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Bioresource Technology 101 (2010) 75237528

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

Enhanced solid-state anaerobic digestion of corn stover by alkaline pretreatment


Jiying Zhu 1, Caixia Wan 1, Yebo Li *
Department of Food, Agricultural, and Biological Engineering, The Ohio State University/Ohio Agricultural Research and Development Center, 1680 Madison Ave.,
Wooster, OH 44691-4096, USA

a r t i c l e

i n f o

Article history:
Received 10 February 2010
Received in revised form 19 April 2010
Accepted 24 April 2010
Available online 21 May 2010
Keywords:
Anaerobic digestion
Solid-state fermentation
Corn stover
Alkaline pretreatment
Biogasication

a b s t r a c t
Alkaline pretreatment was applied to enhance biogas production from corn stover through solid-state
anaerobic digestion. Different NaOH loadings (1%, 2.5%, 5.0% and 7.5% (w/w)) were tested for solid-state
pretreatment of corn stover. Lignin degradation during pretreatment increased from 9.1% to 46.2% when
NaOH concentration increased from 1.0% to 7.5%. The NaOH-pretreated corn stover was digested using
efuent of liquid anaerobic digestion as inoculum and nitrogen source. NaOH loading of 1% did not cause
signicant improvement on biogas yield. The highest biogas yield of 372.4 L/kg VS was obtained with 5%
NaOH-pretreated corn stover, which was 37.0% higher than that of the untreated corn stover. However, a
higher NaOH loading of 7.5% caused faster production of volatile fatty acids during the hydrolysis and acidogenesis stages, which inhibited the methanogenesis. Simultaneous NaOH treatment and anaerobic
digestion did not signicantly improve the biogas production (P > 0.05).
2010 Elsevier Ltd. All rights reserved.

1. Introduction
Lignocellulosic biomass, such as crop residues and forest waste,
is mainly composed of cellulose, hemicelluloses and lignin. It has
been well studied for ethanol production in recent years. The complex structure of native lignocellulosic biomass creates recalcitrance to enzymatic hydrolysis by cellulolytic microbes during
anaerobic digestion. In addition, due to the low cellulolytic activity
and low specic growth rate of cellulolytic microbes in anaerobic
digestion, hydrolysis of native lignocellulosic biomass is usually
slow (Noike et al., 1985). As a consequence, hydrolysis appears to
be the most rate-limiting stage among the four stages (hydrolysis,
acidogenesis, acetogenesis, and methanogenesis) of anaerobic
digestion (Lu et al., 2007). Pretreatment of lignocellulosic biomass
may represent a feasible solution to improve digestion efciency
and biogas production.
Alkaline pretreatment is one of the current leading pretreatment methods as it has shown several positive effects, including
solubilization of lignin and neutralization of various acidic products degraded from the lignocellulosic complex (Hendriks and Zeeman, 2009; Pavlostathis and Gossett, 1985). Additionally, the
presence of a small amount of residual alkali in the treated solids
may be helpful for preventing the drop of pH during subsequent
acidogenesis process (Pavlostathis and Gossett, 1985). Therefore,
alkaline pretreatment is more effective and compatible with subse* Corresponding author. Tel.: +1 330 263 3855; fax: +1 330 263 3670.
E-mail address: li.851@osu.edu (Y. Li).
1
These authors contributed equally to this work.
0960-8524/$ - see front matter 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2010.04.060

