Renewable and Sustainable Energy Reviews 47 (2015) 23–33

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Renewable and Sustainable Energy Reviews

journal homepage: www.elsevier.com/locate/rser

Biocathode in microbial electrolysis cell; present status and

future prospects
Tahereh Jafary a, Wan Ramli Wan Daud a,b, Mostafa Ghasemi a,n, Byung Hong Kim a,
Jamaliah Md Jahim a,b, Manal Ismail a,b, Swee Su Lim a
a
Fuel Cell Institute, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia
b
Department of Chemical and Process Engineering, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

art ic l e i nf o a b s t r a c t

Article history: The application of the biocathode for hydrogen production in a Microbial Electrolysis Cell (MEC) is a
Received 26 August 2014 promising alternative to precious metal catalysts. However, biocathodes are still in the improvement and
Received in revised form development stages and require a deep understanding of the bioelectrochemical mechanisms involved.
8 January 2015
In this review, the results of biocathode MEC experiments and studies in the literature on biocathode
Accepted 1 March 2015
development methods were summarised; furthermore, used carbon sources and substrates in biocatho-
dic compartments and microbial communities on the biocathode were characterised. Based on the
Keywords: respective articles that were examined, biocathode MEC may be developed and initiated in one of three
MEC categories: (I) half biological two-chambered biocathode MEC; (II) full biological two-chambered
Biocathode
biocathode MEC; (III) full biological single-chambered biocathode MEC. In addition, various mixed
Hydrogen production
cultures capable of producing hydrogen were identified, and predominant species were detected.
Desulfovibrio paquesii, Desulfovibrio G11 and Geobacter sulfurreducens were also successfully tested as
pure cultures in biocathode MECs. Further studies are necessary for an acute and experimental
comprehension of the transfer of electrons and the energy conservation mechanism involved in the
biocathode MEC, which may provide a cost-effective and practical implementation of this technology.
& 2015 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
1.1. MFC to MEC development. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
1.2. MEC and biocathode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
2. Biocathode MEC configuration and start-up process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
2.1. Half biological two-chambered biocathode MEC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
2.2. Full biological two-chambered biocathode MEC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
2.3. Half biological two-chambered biocathode MEC and full biological single-chambered biocathode MEC comparative research . . . . . . . . 26
2.4. Challenges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
3. Carbon sources in biocathode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.1. Studied substrates regarding hydrogen production application in biocathode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
3.2. Studied substrate for wastewater application in biocathode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

n
Corresponding author. Tel.: þ 60 389118588; fax: þ 60 389118530.
E-mail addresses: mostafag@eng.ukm.my, mostafghasemi@gmail.com (M. Ghasemi).

http://dx.doi.org/10.1016/j.rser.2015.03.003
1364-0321/& 2015 Elsevier Ltd. All rights reserved.
24 T. Jafary et al. / Renewable and Sustainable Energy Reviews 47 (2015) 23–33

4. Biocathode microorganisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
4.1. Mixed cultures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
4.2. Pure cultures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
5. Future work . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
6. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

