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Biological Syngas Methanation via Immobilized Methanogenic Archaea on

Biochar

Article  in  Energy Procedia · October 2016

DOI: 10.1016/j.egypro.2017.03.396

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ScienceDirect
Energy Procedia 105 (2017) 823 – 829

The 8th International Conference on Applied Energy – ICAE2016

Biological syngas methanation via immobilized methanogenic

archaea on biochar
Sebastian Schwedea,*, Florian Bruchmannb, Eva Thorina, Mandy Gerberc
a
Mälardalen University, Box 883, 72123 Västerås, Sweden
b
Ruhr-University, Universitaestr. 150, 44780 Bochum, Germany
c
Bochum University of Applied Sciences, Lennershofstr. 140, 44801 Bochum, Germany

Abstract

Syngas containing H2, CO, CO2 and CH4 produced by thermal processes such as gasification or pyrolysis is typically converted to
methane via thermochemical methanation. This process is characterized by a high heat demand utilizing a sensitive chemical
catalyst at increased pressure conditions. Alternatively, methanogenic archaea could be exploited as a natural catalyst in a
biological methanation process with a lower energy demand. However, the mass transfer between the gas phase and the microbial
cell is a major challenge for efficient conversion of the syngas components. Therefore, in this work methanogenic archaea from
anaerobic digestion residues were successfully immobilized on biochar particles obtained from green waste pyrolysis with two
distinct particle sizes (0.25-1 mm and 1-2 mm). After incubation of the inoculated particles with an artificial syngas mixture CH4
was formed within the first 24 hours, while H2, CO2 and CO simultaneously declined. However, the particle size had no influence
on the CH4 yield, content and conversion efficiency. According to the maximum theoretical conversion rate of H2 with CO2 and CO
to CH4 only about 50% of the syngas components were converted to methane. These results suggest that CO was rather utilized by
the methanogens involved for acetate/formate formation than for methanogenesis due to slight inhibition of the latter process by CO
present in the syngas. The impact of CO inhibition during biological syngas methanation needs to be further evaluated for a
continuous application of the process. However, a proof of concept for this process using inoculated biochar particles could be
shown within the study presented here.

©©2017
2016The
TheAuthors.
Authors.Published
Publishedby
byElsevier
ElsevierLtd.
Ltd.This is an open access article under the CC BY-NC-ND license
Selection and/or peer-review under responsibility of ICAE
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Peer-review under responsibility of the scientific committee of the 8th International Conference on Applied Energy.

Keywords: biogas; syngas; methanation; biochar; anaerobic digestion; pyrolysis

1. Introduction
The Energy sector is responsible for the release of over 80% of the greenhouse gases (GHG) in the European
Union [1]. The majority of those emissions originate from both electricity and heat production and transportation.
Sweden, as one example in Europe, takes a pioneer role in mitigating the release of greenhouse gases as it generates
almost half of its primary energy from renewable sources [2-3]. However, in 2014 the proportion of renewable fuels
for transportation was only 12% of the total road traffic (5% in the European Union in 2013) [4]. As a measure to
increase renewable fuel production and utilization, the production of biogas from organic waste materials (i.e. sewage
sludge and source separated household waste) via anaerobic digestion (AD) is a common scenario for waste treatment
and energy recovery in Sweden [5]. More than 50% of the annual biogas produced is upgraded to biomethane for
utilization as vehicle fuel in public transportation busses.

* Corresponding author. Tel.: +46 (0)21 10 7360

E-mail address: sebastian.schwede@rub.de.