quent anaerobic digestion when compared to other pretreatment


methods such as thermochemical pretreatment. Among the three
kinds of alkali (NaOH, KOH, lime) tested for pretreatment of rice
straw, NaOH was found to be most effective for lignin removal
and biogas production (Yang et al., 2009). Zheng et al. (2009) reported that a 72.9% increase in biogas production was obtained
from corn stover pretreated with 2% NaOH for 3 days. Although
alkaline pretreatment, especially with NaOH, can lead to signicant
improvement on biogas production, toxicity of cations on methanogens was also reported (Feijoo et al., 1995). Compared to the
conventional alkaline pretreatment, solid-state alkaline pretreatment only uses a limited amount of water to saturate the feedstock
and there is no wastewater generated. Solid-state alkaline pretreatment of crop residues has been shown to be effective for
improving biodegradability and biogas production. Pang et al.
(2008) reported that corn stover pretreated with 6% NaOH at 80%
moisture content and ambient temperature (20 2 C) for 3 weeks
resulted in 48.5% more biogas yield over the untreated.
Solid-state anaerobic digestion (SS-AD) is generally used to process organic waste with high solid content (2035%). It has been
successfully implemented in Europe for processing the organic
fraction of municipal solid waste (OF-MSW) (Bolzonella et al.,
2003; Mata-Alvarez et al., 2000). Compared to liquid phase anaerobic digestion (0.515% solid loading), SS-AD is relatively stable
and requires less energy input. However, the disadvantages of
SS-AD include the requirements of large amounts of inoculum
(up to 50%), much longer retention time (three times longer than
liquid AD), and nitrogen supplementation when lignocellulosic
biomass is used. In this study, an integrated anaerobic digestion

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J. Zhu et al. / Bioresource Technology 101 (2010) 75237528

system was tested to evaluate the effectiveness of biogas production from lignocellulosic biomass in SS-AD using efuent of liquid
AD as inoculums and nitrogen source. This process can overcome
the major disadvantages for liquid anaerobic digestion (treatment
of efuent) and conventional SS-AD (large volume recycling of digestate and/or leachate for inoculation and costly nitrogen source
supplementation). It can also potentially reduce the production
cost and increase the energy efciency of biogas production. The
main objective of this study was to investigate the effect of NaOH
pretreatment on the biogas production from corn stover via SS-AD.
Lignin and holocellulose (cellulose and hemicellulose) degradation
of corn stover during NaOH pretreatment were studied. The reduction of total solids (TS) and volatile solids (VS) of NaOH-pretreated
corn stover and the variation of volatile fatty acids, pH and alkalinity during digestion were also investigated.

at the end of the digestion tests. To analyze the pH, alkalinity and
VFA during digestion, nine identical reactors with 5% NaOH-pretreated corn stover were set up at the same time and terminated
on day 3, 6, 9, 12, 15, 18, 25, 32 and 40 for sampling and subsequent analysis.

2. Methods

The cellulose, hemicellulose, and lignin contents of the untreated and NaOH-pretreated corn stover samples were determined following NREL Laboratory Analytical Procedure (Sluiter
et al., 2008). A two-stage acid hydrolysis was used for fractionating
the corn stover samples. HPLC (Agilent 1200 series, MN, USA)
equipped with a Biorad Aminex HPX-87P column and a refractive
index detector (RID) was used for analysis of monomeric sugars
in the acid hydrolysate. The temperatures of the column and the
RID were maintained at 80 and 55 C, respectively. Water was used
as the mobile phase with a ow rate of 0.6 ml/min. Cellulose and
hemicellulose contents were calculated from the corresponding
monomers. The acid insoluble lignin was measured by gravimetric
analysis and the acid soluble lignin was measured by UVvis spectroscopy (Biomate3, Thermo-scientic, MA, USA). Lignin, cellulose and hemicellulose degradation was dened as the
percentage of lignin, cellulose and hemicellulose reduction,
respectively.
TS, VS, pH and alkalinity were measured according to the Standard Methods for the Examination of Water and Wastewater
(APHA, 2005). Total carbon and total nitrogen were determined
with an elemental analyzer (Elementar Vario Max CNS, Elementar
Americans, Mt. Laurel, NJ, USA). The samples for pH measurement
were prepared by suspending 5 g wet digestate into 50 ml distilled
water. The samples for VFA measurement were prepared by suspending 10 g wet digestate in 30 ml distilled water followed by
centrifugation (8000 rpm, 5 min). One fraction of supernatant
was acidied with three fractions of formic acid and then ltered
through 0.2 lm nylon syringe lter for VFA analysis. VFA analysis
was conducted using a gas chromatograph (GC) (HP5890, Agilent
Technologies, Wilmington, DE, USA) equipped with a ame ionization detector (FID) and a 30 m  0.32 mm  0.5 lm Stabilwax-DA
Fused Silica column. The temperatures of the injection port and
detector were 230 and 230 C, respectively. Helium was used as
a carrier gas with a ow rate of 20 ml/min. The daily biogas production was measured using a wet drum gas meter (TG 5, Calibrated Instruments Company, NY, USA). Gas composition was
determined using a GC (HP6890, Agilent Technologies, Wilmington, DE) equipped with a 30 m  0.53 mm  10 lm alumina/KCl
deactivation column and a thermal conductivity detector (TCD).
Helium was used as the carrier gas at a ow rate of 5.2 ml/min.
The temperatures of the injector and detector were 150 and
200 C, respectively. The biogas yield was expressed as the volume
of biogas produced based on the initial total VS in corn stover. The
alkalinity and VFA concentration expressed in were based on the
digestate (wet basis).