1. Introduction
Growing global energy demands are an inevitable issue due to the
high rate of population increase and fast industrial development.
Currently, the major portion of energy demand is provided with
conventional fossil fuel sources. Depletion and environmental pollu-
tion are two main problems associated with non-renewable fossil fuel
sources; therefore, current research is more motivated towards
locating renewable and clean sources of energy [1,2]. In this regard,
bioelectrochemical systems (BESs) have significant potential for
generating power, along with simultaneous wastewater treatment
[3–5]. Living cultures, as the environmental friendly biocatalysts,
displayed a sustainable role in bioenergy application [6]. Among the
discovered BESs, Microbial Fuel Cells (MFCs) and microbial electrolysis
cells (MECs) have attracted increased attention due to their ability to
produce bioelectricity and biohydrogen, respectively [7].
1.1. MFC to MEC development
It was in the late 18th century that the first empirical evidence of
energy production in the form of bioelectricity was approved by Luigi
Galvani via connection of frog legs to a metallic conductor. However,
the first MFC was fabricated by Potter in 1911 upon the illustration of
spontaneous current flow between an anode and a cathode electrode
applied in a bacterial culture (bioanode) and sterile medium (abiotic
cathode), respectively [8]. Promising sustainable energy production
along with wastewater treatment attracted increased scientific atten-
tion towards this technological development, significantly during
recent decades [9]. It was discovered that the current generation
reported by Potter was due to decreased redox potential during
bacterial growth. Kim et al. innovated the present MFCs which
employed electrochemically active bacteria, and were able to use the
electrode as the electron acceptor/sink in 1999; achieving the first
enrichment of microbial community to generate electricity in the MFC
basis electrochemical cell in 2004 [10,11].
Bioelectrochemical degradation of substrate (e.g. acetate) by
living microorganisms released protons and carbon dioxide into
anolyte and electrons to the anode as an oxidation half reaction in
a typical two-chambered MFC (Eq. (1)) [12]
C2 H3 O2  þ 2H2 O-2CO2 þ8e  þ7H þ ;
E1acetate=CO2 ¼  0:289 V vs:NHE at pH ¼ 7
Electrons flow through an external circuit to the counter
(cathode) electrode to produce bioelectrical current. Protons
diffuse across the separating membrane to the cathode where
they combine with electrons to form water in the presence of a
final electron accepter (e.g. oxygen) as the reduction half reaction
and complete the circuit (Eq. (2)) [12,13]
O2 þ4H þ þ 4e  -2H2 O;
E1H þ =H2 O ¼ þ 0:818 V vs:NHE at pH ¼ 7
In the absence of oxygen, H þ ions may be supported with an
extra applied voltage to be reduced to H2 in a modified MFC with
similar oxidation half reaction (Eq. (3)) [14]
H þ þ 2e  -H2 ; EH þ =H2 ¼  0:412 Vvs:NHE ð3Þ
Fig. 1 represented a schematic demonstration of MFC and MEC
systems with the components involved.
1.2. MEC and biocathode
The preliminary discovery of microbial hydrogen production in
a MFC basis reactor, named the Microbial electrolysis cell (MEC),
was reported by some researchers in 2005 (the so called technol-
ogy was microbial electrolysis cell) [15,16]. From an economic
perspective, it is believed that MECs are more environmentally
beneficial due to their functionality for producing chemical pro-
ducts (notably H2) when compared to MFCs with present insuffi-
cient power outputs [17].
Microbial electrolysis cells provide energy required for H þ to
H2 reduction (  0.412 V) via a microbial electrical supply
( 0.289 V), along with an external applied voltage. The extra
applied voltage is around 0.14 V theoretically, and more than 0.2 V
in practice when considering electrodes’ overpotentials and ohmic
losses; however, it is still significantly lower than what is needed
for water electrolysis (2.3 V) [18]. In addition, COD yields are
considerably higher in MECs (60–100%) compared to conventional
hydrogen production methods; e.g. steam reforming (63%), metha-
nol cracking (45%) and water electrolysis (19%). On the other hand,
hydrogen and carbon dioxide can be produced separately by
microbial electrolysis systems in contrast with phototrophic fer-
mentation which requires a two-stage process to produce a
mixture of these two gas products [19].
Expensive metal catalysts, such as platinum, are commonly
applied as the electrode catalyst to reduce overpotentials in MECs
and MFCs. Platinum had formerly displayed promising results in
conventional electrochemical systems due to its low activation
overpotential. In addition to cost, platinum is non-renewable,
scarce, subject to be poisoned by CO and sulphur and holds
negative effects on the environment [20]. The cathode plays an
important role in MEC systems where hydrogen is produced. The
ð1Þ cathode and its loaded metal catalyst contribute around 47% of the
total costs in MFCs [21]. The set up cost is a considerable, prevalent
and challenging factor (concern) for researchers in the field of
bioelectrochemical systems which will require a solution so as to
deliver these new technologies into a heightened level; out of the
labs [22]. Proposed drawbacks related to applying metal catalysts
prompted current studies to focus more on cost effective catalysts
[7,23–27]. Favourably, from among others, microorganisms have a
special rate of interest that made them an inexpensive alternative
since they are environmentally friendly, hold a self-generative
character and are resistant to certain levels of impurities such as
ð2Þ sulphur [28]. In a biocathode MEC, microorganisms are able to use
the surface of the electrode as an electron source to catalyse the
reaction of electrons and protons to form hydrogen.
 T. Jafary et al. / Renewable and Sustainable Energy Reviews 47 (2015) 23–33
e- e-
V
CO2
Out H H2O
CO2
H
Acetate H
Feed In
Anode Cathode
Membrane
e e
PS
CO
Out
H
CO
H
Acetate H H
Feed In
Anode Cathode
Membrane
Fig. 1. Schematic overviews of MFC and MEC by demonstration of standard potentials (vs. NHE) of oxidation and reduction reactions in anodic and cathodic chambers,
resulting in a spontaneous reaction in MFC and a nonspontaneous reaction in MEC.
Improving the functionality of the biocathode as a promising
alternative to precious metal catalysts in terms of overpotential,
hydrogen production yield, start-up time and cathodic hydrogen
recovery may lead this new encouraging technology into an
industrial scale by focusing on hydrogen production from a wide
range of renewable sources with low capital cost. This paper
reviewed the biocathode development procedures, biocathodic
nutrient sources, as well as, microbial communities that were
applied by different scientists in published biocathode MEC
research articles thus far.
2. Biocathode MEC configuration and start-up process
Similar to the configuration of MFCs, MECs consist of an anode,
a cathode and an optional separating membrane [29]. Employing
the membrane is preferred due to the advantage of producing CO2
and H2 gases separately in the anodic and cathodic chambers,
respectively. In addition, there is no propitious condition for
methanogens to consume H2 gases in the presence of the mem-
brane in an abiotic cathode MEC. Additionally, a power source is
essential to supply energy required for non-spontaneous reactions
(hydrogen production) in MECs in contrast to spontaneous reac-
tions (water formation) in MFCs (Fig. 1). A few start-up procedures
were applied by various researchers to initiate a biocathode MEC,
the most common of which were mentioned in this paper. The
first procedure involved a full biological start-up with electro-
chemically active microorganisms in both compartments of a two-
chambered MEC and/or in a membraneless one-chambered MEC.
The second procedure consisted of a half (cell) biological start-up
by applying the electrode polarity reversal to change a bioanode to
the biocathode and/or transferring the active culture, or bioelec-
trode, of another MEC to the cathode chamber without polarity
reversal; both with an abiotic anode. In other words, there are
three categories of biocathode MEC based on the start-up and
development process reported in the biocathode MEC literature so
far: (I) half biological two-chambered biocathode MEC; (II) full
biological two-chambered biocathode MEC; (III) full biological