1876-6102 © 2017 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Peer-review under responsibility of the scientific committee of the 8th International Conference on Applied Energy.
doi:10.1016/j.egypro.2017.03.396
824 Sebastian Schwede et al. / Energy Procedia 105 (2017) 823 – 829

During AD polymeric organic compounds are enzymatically hydrolysed to soluble substances (hydrolysis),
fermented to various organic acids and alcohols (acidogenesis) and further degraded to mainly acetic acid, hydrogen
and carbon dioxide (acetogenesis) by a syntrophic bacterial consortium [6]. These metabolites are eventually
converted to methane by methanogenic archaea. Besides acetoklastic and hydrogenotrophic methanogensis (reaction
(1) and (2)) methanogenic archaea have been shown to utilize carbon monoxide (reaction (3) as well as methanol
(reaction (4)):

CH3COO- + H+ → CH4 + CO2 (ΔG0’=-36 kJ mol CH4-1) (1)

0’ -1
CO2 + 4H2 → CH4 + 2H2O (ΔG =-130 kJ mol CH4 ) (2)
0’ -1
4CO + 2H2O → CH4 + 3CO2 (ΔG =-211 kJ mol CH4 ) (3)
0’ -1
4HCO2H → CH4 + 3CO2 + 2H2O (ΔG =-106 kJ mol CH4 ) (4)

Not all organic biomasses that are collected at local waste treatment facilities are suitable for AD among others due
to unbalanced nutrient composition, potential toxic substances or a high degree of lignification. Lignin in particular is
not degradable under anaerobic conditions and results in reduced accessibility of the substrate to enzymatic
degradation [7]. Currently, most ligneous biomass is incinerated for heat or combined heat and power generation or
composted without any further energetic utilization. Alternative thermal conversion processes such as pyrolysis or
gasification enable the possibility to convert ligneous biomasses to CH4 to further increase the proportion of
renewable fuel. Dependant on substrate composition and process temperature, three main fractions are produced
during thermal conversion: oil, char and syngas. The syngas mainly consists of H2, CO2 and CO as well as traces of
CH4 and aromatic compounds [8]. To exploit the syngas as biofuel the gas requires conversion via a process referred
to as methanation.
There are two distinct ways for the methanation of syngas to a methane-rich gas. The first is thermochemical
methanation, which requires high temperatures (>250°C) and pressure as well as a catalyst, which is sensitive to
poisoning by impurities [9]. The second way is biological methanation via methanogenic archaea, which are able to
directly convert CO and CO2 with H2 to CH4 following reaction (2) and (3). The advantages of this process are
presumed higher energy efficiency as lower temperatures (35-70°C) and pressure are required. In biological
methanation the respective reactions are catalysed by microorganisms, which in contrast to the chemical process
tolerate impurities and loading changes of the substrate gas to a certain degree [10]. In addition, the conversion rate of
the syngas is higher and might reach methane contents of up to 98%, if the composition of the syngas is favourable
(i.e. high H2 content) [11].
The disadvantage of biological upgrading lies within the slower reaction rate compared to the chemical conversion.
For efficient biological syngas methanation a high contact area and mass transfer between gas and microorganisms is
required. Accordingly, the utilization of liquid phase reactors is a challenge due to the low solubility of especially
hydrogen in the liquid phase [12]. Alternatively, trickle bed reactors, wherein the microorganisms are immobilized on
a solid phase, while gas and liquid run in a counter current stream can be employed. Carrier materials successfully
employed in the biocatalytic methanation of hydrogen and carbon dioxide include polyethylene rings and
polyvinylidene fluoride sachets [11][13].
This paper is a proof of concept for biological syngas methanation utilizing biochar produced during the pyrolysis
of municipal solid green waste as a renewable carrier material. In the first place, the suitability of the biochar to
immobilize methanogenic archaea is tested. Secondly, the conversion efficiencies of inoculated biochar particles for
both the conversion of CO2 with H2 and CO to CH4 are evaluated under batch conditions at mesophilic temperatures.