2.1. Feedstock and inoculum


Corn stover was collected from the farm of Ohio Agricultural Research and Development Center (OARDC) in Wooster, OH and airdried to a moisture content less than 10%. The dried corn stover
was than ground to pass through a 5 mm sieve and stored in airtight containers prior to use. Efuent of liquid anaerobic digestion
was supplied by Akron facility of quasar energy group and used as
inoculum. The characteristics of corn stover and efuent are presented in Table 1.
2.2. Solid-state NaOH pretreatment
Two hundred grams of corn stover (dry basis) were mixed
evenly with 200 ml NaOH solution (0.25, 0.625, 1.25, and
1.85 M). The corresponding NaOH loadings over the substrate solids were 1.0%, 2.5%, 5.0% and 7.5% (w/w), respectively. The NaOH
soaked corn stover was kept at ambient temperature (20 0.5 C)
in sealed zipper plastic bags for 24 h. Samples were taken for compositional analysis before the anaerobic digestion tests.
2.3. Solid-state anaerobic digestion
The pretreated corn stover was mixed with 800 g efuent to obtain a C/N ratio of 18 and a TS content of 22% which are optimal for
methane production (Zhu et al., 2009). The inoculated corn stover
was placed into 2 L reactors and digested at 37 C for 40 days. Biogas was collected daily with 5 L Tedlar gas bags throughout the
digestion process for gas composition and production analysis.
TS, VS, pH, alkalinity, and volatile fatty acids (VFA) were analyzed

Table 1
Characteristics of corn stover and efuent.
Parameter

Corn stover

Efuent

Total solids (%)


Volatile solids (%)
Total carbon (%)
Total nitrogen (%)
C/N ratio
pH value
VFA (g/kg)
Alkalinity (g CaCO3/kg)
Cellulose (%)
Hemi-cellulose (%)
Acid-insoluble lignin (%)
Acid-soluble lignin (%)
Lignin (%)

94.7 0.3
88.0 0.8
43.6 0.4
0.6 0.0
67.6 0.0
ND
ND
ND
40.7 1.6
22.5 0.3
20.2 0.5
1.5 0.3
21.7 0.5

9.8 0.1
6.4 0.1
4.1 0.2
0.7 0.0
5.8 0.3
8.3 0.1
6.8 0.2
19.9 0.1
ND
ND
ND
ND
ND

ND = not determined.

2.4. Concurrent NaOH pretreatment and anaerobic digestion


Two hundred grams of corn stover (dry basis) were mixed
evenly with 200 ml NaOH solution (0.25, 0.625, and 1.25 M) and
800 g efuent. The mixture was immediately fed into the SS-AD
reactor for digestion tests. The digestion conditions and sampling
procedures were the same as those described in Section 2.3.
2.5. Analytical methods

2.6. Statistical analysis


All the NaOH pretreatment and digestion tests were conducted
in duplicates and the average values were reported. The software

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J. Zhu et al. / Bioresource Technology 101 (2010) 75237528