single-chambered biocathode MEC (Fig. 2).
25
E vs. NHE(mV)
O2
+818
O2 /H 2 O
H2O
O O Spontaneous v oltage
generating reaction in MFC
0 0
H
-289
Acetate/CO 2 Nonspontaneous power -source
H required reaction in MEC
- 414
H+/H 2
Anode Cathode
Fig. 2. Different start-up methods of MEC biocathode using one or two-chamber
reactors. Category I: half biological two-chambered biocathode MEC. Category
II: full biological two-chambered biocathode MEC. Category III: full biological
single-chambered biocathode MEC.
2.1. Half biological two-chambered biocathode MEC
The first microbial-origin biocathode MEC was established through
a three step start-up procedure in a two-chambered bioreactor
(Category I, Fig. 3i): (a) enriching an acetate-fed bioanode with
electrochemically active mixed culture, accompanied by flushing the
headspace with H2; (b) replacing the acetate with sodium bicarbonate
and persistent hydrogen flushing; and (c) reversing the polarity of the
electrodes accordingly (Fig. 3ii). Microorganisms were allowed to grow
in the first 50-h batch mode to prevent the wash-out phenomena
before switching to the continuous flow of the nutrient medium.
Around 250 h after inoculation, the biocathode MEC was achieved by
performing a polarity reversal scan in advance. The biocathode reached
to 1.1 A/m2 of current density at  0.7 V potential, which was
2.4 times the current density obtained in another study by titanium
electrode coated with platinum [30]. Promising results were shown by
comparing the potential applied for the same value of the current
density ( 0.47 A/m2) for platinum coated (with  0.7 V) and bio-
cathode (with  0.65 V) MECs. Hydrogen yield tests demonstrated
0.63 m3 H2/m3/day which was subjected to 67–94% of hydrogen loss,
mostly due to hydrogen diffusion through the membrane. Carbon
limiting was applied to halt hydrogen consumption by methanogens
upon removing bicarbonate from the nutrients before hydrogen tests.
Pisciotta et al. [31] applied a novel method to develop a biocathode
originating from the bioanode of a sediment microbial fuel cell without
the need for set potentials or chemicals, such as ferricyanide, used in
earlier bioanode to biocathode inversed study. The choice of sediment
MFC was to provide an anaerobic condition comparable to typical
26 T. Jafary et al. / Renewable and Sustainable Energy Reviews 47 (2015) 23–33
Fig. 3. (i) Schematic of the first biocathode MEC experimental set up. “Reprinted with permission, Copyright (2014) American Chemical Society”; (ii) Three-step biocathode
development procedure in the first half biological biocathode MEC: (a) enriching an acetate-fed bioanode with electrochemically active mixed culture accompanied by
flushing the headspace with H2, (b) replacing the acetate with sodium bicarbonate and persistent hydrogen flushing, (c) reversing the polarity of the electrodes accordingly.
Ferocyanide/ferricyanide was used as an electron donor and acceptor. “Reprinted with permission, Copyright (2014) American Chemical Society” [25].
electrode suspension in the electrolyte that was more susceptible to
dissolved oxygen contamination through gaskets and seals, or media
replacements [31]. Table 1 displayed the experimental conditions and
obtained results of various biocathode MECs reported in the literature
so far.
2.2. Full biological two-chambered biocathode MEC
Although the first half cell biological biocathode MEC research
approved the application of microorganisms as the cathode catalyst, it
still functioned under half biological conditions (bioanode and abiotic
cathode in the first two steps, and biocathode and abiotic anode after
polarity reversal in the third step). Jeremiasse et al. [41] studied the first
full biological electrolysis cell in which both oxidation and reduction
reactions were biocatalysed with electrochemically active microorgan-
isms (bio-cathode and anode) in the same experimental setup as in
[25] (Category II). The biocathode was inoculated 600 h after the
establishment of the bioanode from previous polarity reversed experi-
mental setups [25]. Current density of 3.3 A/m2 was obtained at the
cathode potential of  0.7 V (applied voltage of 0.8 V) which was
comparable with that of a continuous membraneless MEC of 3.2 A/m2
catalysed with 5 g/m2 load of platinum [42]. However, the result was
far from the current density of 11.7 A/m2 in another membraneless
MEC operated in the batch mode and coated with 5 g/m2 load of
platinum [43]. Low hydrogen rate of 0.04 N m3/m3/day and cathodic
hydrogen recovery of 21% were reported despite applying a carbon
limiting condition, which were lower than those obtained in the same
experimental setup for the first half biological biocathode MEC
(0.63 m3 H2/Volume/day and 49%, respectively) [25] (Table 1).
2.3. Half biological two-chambered biocathode MEC and full
biological single-chambered biocathode MEC comparative research
As a comparable study, biocathode MEC was investigated in an
inverted bioanode half cell (Category I) after substrate depletion
and was compared to an identical membraneless MEC (Category
III) in the same study that operated with applied voltage from the
start (Fig. 4i). The highest hydrogen production rate of about
24 mmol/h was reported at a cathode potential of 1.0 V with 56%
cathodic hydrogen recovery in the biocathode MEC, and with
ferricyanide in the anode. The results for membraneless MEC at a
similar cathode potential of 1.0 V (0.7 V applied voltage) were
10.8 mmol/h of hydrogen production rate and 36% of cathodic
hydrogen recovery. However, it was unclear whether any biocata-
lytic (biocathode) activity by microorganisms in one compartment
of MEC was present in this research [15]. Fu et al. [37] investigated
the biocatalytic activity of the cathode by developing one and two-
chambered biocathode MEC without polarity reversal (Categories
III and I, respectively; Fig. 4ii). It was the first alternative method
for developing a biocathode without reversing the polarity of the
electrodes. First, the biocathode was developed in a membraneless
MEC by inoculating from a three months advanced in operation
MFC, and then investigated for biocatalytic activity of the electro-
des (Category III). Afterwards, the cathode of the membraneless
setup was placed as the biocathode of the two-chambered MEC
(“C-C”). In addition, the anode of the membraneless setup was also
placed as the biocathode (“A-C”) to compare results with those
from polarity inverted bioanodes to biocathodes (category I). The
abiotic anode with oxidation of potassium ferrocyanide was
applied. Cyclic voltammetry proved catalytic ability of the anode
for acetate oxidation, and higher catalytic activity of the cathode to
produce H2 in comparison to the anode in the single-chambered
experiments. By performing further analysis in the two-
chambered setup, large cathodic currents were obtained at the
potential of 0.65 V in C-C design compared to small amounts
recorded in the A-C design at the potential of less than 0.7 V
[37]. Similar to previous comparable studies [15], lower cathodic
H2 recovery was obtained for the membraneless setup (20%)
compared to the two-chambered biocathode MEC (70%). At
cathode potential of  0.8 V, current density of  1.28 A/m2 and
hydrogen production rate of 376.5 mmol/day/m2 were recorded
for the C-C configuration (Table 1).
2.4. Challenges
Hydrogen lost across the membrane was a considerable fact that
maintained both membraneless and two-chambered configuration
and were attractive research subjects. Although using a membrane in
an abiotic cathode MEC was preferred due to the prevention of
methanogens to consume H2 in the product chamber, the membrane
had other aspects that required study. Although, there was the
potential of methanogenisis in biocathode MEC, nonetheless. Further-
more, applying the membrane presented high hydrogen loss and
scaling problems; however, membraneless setups resulted in low
cathodic H2 recovery which was probably due to the utilisation of H2
products by exoelectrogens to produce electricity on the anode. On
the other hand, full and half biological biocathode had their own
issues that were not yet addressed or completely studied. Scaling and
bioanodic associated limitations were prevented in the full biological
two-chambered biocathode setup, while the two-chambered was
able to produce higher current density. A profound understanding of
 T. Jafary et al. / Renewable and Sustainable Energy Reviews 47 (2015) 23–33 27