2. Material and Methods

Biochar for the experiment was produced by pyrolysis of municipal solid green waste (total solids (TS): 55.2% of
fresh matter (FM); volatile solids (VS): 67.6% of TS) collected at the local waste treatment facility in Västerås,
Sweden. The green waste was shredded and sieved at the facility in two different fractions (>20 mm and <20mm). The
smaller fraction (<20 mm) was further dried at 105°C for 24 hours and utilized for pyrolysis at about 575°C and
 Sebastian Schwede et al. / Energy Procedia 105 (2017) 823 – 829 825

atmospheric pressure for one hour in a 5 L rocket stove. The biochar obtained after pyrolysis had a TS content of
98.8% of FM and a VS content of 38.7% of TS. The biochar was subsequently sieved to gain fractions of different
particle sizes. Two fractions were chosen for the experiment, particles sized 0.25 mm to 1 mm (fine biochar (FBC))
and 1 mm to 2 mm (coarse biochar (CBC)).
The fractions were incubated in a mixed digestate obtained from two mesophilic biogas plants, which process
mainly source separated organic household waste and sewage sludge, respectively. For incubation, biochar particles
were filled in nylon bags with a mesh size of 75 µm. The bags were placed in the digestate for 10 days at 38°C and
manually mixed once per day. After the incubation FBC and CBC showed a TS content of 31.9% and 26.0% of FM
and a VS content of 35.6% and 43.3% of TS, respectively. The incubated biochar was removed from the nylon bags
and samples were filled in glass bottles with a total volume of 1100 mL. Three bottles were filled with 130 g of fine or
coarse inoculated biochar, respectively. Separate fractions of inoculated biochar in both sizes were autoclaved at
140°C for 15 minutes to serve as a blank. After filling, the bottles were sealed gastight with butyl caps and aluminium
screw tops. For flushing and as substrate an artificial syngas mixture, which consisted of 48.4% H2, 26.4% CO2,
23.3% CO and 1.9% CH4 was used. Flushing of the headspace was performed for 2 minutes and the pressure in the
bottles was adjusted at 150 kPa before incubation at 38°C for 10 days.
During incubation, every 24 h the samples were shaken, pressure was measured and 20 mL of gas were taken for
analysis. Gas composition (H2, CO, CO2, CH4) was analysed using a gas chromatograph with a TCD detector at 130°C
(Compact GC, Interscience, equipped with a Shincarbon ST Micropacked 2m column, Restek). The column oven was
tempered at 120°C and helium was used as carrier gas at a flow rate of 20 mL min-1. The gas volume resulting from
the pressure in the bottles was obtained assuming ideal gas conditions. All gas volumes were corrected to standard
conditions (273.15 K; 101.325 kPa).
The yield and productivity of CH4 was calculated in relation to the amount of inoculated biochar added to the
sample (mL CH4 per g of biochar for the yield and mL CH4 per mL biochar and day for productivity). The theoretical
CH4 and CO2 proportion was calculated according to the conversion reactions (reaction (2) and (3)) with 1 mol CH4
formed for every 4 moles of H2 and CO assuming a full conversion of all components. For every mol CH4 formed
from 4 moles of H2 1 mol CO2 is converted, whereas for every mol CH4 from 4 moles of CO 3 moles CO2 are
released.

3. Results

Figure 1 shows the development of the respective syngas compounds (H2, CO2, CO and CH4) during the
mesophilic batch experiment with the two different sized inoculated biochar samples (Fig. 1A: FBC; Fig. 1B: 1-2
CBC). Both samples showed similar behaviour with decreasing amounts of H2, CO and CO2 and increasing amounts
of CH4 during the first 10 days of the experiment. Both H2 and CO were completely removed after 9 days. However,
the maximum CH4 productivity was obtained during the first 24 hours with 0.54±0.17 and 0.72±0.28 mL mL-1⋅d-1 for
CBC and FBC, respectively. The pressure and accordingly the gas volume dropped in all bottles with incubated
biochar during the course of the experiment. In all batch bottles with autoclaved biochar (blank) the concentrations of
all gas components and pressure remained stable within the error of measurement (data not shown).
826 Sebastian Schwede et al. / Energy Procedia 105 (2017) 823 – 829

Fig. 1. Development of all syngas compounds (H2, CO2, CO and CH4) during the mesophilic batch experiment with two different sized inoculated
biochar samples (A: FBC; B: CBC). Vertical error bars represent the standard deviation between triplicates.