3.1. Degradation of corn stover during pretreatment


NaOH pretreatment of corn stover for 24 h caused signicant
degradation of corn stover (Table 2). An increase in NaOH concentration resulted in increased lignin removal. When the NaOH concentration increased from 1% to 7.5%, the lignin degradation
increased from 9.1% to 46.2%, respectively. The highest hemicellulose loss of 13.8% was obtained with 7.5% NaOH addition, which
was about two times of that obtained with 1.05.0% NaOH addition. The cellulose loss caused by pretreatment was less than 7%
for all testes. The low cellulose degradation was partially attributed to the physical protection from lignin and hemicellulose.
Alkaline treatment generally caused the swelling of cellulose bers
and modication of cellulose crystallinity rather than direct degradation (Hendriks and Zeeman, 2009). Meanwhile, the hetero-structures of hemicellulose branched with short lateral chains
(Hendriks and Zeeman, 2009) contributed to the higher hemicellulose degradation at a higher NaOH concentration. Our results indicated that NaOH pretreatment can preserve most of the
carbohydrates but caused substantial lignin degradation during
24-h pretreatment at room temperature. In addition to signicant
lignin degradation by alkaline pretreatment, an increase in water
extractives was also reported (Pang et al., 2008; Zheng et al.,
2009). Therefore, substantial lignin degradation, increased substrate porosity, and exposed remaining carbohydrate make the
NaOH-pretreated corn stover more accessible to cellulolytic microorganism than the untreated.

25
untreated

Daily biogas yield (L/kg VS)

3. Results and discussion

1.0% NaOH
20

2.5% NaOH
5.0% NaOH
7.5% NaOH

15

10

0
0

12

24

28

32

24

28

32

36

40

untreated
1.0% NaOH
2.5% NaOH
5.0% NaOH
7.5% NaOH

350
300
250
200
150
100
50
0
0

12

16

20

36

40

Time (d)

Table 2
Effect of 24-h alkaline pretreatment on corn stover degradation.
Degradation (%)
Cellulose

Hemicellulose

Lignin

6.5 1.4
5.8 0.4
5.1 0.8
6.8 0.1

6.1 1.4
4.0 0.5
6.9 1.1
13.8 3.1

9.1 0.4
11.4 0.3
31.4 0.6
46.2 0.7

80
70

Methane content (%)

3.2.1. Biogas production


NaOH-pretreated corn stover was directly digested for biogas
production without any washing or detoxication. As shown in
Fig. 1a, the daily biogas production of 1.0% and 2.5% NaOH-pretreated corn stover reached 4.6 and 4.1 L/kg VS, respectively, during 24 h of digestion, which is similar to that of the untreated
corn stover. The rapid initial biogas production is due to readily
biodegradable organic matter in the substrate irrespective of the
pretreatment. A temporary decline after the rst day might be
caused by dissipation of substrate readily available for microbial
decomposition (Ahn et al., 2009). The daily biogas yield of 1.0%
and 2.5% NaOH-pretreated corn stover reached their peak values
of 18.2 and 22.5 L/kg VS, respectively, on day 7. Thereafter, the daily biogas yield decreased rapidly to 8.3 and 9.4 L/kg VS on day 12
and nally decreased to 2.1 and 2.6 L/kg VS on day 40. Biogas yield
of the untreated corn stover recovered slowly and reached the peak
value (20.7 L/kg VS) on day 9. This result indicates that the 1% and
2.5% NaOH pretreated corn stover were more easily accessible to
hydrolytic bacteria at the early stage of digestion. For 5% NaOHpretreated corn stover, the daily biogas yield reached 7.1 L/kg VS
on day 3 and then decreased slightly followed by a gradual in-

1.0
2.5
5.0
7.5

20

400

3.2. Solid-state anaerobic digestion

NaOH concentration (%)

16

Time (d)

Cumulative biogas yield (L/kg VS)

SAS 9.1 (SAS Inc., Cary, NC, USA) was used for analysis of variance
(ANOVA).