Table 1
Overview of experimental conditions and obtained results for various biocathode MECs reported in the literature.

Biocathode Biocathode type Biocathode catalyst Membrane Cathodic Mode of Cathode Hydrogen Hydrogen Current Ref.
start-up chamber operation potential production recovery (A/m2)
process size (ml) (V) rate (m3 H2/ (%)
m3 reactor/
day)

Category Ia Graphite felt, Active mixed culture taken from an CSMd 250 Continuous,  0.7 0.63 49  1.2 [25]
S.Ab ¼250 cm2, active bioelectrochemical cell 1.3 ml/min
Thc ¼ 6.5 mm
Non-porous flat Pure culture of G. sulfureducens CEMe 33 Continuous, 1 0.43 56  2.4 [32]
graphite in contact filtered and re-suspended in 0.85 l/h (24 mmol/h)
with graphite felt biocathodic cell
filled in chamber
S.A¼ 22 cm2
Graphite felt, Active culture taken from a four-year CSM 250 Continuous  0.7 0.63 – 1.2 [33]
Th¼ 6 m culture enriched in MFC and MEC
Graphite felt, Pure Culture of Desulfovibrio G11 CEM 250 Continuous  0.7 – – 0.76 [33
Th¼ 2.5 m 60 ml/min
Graphite rod Pure culture of Desulfovibrio paquessi PEMf 270 Batch  0.9 0.12 3 [34]
S.A¼ 9.7 cm2
D¼ 6 mm
Brush Graphite Transferred biocathode from a CEM 190 Batch  0.539 0.08 mmol –  0.45 mA [31]
fibre S.A ¼0.22 m2 sediment MFC
Graphite granules Brewery wastewater PEM 150 Batch  590 0.295 – – [35]
30 g (dry weight) (11.8 mM/
day)
Graphite granules From previous electrosynthetic cell PEM 150 Semi batch  590 2.5 2.5 – [36]
30 g (dry weight) that originally was inoculated with (100 mM/
brewery wastewater day)
Plain carbon cloth Transferred Biocathode from PEM 300 Batch  0.8 376.5 mmol/ 70  1.28 [37]
S.A¼ 40 cm2 previous single chambered day/m2
biocathode setup inoculated with
mesophilic microorganisms.
Graphite paper, Effluent of a previous operated MEC CEM 33 Continuous  0.7 – – 1 [38]
S.A¼ 22 cm2 36 ml/h
Graphite felt, Effluent from a MEC biocathode CEM 100 Continuous  0.7 2.4 – 2.7 [38]
S.A¼ 100 cm2, 156 ml/h
Th¼ 0.25 cm
Filled with A mixed effluent of urban CEM 390 Continuous  0.9 0.1 – 17A/m3 [39]
granular graphite wastewater treatment plant and HRT¼ 6.25
along with from a MFC used for treating waste
graphite electrode
S.A¼ 0.052 cm2
Graphite felt Effluent from a 30-day in run CEM – Continuous  0.7 2.2 50 2.7 [28]
S.A¼ 100 cm2 biocathodes, which were inoculated 60 ml/min
with a culture collected from UASB
reactor and enriched in anodes of
MECs over 5 years

