
to BMSW-to-bio-ethanol production; however, further
laboratory studies are needed to optimize the biocon-
version of BMSW-to-bio-ethanol production.

Fig. 5 Factorial experimental (a) Factor Effect % Contribution

design applied to a single A: acid concentration 13.12 47.70
BMSW fraction, i.e., carrot B: temperature 0.04 0.13
peelings. Pre-hydrolysis C: enzymatic loading 13.58 49.39
treatment with H2SO4 (1 and D: acid concentration and temperature 0.76 2.78
4%) and steam treatment
(121 and 134C, 15 min) and 70.57 61.66
enzymatic hydrolysis with T. (b) (c)
viride (10 and 60 FPU g–1). a 80
Predicted values , %

Factor effects on glucose

58.62 49.72
yield, b cube graph of glucose 70
yield, c comparison of actual B+
value and predicted value 60
72.80 58.22
50
C+
R2 = 0.90
40
60.85 46.27 40 50 60 70 80
B- C-
A- A+ Actual values, %

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Acknowledgments This research was undertaken at the
Department of Civil and Environmental Engineering, Labora-
tory of Environmental Engineering, Chadwick Building, Uni-
versity College London, UK, AL acknowledges the financial
support provided by the Dorothy Hodgkin Postgraduate Award,
and the technical support received at Wolfson Institute and
Department of Biochemical Engineering at the University Col-
lege London.
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