Research Article

Received: 4 February 2011 Revised: 17 June 2011 Accepted: 27 June 2011 Published online in Wiley Online Library: 5 August 2011

(wileyonlinelibrary.com) DOI 10.1002/jctb.2707

Methane production from citrus wastes:

process development and cost estimation
Gergely Forgács,a,b∗ Mohammad Pourbafrani,c,d Claes Niklasson,b
Mohammad J. Taherzadeha and Ilona Sárvári Hovátha

Abstract
BACKGROUND: Because of its extreme toxicity for microorganisms, the limonene content of citrus wastes (CWs) has been a
major obstacle to the conversion of CWs to biofuels. The main objective of this study was to develop a new process for the
utilization of CWs that can be economically feasible when the supply of CW is low.

RESULTS: Steam explosion pre-treatment was applied to improve the anaerobic digestibility of CWs, resulting in a decrease
of initial limonene concentration by 94.3%. A methane potential of 0.537 ± 0.001 m3 kg−1 VS (volatile solids) was obtained
during the following batch digestion of treated CWs, corresponding to an increase of 426% compared with that of the untreated
samples. Long-term effects of the treatment were further investigated by a semi-continuous co-digestion process. A methane
production of 0.555 ± 0.0159 m3 CH4 kg−1 VS day−1 was achieved when treated CWs (corresponding to 30% of the VS load)
were co-digested with municipal solid waste.

CONCLUSION: The process developed can easily be applied to an existing biogas plant. The equipment cost for this process is
estimated to be one million USD when utilizing 10 000 tons CWs year−1 . 8.4 L limonene and 107.4 m3 methane can be produced
per ton of fresh citrus wastes in this manner.

c 2011 Society of Chemical Industry

Keywords: citrus waste; pre-treatment; methane; limonene; cost estimation

INTRODUCTION CW as substrates for biogas production. The long-term effects of

Worldwide, citrus fruit production is about 80 million tons per year.
Almost 33% of the crops, including oranges, lemons, grapefruit,
and mandarins, are industrially processed for juice production,
where about half of the processed citrus, including peels, segment
membrane and seeds end up as wastes.1 Today, parts of these
wastes are mostly dried and marketed as either low-protein cattle
feed or used for pectin production. However, it is not economically
favorable to dry the residue owing to its high moisture content.
Therefore, the remaining part of the waste is still deposited, leading
to economical and environmental disadvantages, including but
not limited to, high transportation costs and high organic content
at the disposal sites.2
Citrus wastes (CW) consist of various soluble and non-soluble
carbohydrate polymers, which makes this waste material an ideal
feedstock for biological conversion to biofuels such as ethanol
and biogas.3,4 A process of utilizing CWs for the production of
ethanol, biogas, and limonene has recently been developed.4 The
profitability of this process is related to the supply of CW, and
economic analysis showed that 200 000 tons year−1 or more of
CW is needed to make this process economically feasible.5
The main objective of this study was therefore to develop a
simpler, more feasible process for the utilization of CW, which can
be economically feasible even when CW supply is low. CW was
treated by steam explosion treatment prior to anaerobic digestion
to eliminate the limonene content. The effect of the pre-treatment
250
the treatment were also investigated in semi-continuous anaerobic
digestion systems. In order to avoid nutrient limitation, the CW
was utilized in co-digestion with municipal solid waste (MSW),
which is the substrate normally used in a large-scale biogas plant
operating at thermophilic conditions in Borås, Sweden. Based on
the experimental results, an industrial process configuration was
proposed, and the equipment and operating costs for this process
were estimated.
MATERIALS AND METHODS
Raw materials
MSW and the bacterial inoculums used in this study were obtained
from a large-scale thermophilic biogas plant (Borås Energi & Miljö
AB, Borås, Sweden). The CW was provided by Brämhults Juice AB
∗
Correspondence to: Gergely Forgács, School of Engineering, University of Borås,
501 90, Borås, Sweden. E-mail: gergely.forgacs@hb.se
a School of Engineering, University of Borås, 501 90, Borås, Sweden
b Department of Chemical and Biological Engineering, Chalmers University of
Technology, 412 96 Göteborg, Sweden
c Process Engineering Department, Nokan Process Company, 35111 Karaj, Iran
d Present address: Department of Chemical Engineering and Applied Chemistry,

was evaluated by batch digestion tests using treated and untreated University of Toronto, 200 College Street, Toronto, ON M5S 3E5 Canada