Category IIa Carbon paper Dechlorinating cultures maintained PEM 270 Batch  0.75 0.01 – 4.4 [40]
S.A¼ 8 cm2 in anaerobic reactors
Graphite felt, Effluent from the biocathode in Ref. CSM 250 Continuous,  0.7 0.04 21 3.3 [41]
S.A¼ 250 cm2, [25] 1.3 ml/min
Th¼ 6.5 mm

Category IIIa Plain carbon cloth Effluent from a thermophilic three- – 250 batch  0.8 0.46 mmol/ 20 -0.87 [37]
S.A¼ 40 cm2 month in operated MFC day/m2
Graphite electrode Pure culture of G. sulfureducens  33 Continuous  1.13 0.31 43 1.21 [32]
filtered and re-suspended in (17.2 mmol/
biocathodic cell h)

a
Category I: half biological two-chambered biocathode MEC; Category II: full biological two-chambered biocathode MEC; Category III: full biological single-chambered
biocathode MEC.
b
S.A ¼surface area.
c
Th¼ Thickness.
d
Cation selective membrane.
e
Cation exchange membrane.
f
Proton exchange membrane.

the microorganisms involved in H2 formation and electron uptake
mechanism in the biocathode may help to enhance the biocathode
MEC in both one and two-chambered design; based on the applica-
tion and condition. Table 2 summarised relevant issues relating to
three mentioned categories discussed in published articles regarding
biocathode MECs.
3. Carbon sources in biocathode
Although bioelectrochemical systems (such as MECs) recorded
lower product yields compared to chemical methods, they are still
considered to be a promising technology due to their potential to
use renewable resources and wastewater as a wide range of
28 T. Jafary et al. / Renewable and Sustainable Energy Reviews 47 (2015) 23–33

Fig. 4. (i) Schematic of two-chambered MEC converted to a biocathode MEC for testing the ability of Geobacter sulfurreducens for hydrogen production in biocathode MECs.
“Reprinted with permission. Copyright (2014) American Chemical Society” [32]; (ii) (A) single and (B) two-chambered MEC setups in which bioanode and biocathode of
single chambered MEC were transferred to two-chambered as the biocathode in two separate experiments. “Reprinted with permission. Copyright (2014) International
Journal of Hydrogen Energy (IJHE)” [37].

Table 2
The pros and cons reported for three categories of biocathode MEC, along with predicted discussed solutions and reasons.

Category Advantages Reported problems Cathodic and anodic reaction Reasons Suggested solutions
after biocathode development

Category 1. Low scaling 1. Consumption of Anodic: ([Fe(CN)6]4  -[Fe(CN) 1. Hydrogen loss by diffusing through 1. Removing the carbon
I effects hydrogen by 6]3  þ e  ) Cathodic: the membrane source from cathodic
2. No possible hydrogenotrophic (CO2 þ H2O2H2CO32H þ þ HCO3 ) medium (carbon limiting
limitation by methanogens [25] [37] policy) [25]
a bioanode 2. Low cathodic 2. Using a thicker membrane,
[28] hydrogen or other membrane
recovery [25] materials
3. Providing higher current
density that produce more
hydrogen, while diffusional
hydrogen loss are constant
[25]

Category 1. High current 1. Low hydrogen – 1. Hydrogen loss is a result of H2 1. Higher current density,
II density [41] recovery [41] diffusing through the membrane higher hydrogen recovery
2. Methane and tubing 2. More extensive research on
detection in 2. Methane may have been formed by the type of the membrane
biocathode methanogens present in the is required
chamber despite bioanode, and subsequently crossed
carbon via the membrane to the biocathode
limiting policy 3. Methane could have been formed by
3. Current density methanogens present in the
drop after long biocathode from hydrogen and
time of operation carbon dioxide (originating from the
[41] bioanode)
4. Hydrogen loss 4. Precipitation of calcium phosphate
on the biocathode

Category 1. No scaling, 2. Low cathodic H2 – 1. H2 produced in the single- -

III resistance recovery [37] chambered MEC was oxidised by the
and loss due 3. Contamination of exoelectrogens on the anode which
to absence of cathodic product led to electricity generation [37]
membrane with anodic gas
product