60
50
40
untreated

30

1.0% NaOH
2.5% NaOH

20

5.0% NaOH

10

7.5% NaOH

0
0

12

16

20

24

28

32

36

40

Time (d)
Fig. 1. Biogas production of 24-h alkaline pretreated corn stover: (a) daily biogas
yield, (b) cumulative biogas yield, and (c) methane content of biogas.

crease. The daily biogas yield reached a peak value of 19.9 L/kg VS
on day 11 and decreased slowly to 5.9 L/kg VS on day 28. During
this period, the biogas production of 5% NaOH-pretreated corn stover was substantially higher than that of all other treatments
(Fig. 1a and b). Since 5% NaOH pretreatment removed 31.4% lignin
from corn stover, the hydrolysis may not be a rate-limiting step
during the digestion process. However, sodium ions and aromatic
compounds derived from lignin degradation are known to be
inhibitory to methanogens (Gossett et al., 1982), which might contribute to the lower daily biogas yield in the rst 10 days when
compared to 1.0% and 2.5% NaOH-pretreated corn stover (Fig. 1a).

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J. Zhu et al. / Bioresource Technology 101 (2010) 75237528

Table 3
Change of VFA, pH and alkalinity during anaerobic digestion.

a
b
c

NaOH
concentration
(%)

pHia

pHfb

Untreated
1.0
2.5
5.0
7.5c

8.5 0.0
8.5 0.0
8.6 0.0
8.9 0.1
9.5 0.4

8.5 0.0
8.3 0.0
8.5 0.0
8.5 0.1
6.0 0.3

Alkalinityi

Alkalinityf

VFAf

(g CaCO3/
kg)

(g CaCO3/
kg)

(g/kg)

13.6 0.7
14.3 0.8
17.6 0.9
21.8 0.8
26.4 1.3

11.9 0.2
14.0 0.1
15.3 0.7
20.2 1.3
10.7 0.5

2.0 0.1
2.6 0.2
2.0 0.7
2.4 0.6
22.1 1.1

i initial values at the beginning of the digestion.


f nal value after 40 days of digestion.
f nal value after 20 days of digestion.

Corn stover pretreated with 7.5% NaOH caused failure of methane production. It was observed that corn stover pretreated with
7.5% NaOH resulted in a low daily biogas yield with low methane
content and the digestion ceased on day 13 (Fig. 1a and c). A highly
acidic environment (pH 6.0) caused by the accumulation of VFA
inhibited the methanogenic bacteria when the 7.5% NaOH-pretreated corn stover was digested (Table 3). Similar results were reported by Penaud et al. (1999) who observed that methane
production decreased when 5 g/L or higher NaOH loading was
used. Although sodium ions were not found to inhibit the methane
production in the study of Penaud et al. (1999), sodium concentrations of 0.250.4 M (1016 g/L) were reported to impose sodium
ion toxicity on methanogenic bacteria by Rinzema et al. (1988).
The cumulative biogas yields of corn stover pretreated with different NaOH loadings are presented in Fig. 1b. The highest cumulative biogas yield (372.4 L/kg VS) for 40-day digestion was obtained
with 5.0% NaOH-pretreated corn stover, which was 37.0% higher
than that of the untreated samples. The cumulative biogas yields
for 40 days reached 266.8 and 275.9 L/kg VS for 1.0% and 2.5%

Table 4
Degradation of substrates after 40-day anaerobic digestion.

NaOH concentration
(%)

Degradation (%)
TS

VS

Cellulose

Hemicellulose

Untreated
1.0
2.5
5.0
7.5a

21.8 0.4
21.2 0.4
22.2 0.2
25.4 1.3
7.7 0.4

35.3 2.3
35.8 0.3
38.4 2.0
44.4 0.8
13.7 0.7

44.5 3.2
43.2 1.9
44.1 6.2
65.1 0.2
27.6 2.5

45.7 3.8
43.6 3.2
46.0 3.3
63.6 2.5
61.3 3.7

Twenty days of digestion.