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c 2011 Society of Chemical Industry
Methane production from citrus wastes
(Borås, Sweden). The dry matter content of CW was 20% and the
composition, in percentage of dry matter basis, was determined as
reported previously by Pourbafrani et al.4 and summarized herein
as weight/weight percentage (% w/w): glucose 8.1; fructose 12.0;
sucrose 2.8; pectin 25.0; protein 6.1; cellulose 22.0; hemicellulose
11.1; lignin 2.2; and limonene 3.8.
MSW was homogenized and enriched in biodegradable organics
by sorting away large pieces of plastic and glass in multiple steps at
the biogas plant. Additionally, the MSW was further treated in the
laboratory by shredding into smaller particles (less than 2 mm) in
a homogenizer (Wiggenhauser D500, Germany) and then stored
in plastic bags equipped with zip lock at −20 ◦ C until further use.
Treatment by steam explosion
The treatment of CWs was carried out in a 10 L high-pressure
reactor (Process & Industriteknik AB, Sweden). Before treatment,
samples of 2 kg slurry with 6.6% total solid content were prepared
by the addition of distilled water to the CWs. The slurry was
then treated by steam at a temperature of 150 ◦ C for 20 min. The
reactor was heated by direct injection of 60 bar pressure steam,
provided by a power plant located in Borås, Sweden. The treated
CW slurry was then explosively discharged into an expansion tank,
at atmospheric pressure for cool down, and used as substrate for
batch and semi-continuous digestion.
Batch digestion experiments
The batch experiments were carried out according to the method
described by Hansen et al6 using both steam treated and untreated
CWs as substrate. All of the experiments were carried out in
triplicates using 2 L glass bottles with a reaction volume of 600 mL
at thermophilic condition (55 ◦ C). Each bottle was fed with an
appropriate amount of substrate corresponding to a final volatile
solids (VS) concentration of 0.75% and subsequent addition of
distilled water up to a volume of 200 mL. Finally, 400 mL inoculum
was added to each bottle and then the reactors were sealed
with rubber septum and aluminum caps, and flushed with a gas
containing 80% N2 and 20% CO2 to ensure anaerobic conditions.
Blanks containing water replacing the substrate were also used
to determine the methane production of the inoculums itself.
Throughout the experimental periods, the bottles were incubated
at 55 ◦ C, and were shaken and moved around in the incubator
once a day.
Semi-continuous experiments
Three 5 L continuously stirred reactors, operated in a water bath at
55 ◦ C, were used in this series of experiments. One of the reactors
(R1) was considered as a control reactor and was only fed with
MSW. The second reactor (R2) was fed with a mixture of 30%
steam-treated CW and 70% MSW (corresponding to VS loading). In
the feed of the third reactor (R3), 30% of the VS loading consisted
of untreated CW and the rest was MSW. The reactors were fed
once a day, and the hydraulic retention time (HRT) was adjusted
to 21 days. The organic loading rate (OLR) was gradually increased
from 1 g VS L−1 day−1 to a final level of 3 g VS L−1 day−1 during
an initial start-up period of 15 days. The produced volume and the
composition of biogas, as well as the pH, were measured every day.
The concentration of volatile fatty acids (VFA), alkalinity (both total
alkalinity (TA) and bicarbonate alkalinity (BA)), total solids (TS), and
volatile solids (VS) in the reactor samples were determined once a
week during the operation period.
J Chem Technol Biotechnol 2012; 87: 250–255