feedstock. In addition, BESs are cost effective and can reduce
energy demands. From this aspect, biocathode MEC are appreci-
able since they are less susceptible to poisonous components
present in real wastewaters or natural resources, in comparison
to metal catalysts. To develop a biocathode MEC, a carbon source is
necessary to supply cathodic biocatalyst growth; furthermore,
they have shown effective impact on main/side product formation.
The substrates used in different biocathodic electrolysis systems
were reviewed by considering two discussed applications: hydro-
gen production and wastewater treatment.
3.1. Studied substrates regarding hydrogen production application in
biocathode
In the first biocathode MEC, sodium acetate was replaced by
sodium bicarbonate before bioanode conversion to the biocathode.
No hydrogen was detected and methane was the only product. By
assuming that the bicarbonate served as a carbon source for
hydrogenotrophic methanogens for consuming hydrogen, carbon
limiting strategy was applied by removing the bicarbonate before
hydrogen tests. Authors claimed that they preserved this strategy for
 T. Jafary et al. / Renewable and Sustainable Energy Reviews 47 (2015) 23–33
more than 1000 h without effecting the current generation in the
biocathode system [25]. However, the carbon limiting strategy was
not efficient in the full biological two-chambered MEC, fed with
sodium bicarbonate in the cathode medium and sodium acetate in
the anode medium. It was reported that detected methane may have
been produced in the bioanode and then diffused across the
membrane to the cathode; or methanogens in the biocathode could
have utilised CO2 of the anode and H2 of the cathode chambers to
form methane [25,41]. In addition, a biocathode requires a long
adaption and start-up time, along with low production rate, in
comparison to metal catalysts. It was reported that the replacement
of bicarbonate as an autotrophic, with acetate as a heterotrophic-
carbon source, has improved the start-up procedure’s time duration
to double; and further improved the H2 production rate to seven
times, about 2.2 m3 H2/m3 reactor/day compared to previous studies
[25,28]. This was the highest level of hydrogen production rate in
the biocathode MEC so far. Additionally, the sulphate element was
removed from the media to stop growth of sulphate reducing
microorganisms in this research. No methane and hydrogen sul-
phide were detected. In a recent study, Jeremiasse et al. [38] had
experimented with biocathode MECs to determine how carbon
sources will affect the growth and microbial communities in the
biocathode development process. Acetate, bicarbonate and acetate
without sulphate-fed MECs were studied in five experimental
setups. Although applying acetate as the carbon source displayed
half the start-up time compared to bicarbonate (28 days to 63 days,
respectively), insignificant results were obtained regarding the
average current and H2 yield; irrespective of carbon source. Further-
more, it was concluded that the design had a considerable influence
on the biocathode development and its biofilm composition [38].
In another biocathode study performed by Marshal et al.,
carbon dioxide presented a capability as the sole carbon to
produce hydrogen at the rate of 11.8 mM/day at cathode potential
of  590 mV; aside from acetate and methane as two other co-
products in an electrosynthetic system [35]. The results improved
to around 100 mM/day at the same cathode potential of  590 mV
in a similar experimental setup operated in a semi batch mode
over 150 days in a later study [36]. Table 3 summarised the
biocathode electrolysis experiments by considering the used
carbon sources, and analysed microbial communities that may
have been affected by carbon sources as one of the effective
parameter.
3.2. Studied substrate for wastewater application in biocathode
Wang et al. [44] investigated the toxic nitrobenzene waste-
water (NB) conversion to a less poisonous product of aniline in a
biocathode BES in the presence of glucose and bicarbonate at 0.5 V
of applied voltage. While nitrobenzene reduction presented a 10%
decrease by replacing glucose to bicarbonate, selective reduction
of nitrobenzene to aniline at a high rate of 98.93% was never-
theless achieved. Nitrobenzene reduction in this biocathode sys-
tem was considerable compared to earlier NB reduction studies in
an abiotic cathode and MFC system with pt loaded cathode; in
regards to the overpotential, precious metal catalyst application
and chemical costs [8,45,46]. Hydrogen measurement was not the
focus of this research; however, hydrogen was studied to see
whether it can play an electron donating role for nitrobenzene
reduction in the system. The research results suggested that the
autotrophic condition may not be necessary and developing the
biocathode was possible with glucose as an organic carbon source
that is generally available in the real environment [44]. During a
subsequent research in 2014, Liang et al. [47] increasingly studied
the NB reduction to aniline by switching organic carbon source
(glucose) to an inorganic source (bicarbonate) in a biocathode BES.
They focused on: (a) the effect of carbon source exchange on
29
biocathode catalytic properties and (b) further investigation of
microbial communities in biocathode biofilms as a result of carbon
source switchover. Eight reactors were employed to handle
experiments in fed batch modes; five were running under glucose
with different concentrations, two were focusing on carbon source
switchover experiments (glucose to bicarbonate), while the other
was kept operating with the abiotic cathode for trustworthy
comparison purposes. Four various modes were considered for
the NB reduction process: mode—(a) glucose-fed biocathode;
mode—(b) glucose-to-bicarbonate switchover to biocathode;
mode—(c) open biocathode; mode—(d) abiotic cathode. NB never-
theless continued reducing to aniline after carbon source switch-
over, however, with a lower NB reduction and aniline formation
rate [47]. The results were consistent with those reported in
previous studies [44]. Authors had performed detailed research
on the effect of carbon switchover regarding the biocathode
microbial community which will be discussed later in this paper.
Azo dye could also be successfully removed in a single-chambered
biocathode electrolysis system at 0.5 V of applied voltage. Biocathode
setup showed 81.7% of removal efficiency in just 10 h, which was
around 13% higher than what was achieved in an abiotic cathode [48].
In another research study, a type of antibiotic-content wastewater
was also treated in bio and abio-cathode electrolysis systems, and the
cathodic reduction process was investigated (at 0.5 V of applied
voltage). Higher current peaks, lower overpotential and less toxic
intermediates were observed at the biocathode in comparison to
those in the abiotic cathode. Glucose was used as a carbon source to
donate electron intracellularly, while the electrode fulfilled an extra-
cellular electron donation role (Table 3) [48].
4. Biocathode microorganisms
When applying living cultures as the biocatalysts on the cathode,
there are some basic requirements that microorganisms must satisfy in
order to catalyse half reduction reactions. Biocatalysts should be
capable of overcoming thermodynamic limitation to form hydrogen
by externally applied voltage. In addition, the electrode surface should
be utilised as an electron source (donor) by biocatalysts through the
biocatalytic activity of pure or varied species of mixed cultures to
reduce H þ ions to hydrogen, CH4 and other expected products [32,49].
Rosenbaum et al. extensively reviewed the possible Extracellular
Electron Transfer (EET) mechanisms through published articles on
the biocathode, as well as, known mechanisms for biological processes.
Finally, they suggested that c-type cytochrome, along with hydroge-
nases, would be engaged in bioelectrochemical processes for direct
electron transfer [50]. Hydrogenases are the enzymes involved in
bioelectrochemical systems to catalyse H2 production/consumption
[51–53]. There are three categories for hydrogenases based on their
metal sites that are active for a reduction reaction: nickel–iron, iron–
iron and iron–hydrogenases (Ni–F, Fe–Fe and Fe–hydrogenases, respec-
tively) [54,55]. However, using purified hydrogenases include the
related drawback of low stability and catalytic activity during long
durations, which can be improved by using whole cells [33]. MECs
suffer from a lack of knowledge regarding the biocatalytic hydrogen
formation mechanism, the type of involved hydrogenases in H2
evolution, active microorganism on the biocathode, electron uptake
mechanism from the bioelectrode and energy management by the cell.
4.1. Mixed cultures
The first mediatorless biocathode MEC was developed by using
natural mixed culture with microbial origin. The microbial origin-
ality of the microorganisms was investigated by the inhibition test
(by exposing the biocathode to carbon monoxide), along with
subsequent inoculation to a new biocathode setup; however,
30 T. Jafary et al. / Renewable and Sustainable Energy Reviews 47 (2015) 23–33