In Table 1 the CH4 yield and the CH4 content for the biological syngas methanation experiment at mesophilic
batch conditions for the two different sized samples (CBC and FBC) are presented. In addition, the conversion
efficiency of the produced CH4 in relation to the theoretical maximum conversion of CO and H2 to CH4 is shown. As
for the volumes of the different syngas compounds (Fig. 1) both samples had similar results with differences within
the standard deviation of the triplicates. On average the CH4 yield was around 0.95 mL CH4 per g of inoculated
biochar with a final CH4 content of 41 Vol.% after 11 days. The conversion efficiency of CO and H2 to CH4 for both
inoculated biochar samples was around 50% of the theoretical maximum.

Table 1. CH4 yield and content and conversion efficiency of CO and H2 to CH4 in two different sized inoculated biochar samples evaluated from
mesophilic batch experiments.

Sample Biochar type CH4 Yield Conversion efficiency CH4 content

-1
(mL g biochar ) (%) (Vol.%)

1 CBC 0.99 49.81 41.5

2 CBC 1.00 53.44 41.6
3 CBC 0.87 47.64 41.1
Average CBC 0.95±0.07 50.30±2.93 41.4±0.2
1 FBC 0.88 43.18 40.7
2 FBC 1.04 51.48 41.4
3 FBC 0.89 43.32 40.9
Average FBC 0.94±0.09 46.00±4.75 41.0±0.3
CBC = coarse biochar, FBC = fine biochar

4. Discussion
After incubation of the inoculated biochar samples with the artificial syngas mixture at mesophilic temperature
conditions conversion of H2, CO2 and/or CO was evidenced within the first 24 hours by the formation of CH4. Apart
from that, no CH4 production was observed in the samples with autoclaved biochar during the whole experimental
 Sebastian Schwede et al. / Energy Procedia 105 (2017) 823 – 829 827

period and accordingly in contrast to the active samples gas pressure in these bottles stayed constant. This shows that
the biochar is a suitable carrier for methanogenic archaea and that the biochar was successfully inoculated with the
respective organisms during the 10-day inoculation phase in anaerobic digestion residues.
Furthermore, it could be shown that the size of the biochar particles did not influence parameters such as CH4 yield
and content or conversion efficiency during the biological methanation of the syngas. A higher conversion rate was
expected from the fine char, due to the presumed higher growth surface area for methanogens. Figure 2 shows a flow
sheet for the (theoretical) complete conversion of the syngas mixture in CBC and FBC for hydrogenotrophic (CO2
with H2, compare reaction (2) in introduction) and carboxydotrophic (CO, compare reaction (3)) methanogenesis and
the resulting CH4 and CO2 concentrations. A conversion efficiency of 100% for both pathways resulted in 254±6 mL
and 266±3 mL CH4 for CBC and FBC with a CH4 content of 36.1 Vol.%. In this case, about 2/3 of the CH4 produced
originates from the hydrogenotrophic pathway. However, only 50% (CBC) and 46% (FBC) conversion efficiency was
observed after 9 days, though 51% and 39% of CO and H2 were consumed on average already within the first 24 h and
both 100% after 9 days in CBC and FBC samples, respectively.

Fig. 2. Flow sheet for the theoretical complete conversion of the artificial syngas mixture to CH4 and CO2 with the gas amounts supplied in the
experimental batch test via hydrogenotrophic (CO2 with H2) and carboxydotrophic (CO) methanogenesis in CBC (A) and FBC (B).