c 2011 Society of Chemical Industry
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Analytical methods
To determine the limonene concentration remaining after the
steam explosion treatment, the steam-treated CWs were subjected
to additional enzymatic hydrolysis using a mixture of cellulase,
pectinase, and β-glucosidase enzymes.3 The concentration of
limonene was then determined by the addition of n-heptane (99%
purity), with a ratio of 1/5, to these hydrolyzate samples followed by
centrifugation at 3500×g for 30 min to extract the oil. The resulting
supernatant was then analyzed by gas chromatography–mass
spectrometry (GC-MS, Hewlett Packard G1800C, Agilent, Palo Alto,
CA), in which the carrier gas was helium. The temperature was
initially 50 ◦ C and then gradually increased to 250 ◦ C at the rate of
15 ◦ C min−1 and maintained at this final temperature for 3 min.3
Gas samples from the digesting reactors and bottles were taken
using a 0.25 mL glass syringe (VICI, Precision Sampling, Inc., USA)
equipped with pressure lock. The methane and carbondioxide
content were then analyzed directly by a gas chromatograph
(Perkin Elmer AutoSystem, USA) equipped with a packed column
(Perkin Elmer, 6 × 1.8 OD, 80/100, Mesh, USA) and a thermal
conductivity detector (Perkin Elmer, USA) with inject temperature
150 ◦ C. The carrier gas was nitrogen operated with a flow rate of
20 mL min−1 at 60 ◦ C. Data treatment was carried out according
to Hansen et al.6 All the results of methane volumes are presented
under normal conditions (273 K, atmospheric pressure).
The concentrations of volatile fatty acids were determined by
high performance liquid chromatography (HPLC, Waters 2695,
Millipore, Milford, USA) equipped with a refractive index (RI)
detector (Waters 2414) and an ion-exchange column (Aminex
HPX-87H column, Bio-Rad, USA), at 60 ◦ C using 5 mmol L−1
H2 SO4 as eluent with a flow rate of 0.6 mL min−1 . For alkalinity
determination, a double end point potentiometric titration was
used. Bicarbonate alkalinity (BA) and total alkalinity (TA) were
measured by titration with 0.05 mol L−1 HCl to the end point of
5.75 and 4.00, respectively, according to a method described by
Larsson.7
Cost estimations
The steam explosion treatment process was designed to treat
10 000 tons of CW per year. Details on equipment material and
sizing can be found in a previous publication.5 The costs of the
equipment (sum of purchasing and installing equipment) were
calculated using the software Icarus Process Evaluator (IPE) (Aspen
0.6
0.5
CH4 (m3/kg VS)
0.4
0.3
0.2
0.1
0.0
0 10 20 30 40
Time (days)
MSW
Steam explosion treated CW
Untreated CW
Figure 1. Accumulated methane production expressed in m3 CH4 kg−1
VSadded , obtained during thermophilic batch digestion of MSW (•), as well
as steam explosion treated (◦) vs. untreated (
) CW samples.
251
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 www.soci.org G Forgács et al.

Table 1. Summary of process parameters, i.e. TS, VS, total alkalinity, and total VFA, as well as individual VFA concentrations, during anaerobic
semi-continuous digestion of MSW alone (R1), and co-digestion of MSW mixed with treated (R2) and untreated CW (R3), respectively

Day – 15 Day 1 Day 11 Day 22 Day 35 Day 49

R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3 R1 R2 R3

TS (%) 2,40 2,40 2,40 2,55 2,60 2,45 2,37 2,47 2,39 2,22 2,33 2,71 2,17 2,28 – 2,25 2,26 –
VS (%) 1,60 1,60 1,60 1,80 1,82 1,65 1,76 1,55 1,70 1,70 1,51 – 1,65 1,62 – 1,72 1,66 –
Alkalinity (mg L−1 ) 8650 9017 9017 8100 8540 8340 8450 8306 8306 8726 8560 8560 7256 7324 – 7344 7266 –
Acetic acid 0,35 0,35 0,35 1,02 1,33 0,92 0,42 0,47 1,88 0,38 0,30 1,72 0,42 0,43 – 0,30 0,35 –
Propionic acid 0,51 0,51 0,51 0,60 0,46 1,62 0,54 0,59 2,31 0,25 0,61 2,03 0,28 0,51 – 0,43 0,42 –
Iso-butyrate 0,18 0,18 0,18 0,12 0,15 0,22 0,15 0,15 0,32 0,04 0 0,25 0,10 0,20 – 0 0 –
Butyrate 0,29 0,29 0,29 0,18 0,25 0,10 0,10 0,10 0,09 0 0 0 0 0 – 0 0 –
Iso-valerate 0,20 0,20 0,20 0,12 0,20 0,32 0,20 0,24 0,48 0,07 0,30 0,43 0,33 0,32 – 0,30 0,29 –
Valerate 0,42 0,42 0,42 0,22 0,12 0,22 0,22 0,32 0,21 0,11 0 0,13 0 0,31 – 0,20 0 –
Capriorate 0 0 0 0 0 0 0 0 0,44 0 0 1,94 0 0 – 0 0 –
Total VFA (g L−1 ) 1,95 1,95 1,95 2,26 2,51 3,40 1,63 1,87 5,72 0,84 1,21 6,51 1,13 1,77 – 1,23 1,06 –