Table 3
Summary of the dominant microorganisms identified in biocathodic electrolysis cells.

Culture source Hydrogen Dominant species Carbon Biocathode Detected Analysis used for Ref.
revealed source development Products biofilm
cathode procedure characterisation/
potential study

Mixed culture Brewery  590 Acetobacterium spp, CO2 Biocathode origina Methane, CV, RNA [35]
wastewater Sphingobacterials, acetate extraction, RT-
Methanobacterium (Firmicutes, and PCR
Bacteroidetes, Proteobacteria, hydrogen amplification, 16S
Deferribacteres, Synergistetes and rRNA sequencing,
Spirochaetes at phylum level) SEM
Marine sediments o  400 Autotrophic: Eubacterium limosum, No added Bioanode of the Methane, LSV, DNA [31]
Desulfovibrio sp.A2, Rhodococcus, organic sediment MFC hydrogen extraction and
Gemmata obscuriglobus carbon after 16S rRNA gene
biocathode amplification,
start-up Gram staining
Dechlorinating  450 (with Desulfitobacterium spp., No organic Biocathode origin Hydrogen Eubacterial probe [40]
bacteria redox dehalococcoides spp. carbon
mediator), source
 700
(without
redox
mediator)
A mixed affluent of o  900 Hoeflea sp., Aquiflexum sp. Bicarbonate Biocathode origin Hydrogen DNA extraction [39]
urban wastewater and 16S rRNA
treatment plant and gene
from a MFC used for amplification,
treating waste SEM, DNA
extraction
Activated sludge  0.74 Enterococcus sp. (Firmicutes at Glucose & Biocathode origin Hydrogen, DNA extraction [44]
phylum level) bicarbonate aniline and 16S rRNA
gene
amplification
Microbial origin  0.7 Rhodopseudomonas palustris Bicarbonate Reversed bioanode Hydrogen DNA extraction [49]
mixed culture (Proteobacteria at phylum level) and 16S rRNA
gene
amplification
Activated sludge – Enterococcus (Firmicutes at phylum Glucose Biocathode origin Aniline Gene array, DNA [47]
level) extraction and
Paracoccus & Variovorax Bicarbonate 16S rRNA gene
(proteobacteria at phylum level) amplification, CV
Thermophilic  0.7 Firmicutes phylum No added Biocathode of a Hydrogen DNA extraction [37]
microorganism carbon single-chambered and 16S rRNA
from a previous source after setup placed as the gene
MFC setup biocathode biocathode of the amplification, CV
start-up dual-chambered
setup
Effluent of a  0.7 Clostridium cylindrosporum, Acetate, Biocathode origin Hydrogen SEM, DNA [38]
previous operated Desulfotomaculum sp., Clostridium bicarbonate extraction and
MEC cylindrosporum, Desulfotomaculum 16S rRNA gene
sp. Hydrogenophaga flava, Azonexus amplification,
caeni (At phylum level: Firmicutes DNA microarray
for small setups and proteobacteria analysis
and bacteroidetes for large setups,
regardless of carbon source)
Effluent of a  0.7 Desulfovibrio vulgaris (protebacteria Bicarbonate Bioanode inversed Hydrogen DNA extraction [33]
previous operated at phylum level), Desulfitobacterium biocathode and 16S rRNA
MEC and MFC hafniense (Firmicutes at phylum gene
level) Rikenella microfusus amplification,
(Bacteroidetes at phylum level) clone library
analysis, DGGEb