NaOH-pretreated corn stover, respectively, which were not significantly different from the control (untreated corn stover). These results indicated that NaOH concentrations lower than 2.5% were not
effective in improving the biogas production. On the other hand,
7.5% NaOH-pretreated corn stover inhibited the methane
production.
Among the functional digesters, the methane contents increased gradually and reached about 60% on day 10 (Fig. 1c). There
were variations of methane content among different treatments,
but they were not signicantly different. The methane contents
ranged between 50% and 60% after day 10 for all the functional
digesters.
3.2.2. Degradation of corn stover
The efciency of solid-state anaerobic digestion was evaluated
in terms of TS and VS reduction as well as carbohydrate reduction.
As shown in Table 4, VS reduction increased as NaOH loading increased from 1.0% to 5.0%. Pretreatment with 5.0% NaOH caused
the highest VS reduction (44.4%) in SS-AD. No signicant improvement on VS reduction was observed with 1.0% and 2.5% NaOH pretreatment. The TS reduction followed a similar pattern as that of
the VS. Cellulose and hemicellulose reductions of 1.02.5%
NaOH-pretreated corn stover during SS-AD were close to that of
the untreated samples. The highest cellulose and hemicellulose
reductions of 65.1% and 63.1%, respectively, were observed with
5% NaOH-pretreated corn stover, which is in agreement with the
highest biogas production at this condition. The lowest cellulose
reduction of 27.6% was observed with 7.5% NaOH-pretreated corn
stover. The hemicellulose degradation of 7.5% NaOH-pretreated
corn stover reached 61.3% which was higher than that of 1.0%
and 2.5% NaOH-pretreated corn stover. This may be caused by
the high alkali residue in the 7.5% NaOH-pretreated corn stover
which continuously degraded the hemicellulose during the digestion process.
3.2.3. Variation of VFA, pH and alkalinity
Volatile fatty acids (mainly acetic acid, propionic acid, and butyric acid) produced during the acidogenic stages are vital to the
anaerobic digestion process. The degradation of propionate and
butyrate by syntrophic acetogenic bacteria (e.g. Syntropher wolinii,
syntrophomonas wolfei) produces acetic acid that is subsequently
degraded into methane and CO2 by acetoclastic methanogens
(Montero et al., 2008). The evolution of VFAs plays an important
role in maintaining efcient anaerobic digestion as it strongly affects the pH value, alkalinity, and the activity of methanogens
(Buyukkamaci and Filibeli, 2004; Moller et al., 2004). The irrevers-

Fig. 2. Variation of VFA, pH and alkalinity during anaerobic digestion of 5.0% NaOH-pretreated corn stover.

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J. Zhu et al. / Bioresource Technology 101 (2010) 75237528

3.3. Simultaneous alkaline treatment and SS-AD


In order to simplify the process and reduce the capital cost,
simultaneous alkali treatment and digestion (SATD) was also
tested. Compared to the 24 h-pretreated corn stover, the highest
peak daily biogas production of SATD was delayed from day 7 to
day 9 for treatments with 1.0% and 2.5% NaOH addition. The
24-h pretreated corn stover can provide readily degradable carbohydrates to hydrolytic bacteria, which enhanced the biogas in the
early stage. In SATD, corn stover needs to be depolymerized by
alkali before hydrolysis and acidogenesis, resulting in no improvement on daily biogas production by alkali addition in the rst
several days. As shown in Fig. 3b, the 40-day accumulative biogas
yield of corn stover with 1.0% NaOH addition was 278.9 L/kg VS,
which was close to that of the untreated corn stover (275.9 L/kg VS).

However, the biogas production at 2.5% NaOH loading in SATD


between days 9 and 16 was higher than that of the untreated
(Fig. 3a), which caused 9.0% increase in the cumulative biogas yield
over the untreated.
SATD with 5% NaOH addition was inhibited at the early stage,
resulting in a lower daily biogas production. The cumulative biogas
production was 199.4 L/kg VS, which was 27% lower than that
of the untreated. A high initial pH value (pH 9.42) caused by 5%
NaOH addition might be the major reason causing the inhibition

Daily biogas yield (L/kg VS)

25

untreated
1.0% NaOH

20

2.5% NaOH
5.0% NaOH
15

10

0
0

12

16

20

24

28

32

36

40

24

28

32

36

40

Time (d)

Cumulative biogas yield (L/kg VS )

400

untreated
350

1.0% NaOH
2.5% NaOH

300

5.0% NaOH
250
200
150
100
50
0
0

12

16

20

Time (d)