An explanation for the low CH4 conversion efficiency could be the formation of acetate or formate from CO as
energy conservation for the methanogenic archaea [14]. This hypothesis is supported by the low amount and content
of CO2 during the whole experimental period. According to reaction (3) (see Introduction), 4 mol of CO are converted
to 3 mol of CO2 and 1 mol of CH4. However, the CO2 content decreased during the experiment. According to the
theoretical maximum conversion the final CO2 volume during the batch test was two times lower than the estimated
value. A complete conversion of all compounds to CH4 and CO2 results in a two times higher gas volume as observed
in the batch study performed.
Additionally, the expected CH4 and CO2 concentration in the gas is 36.1 Vol.% and 63.9 Vol.%, respectively.
Rother and Metcalf (2004) showed that acetate and formate were the dominant metabolic end products by
Methanosarcina acetivorans during growth on CO. Moreover, methane production declined with increasing CO
partial pressure due to inhibition of methanogenisis by CO. However, methane production exceeded acetate/formate
formation during the stationary growth phase [14] suggesting that after the reduction of CO both metabolites might be
utilized and converted to CH4. Nevertheless, this step makes it necessary to prolong retention time in technical
reactors, if CO is present in the syngas mix supplied.
In case the conversion of H2 (without CO conversion) is investigated as the sole source of CH4 generation, CH4
conversion efficiency increases on average to 74.5% (CBC) and 68.1% (FBC, data not shown). Moreover, the final
CH4 and CO2 volumes determined in the experimental batch test (Fig.1) were similar to the values obtained for the
theoretical complete conversion via the hydrogenotrophic pathway (Fig.2). This suggests that the main pathway for
828 Sebastian Schwede et al. / Energy Procedia 105 (2017) 823 – 829

methane production was hydrogenotrophic methanogenesis (reaction (2), see Introduction) and that CO was only
converted to metabolites others than CH4.
The relative CO concentration in the gas mix could be lowered by introducing extra H2 to the syngas. This would
further enhance CH4 production rate as H2 is the limiting factor in the hydogenotrophic pathway. Due to the low cost
and the high proportion of renewable electricity in Sweden, power-to-gas could be an option to produce H2 and
subsequently transform it to fuel during syngas upgrading. A second solution could be a biological pre-treatment of
the syngas, employing biologically catalysed water-gas shift reaction to form H2 and CO2 from CO and H2O [15].
Productivity could be further improved by enhancing char to gas contact area, e.g. in a trickle bed reactor.
Nevertheless, parameters such as CH4 yield and content and conversion efficiency as well as the energy efficiency in
comparison to the conventional chemical methanation process have to be further evaluated in long term studies with
continuous supply of substrate.

5. Conclusion
Biological syngas methanation using inoculated biochar particles was evaluated in a batch experiment at mesophilic
conditions. The results showed that the biochar was successfully inoculated with organisms showing methanogenic
activity. The conversion of H2 with CO2 and CO to CH4 was principally fast with more than 50% of conversion during
the first 24 hours. However, as shown by a reduced conversion efficiency CO was presumably utilized for
formate/acetate formation rather than for methanogenesis. The impact of this effect on a continuous methanation
process needs to be further evaluated.

Acknowledgements

The project has been performed as a co-production study within the framework Future Energy. The Knowledge
Foundation in Sweden (KKS), Vafab Miljö AB and Eskilstuna Energi och Miljö are thanked for their funding. For
their support with gas analysis the authors would like to thank the staff of Axel Semrau GmbH, Germany. The authors
would also like to thank the Foundation of Gustav Dahl (Stiftelsen Tandläkare Gustav Dahls Minne) for partly
supporting the work with a travel grant.

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Biography
Sebastian Schwede studied biology with focus on biochemistry, biotechnology and
microbiology at Ruhr-University in Bochum, Germany and obtained his diploma in 2009.
From 2009 he worked as a PhD candidate on the “Investigation and optimization of the
biogas potential of the marine microalga Nannochloropsis salina” at Ruhr-University
Bochum and obtained the PhD degree in mechanical engineering (Dr. Ing.) in 2013. The
doctoral studies were partly supported by a scholarship from the Graduate School of
Energy Efficient Production and Logistics and the Research School at the Ruhr-University. Since 2014
Sebastian Schwede works as a postdoctoral researcher at the Mälardalen University in Västerås, Sweden.
His main research areas include the implementation and optimization of biotechnological processes such
as biological waste water treatment, anaerobic digestion, biorefineries and biological methanation.

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