Tech, USA).5 The effect of different CW capacities on the equipment
cost for these process steps was investigated.
RESULTS
Batch experiments
In the batch anaerobic digestion series, the methane production of
pre-treated CW was compared with that of untreated samples. The
results are presented in Fig. 1. The accumulative methane produc-
tion increased to 0.537 ± 0.001 m3 kg−1 VSadded , corresponding to
an increase of 426% compared with 0.102±0.021 m3 kg−1 VSadded
for the untreated samples. The anaerobic digestion of MSW, alone,
resulted in 0.557 ± 0.026 m3 CH4 kg−1 VSadded .
Semi-continuous experiments
The results of semi-continuous experiments are summarized in
Table 1 and in Fig. 2. Reactor R1 was considered the control reactor
and was operated with MSW only. In R2 and R3, treated or untreated
CWs, respectively, were co-digested with MSW. After a 15 day
startup period, the specific production of methane remained
between 0.5 m3 kg−1 VS day−1 and 0.6 m3 kg−1 VS day−1 during a
following stable operating period of 20 days (Fig. 2(A)). However,
in R3 a sharp decrease in methane production was observed after
20 days (Fig. 2(A)). Moreover, the pH remained around 7.5 until
the end of the experiment in the control reactor and in R2, while
pH started to decrease after 10 days of stable operation to a final
value of 5.5 in R3 (Fig. 2(B)). The TS and VS values remained stable
during the operating period (i.e. 50 days) in the first two reactors
(Table 1). Although TA decreased somewhat during operation in
these two reactors, the total VFA versus TA ratio always remained
below 0.3, indicating no disturbance in the process (Table 1). In
contrast, as is shown in Table 1 and in Fig. 2(C), the total VFA
concentration increased from 1.95 g L−1 to 6.51 g L−1 in R3. In
addition, the degradation rate decreased in R3 as indicated by the
increasing TS values (from 2,40% to 2.71%) at the outlet (Table 1).
Cost estimations
A typical block flow diagram detailing the process of steam
explosion pre-treatment and limonene recovery presented in this
study is shown in Fig. 3(A). The CW is transferred to the reactors
252
water and hydrolyzed at 150 ◦ C for 20 min without adding any
extra chemical. More than 94% of the limonene content of CW is
released during the hydrolysis process and flashed in the expansion
tank. The vapour outlet of the expansion tank is condensed and its
limonene content decanted in a decanter. This process can easily
be implemented in an existing biogas plant, by pumping the
hydrolyzate to the digester and utilizing it for methane production
within a co-digestion process, resulting in the production of 8.4 L
limonene and 107.4 m3 methane per ton of wet CW.
The cost of the equipment was estimated to be about 1 million
USD, for a capacity of 10 000 tons CW year−1 . This cost includes
installation and purchase cost of equipment for the steam
explosion pre-treatment step as well as for the limonene recovery.
The price of land is, however, excluded. The difference in the cost
of equipment with the available quantity of CWs is presented in
Fig. 3(B).
The operating cost of the process is the sum of transportation
and handling cost for CW, utility, maintenance, insurance and
labour costs (Table 2). The utility requirements of the process
including steam, cooling water and electricity are 1100, 700, 5 kW
per ton of dried CW, respectively. The required steam can be
provided by burning 22% of the methane produced in a steam
boiler. One labour can operate the process in 8 h working time.
The transportation and handling costs depend on the distance
between the treatment plant and the juice factory,5 and here it is
assumed to be 10 USD ton−1 wet CW. Changes of operating costs
versus capacity of plant are presented in Fig. 3(C).
DISCUSSION
In this study, the utilization of citrus peel waste for methane
production was examined. Earlier investigations showed that
factors such as the presence of limonene in CW and other
aromatic acids present in the peel oil have negative effects on
the digestion process.8,9 The batch digestion assay in this study
showed that untreated CW has a low methane potential of
0.102 m3 kg−1 VSadded . In comparison, Kaparaju et al.10 reported
a methane production potential from untreated industrial orange
waste of 0.49 m3 kg−1 VSadded . However, the organic loading
was lower than that in our study and, moreover, the system
was buffered by the addition of NaHCO3 to keep the pH at an

using a conveyor and then mixed with a specified volume of appropriate level for anaerobic digestion.