80
70

Methane content (%)

ible acidication of the digestion, resulting from rapid hydrolysis


and acidogenesis, is the major challenge of anaerobic digestion,
as it can cause inhibition of methanogenesis or failure of the digestion (Veeken and Hamelers, 1999; Wang et al., 2009).
Variation of total short chain VFAs during anaerobic digestion of
5% NaOH-pretreated corn stover is presented in Fig. 2. Acetic acid
was the dominant volatile fatty acids. There was no signicant
accumulation of propionic acid and butyric acid, probably because
of the sufcient propionate- and butyric-degrading syntrophs in
the inoculum which can rapidly convert propionic acid and butyric
acid to acetic acid (Montero et al., 2008). The acetic acid increased
rapidly after starting the test and reached a maximum of 21.5 g/kg
on day 12. During this period, the acetic acid production rate was
apparently higher than the acetic acid consumption rate. After 15
days, acetic acid concentration rapidly dropped to 12 g/kg and remained stable until the end of the digestion process. During this
period, the digestion reached the stabilization stage (methanogenesis) with a balance between the production and consumption of
acetic acid and a constant methane content of 5060% was
obtained.
The pH is an important parameter for proper operation and
monitoring of the anaerobic digestion process. It strongly depends
on VFA and buffering capacity. The pH value decreased with the increase in VFA production by acidogenic bacteria. The lowest pH value of 6.6 was observed on day 12, which corresponds to the
maximum VFA production of 24.7 g/kg. Subsequently, pH values
increased between 8.4 and 8.5 with a decrease in VFA concentration, indicating the further conversion of VFA to methane by methanogens. The alkalinity had a similar prole as pH, reaching the
minimum level of 17.1 g CaCO3/kg on day 12 and subsequently increased to 20.421.1 g CaCO3/kg on day 12 (Fig. 2). The increase of
alkalinity during the later stage could be mainly due to the consumption of VFA by methanogens, suggesting that the alkali
(OH ) remaining in the substrate solids after pretreatment did
not increase buffering capacity of the digestion(Ahn et al., 2009;
Kim et al., 2002).
The nal VFA concentrations for each experimental condition
tested are shown in Table 3. Acetic acid was the dominant component of VFA. For all the functional digesters, the VFA concentration
fell between 2.0 and 2.4 g/kg and the nal pH values were between
8.3 and 8.5, close to the initial values. In contrast, an accumulation
of VFA (22.1 g/kg) was observed for 7.5% NaOH-pretreated corn
stover at the end of the 20-day digestion, resulting in irreversible
acidication of digestion with a pH decreasing from 9.5 to 6.0.
The acetoclastic methanogens were reported to be severely inhibited by the accumulation of VFA and a decrease in pH (Powell and
Archer, 1986). Separated acidogenic and methanogenic phases
with leachate recirculation may provide buffering capacity and improve digestion performance (Myint and Nirmalakhandan, 2009;
Veeken and Hamelers, 1999).

60
50
40

untreated
1.0% NaOH

30

2.5%NaOH

20

5.0% NaOH
10
0
0

12

16

20

24

28

32

36

40

Time (d)
Fig. 3. Biogas production of simultaneous alkaline treatment and anaerobic
digestion: (a) daily biogas yield, (b) cumulative biogas yield, and (c) methane
content of biogas.

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J. Zhu et al. / Bioresource Technology 101 (2010) 75237528

to the anaerobic microbial community, especially hydrolytic and


acidogenic bacteria. As shown in Fig. 3c, the methane content of
the biogas increased to 5060% on day 7 for all the tests including
control. Only slight changes in methane content were observed
after day 16.
4. Conclusions
Solid-state alkali pretreatment for 24 h can effectively depolymerize corn stover with signicant lignin degradation and limited
cellulose loss. Solid-state anaerobic digestion of 5.0% NaOH-pretreated corn stover produced 37% more biogas when compared
to that of the untreated. But a higher NaOH concentration (7.5%)
caused inhibition of methanogenesis due to rapid hydrolysis and
acidogenesis. The simultaneous alkaline treatment and digestion
showed no signicant improvement on biogas production.
Acknowledgements
This project was supported by Ohio Agricultural Research and
Development Center (OARDC) Seeds Program. The authors would
like to thank Mrs. Mary Wicks (Department of Food, Agricultural
and Biological Engineering, OSU) for reading through the manuscript and providing useful suggestions.
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