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c 2011 Society of Chemical Industry J Chem Technol Biotechnol 2012; 87: 250–255
Methane production from citrus wastes www.soci.org

A
0.7
Start-up period
0.6

CH4 (m3/kg VS)

0.5
0.4
0.3
0.2
0.1
0.0
-20 -10 1 10 20 30 40 50 60
Time (days)
B 8.0

7.5

7.0
pH

6.5
Start-up period
6.0

5.5

5.0
-20 -10 1 10 20 30 40 50 60
Time (days)
C 7
Start-up period
6
5
VFA ( g/L)

4
3
2
1
0
-20 -10 1 10 20 30 40 50 60
Time (days)

Control reactor with 100% MSW

70% MSW +30% steam exploded CW
70% MSW +30% untreated CW

Figure 2. Semi-continuous anaerobic digestion of MSW as a control (•), as well as co-digestion of MSW mixed with treated CW (◦) and untreated CW
(
), respectively (the minus values of days corresponding to the start-up period). The changes in CH4 production (A), pH (B), and total volatile fatty acids
(C) are presented at organic loading (OLR) of 3 gVS L−1 day−1 , hydraulic retention time (HRT) 21 days, and thermophilic conditions at 55 ◦ C.

Mizuki et al.8 found a limiting limonene load of 65 µL L−1 in
a semi-continuous system with 10 days retention time. However,
although the limonene loading rate was lower than 65 µL L−1 per
day in our study, strong inhibition was observed in R3 (Fig. 2),
which can be explained by the higher sensitivity of a thermophilic
system compared with the mesophilic one.11
Previously, Martı́n et al.9 observed 0.33 m3 kg−1 VSadded
methane yield from orange peel, after removing 70% of
its D-limonene content by steam distillation pre-treatment.
In contrast, when CWs were pre-treated by steam explosion
in this study, methane production increased to a value of
0.537 ± 0.001 m3 kg−1 VSadded during the following anaerobic
digestion, an increase of 426% over that for the untreated CWs
(Fig. 1). Moreover, the D-limonene concentration in the CW hy-
drolyzate decreased by 94.3% after treatment, resulting in a
concentration of only 0.01 w/w% remaining. An additional advan-
tage with this pre-treatment is that the released limonene, which
is removed through flashing of the hydrolyzate into the expansion
sion treatment opens up the structure of complex polymers in
the raw material resulting in a partial hydrolysis of carbohydrates,
which further improves the biodegradability during the following
anaerobic digestion, leading to improved methane production.
When treated CW was co-digested with MSW (R2), a methane
production of 0.555 ± 0.016 m3 kg−1 VS day−1 was achieved.
During the operating period, the total VFA concentration remained
under 2 g L−1 and in addition, the pH, TS, and VS values
remained stable (Table 1 and Fig. 2). In comparison, when the
semi-continuous system was operating with feed containing
untreated CW, the process failed after approximately 26 days
(R3 in Table 1 and Fig. 2). Because of the accumulation of peel
oils and the related inhibition effects, the total VFA content
increased to 6.51 g L−1 (Fig. 2(C)). The accumulation of VFAs has
been shown to be a phenomenon typical of stressed digesters.12
Moreover, the accumulation of VFAs results in a decrease in pH
(R3 in Fig. 2(B) and (C)). The optimal pH for anaerobic digestion
is between 7 and 8.13,14 If the pH in the reactors drops below pH
253

tank (Fig. 3(A)), can be recovered.5 Furthermore, the steam explo- 6.2, methanogenesis will be inhibited and as a consequence, the

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 www.soci.org G Forgács et al.

A Citrus
Waste Water Steam
Condenser
Expansion Tank Decanter

Limonene

Water
Conveyor

Reactor
Valve
To Biogas
Plant

B 3.5 C 800
Equipment cost (million USD)

Operating cost (1000 USD/year)

3.0 700

600
2.5
500
2.0
400
1.5
300

1.0 200

0.5 100
0 10000 20000 30000 40000 50000 60000 70000 0 10000 20000 30000 40000 50000 60000 70000
CW capacity (ton/year) CW capacity (ton/year)

Figure 3. Block flow diagram (A) of the steam explosion pre-treatment process combined with limonene recovery. The prediction of equipment (B) and
operating (C) cost for the process as a function of citrus waste capacity.

Table 2. Cost elements used in the calculation of operating cost

ACKNOWLEDGEMENT
The authors are grateful to the ‘Sjuhärads Kommunalförbund’
Price Unit for financial support for this work. The authors thank Tatiana
Svedén, Solmaz Aslanzadeh, and Peyruz Özmen for experimental,
Utilities
technical, and analytical support. Dr Dag Henriksson is gratefully
Electricity 50 USD per MWh
thanked for his advice and useful comments.
Cooling water 0.3 USD per ton
Other costs
Labour 70 000 USD per employee year REFERENCES
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