Pure culture Pure culture of G.  0.8 – No added Bioanode inversed Hydrogen – [32]
Sulfurreducens carbon biocathode
source after
biocathode
start-up
Pure culture of  900 – Bicarbonate Biocathode origin Hydrogen – [34]
Desulfovibrio
paquesii
Pure culture of  700 – Bicarbonate Biocathode origin Hydrogen – [33]
Desulfovibrio G11

a
Biocathode was developed by culturing the cathode directly with microorganisms in the cathodic compartment from the beginning.
b
Denaturing gradient gel electrophoresis.
 T. Jafary et al. / Renewable and Sustainable Energy Reviews 47 (2015) 23–33
microbial active species and mechanism were not analysed and
studied [25]. Following the proof-of-principal of a full biological
two-chambered MEC by Jeresmiasse et al. [41], microbial diversity
of the bioanode and biocathode was scrutinised in a MEC devel-
oped with similar start-up procedures as Rozedal et al. [25] and
Jeresmiasse et al. [41]; it was inoculated with a mixed culture [49].
Two clones associated with Bacteroidetes and Firmicutes phyla
were identified in the bioanode which were previously reported
in anodes of various MFCs [56,57]. Rhodopseudomonas palustris
was assessed as the dominant species in the cathodic bioelectrode
which had Ni–Fe and Fe–Fe–hydrogenases, and was a known
species in hydrogen production processes [57–60]. In an extensive
experimental research, Croese et al. [38] characterised the bacter-
ial diversities of five biocathode MECs that were different in size,
electrode material, flow path and carbon source. Firmicutes in
small MECs, and proteobacteria and Bacteroidetes in large MECs,
were classified as prevalent groups in both acetate and bicarbo-
nate fed MECs but with various percentages. Actinobacteria by 98%
was revealed as the dominant associated group in the large MEC
fed with acetate by excluding sulphate from the media. Regardless
of the carbon sources used in small setups, Clostridiaceae and
Peptococcaceae were identified as the dominant ribotypes. Bacter-
oidetes and Betaproteobacteria were the prevalent ribotypes for
acetate and bicarbonate large fed MECs, respectively. The differ-
ences between dominant species in large setups were more
elaborately investigated by hydrogenases chips in this research.
Ni–Fe and Fe–Fe hydrogenases and reducing hydrogenases coen-
zyme F420 were abundantly located in acetate fed biocathode
MEC. Various Ni–Fe hydrogenases were the ample genes in
bicarbonate-fed setups. Detailed information on dominant ribo-
types, established hydrogenase gens and various phyla distribu-
tions were reported for all 5 experimental setups in this research
article. Nevertheless, the effect of acetate or bicarbonate-fed MECs
in this research study remained unclear, while no exclusive
heterotrophic/autotrophic in respect to bacteria were distin-
guished [38]. However, glucose and bicarbonate feedstock resulted
in two different heterotrophic and autotrophic dominant species
in other biocathode electrolysis setups of nitrobenzene reduction
research. Enterococcus in glucose-fed and Paracoccus and Vario-
vorax in bicarbonate-fed biocathode systems were identified as the
dominant species [47]. Substantial performance achieved by
thermophilic microorganisms in some MFCs setups [61–64] pro-
vided the idea for the first MEC biocathode which was developed
with thermophilic microorganisms in 2013 [37]. Analysing ther-
mophilic biocathode microbial community revealed 21 phylotypes
which belonged to six phyla; dominated by the Firmicutes phylum.
Hydrogenophilic dechlorinating cultures (desulfitobacterium and
dehalococcoides-enriched cultures) may also catalyse H2 produc-
tion in biocathode MECs with methyl viologen at the cathode
potential of  450 mV, and without mediator at potentials less
than  700 mV [40].
4.2. Pure cultures
Other than mixed cultures, Geobacter sulfurreducens and Desulfo-
vibrio species were investigated in some research studies for their
ability to catalyse H2 formation from polarised graphite electrodes
and their ample information on electron uptake mechanisms. G.
sulfurreducens is a well-known biocatalyst that was tested in both
anode and cathode of MFCs, and also as a biocatalyst in the MEC
anode. From its four encoded hydrogenases, two are believed to be
in the cytoplasm and two others are membrane-bound hydroge-
nases involved in hydrogen uptake by this biocatalyst. Both bioa-
node inversed biocathode of a two-chambered MEC and biocathode
origin of a single-chambered MEC inoculated with this biocatalyst
displayed a considerable level of hydrogen production at the cathode
31
potential of less than  0.9 V [32]. As previously mentioned in this
section, energy-conserving hydrogenases, or cytoplasmic hydroge-
nases coupled with membrane-bound ATPase, are believed to be
energy conservation-associated mechanisms for hydrogen produc-
tion in biocathodes. G. sulfurreducens illustrated the presence of two
cytoplasmic hydrogenases, yet no possession of energy-conserving
hydrogenases were recorded by this genome [59,65]. Geelhoed et al.
reported that the proton gradient resulted by these hydrogenases
through the cytoplasmic membrane could be controlled by a
membrane-bound ATPase [32]. Furthermore, the cell attachment
process on the anode and the cathode of the single-chambered
setup also required additional study. It was assumed that the cell
may detach after growth on the bioanode and initiate its function on
the cathode. Desulfovibrio species are known microorganisms for
their ability to catalyse hydrogen production reaction [58,66].
Attached Desulfovibrio. paquesii to the graphite biocathode could
successfully catalyse hydrogen generation at the cathode potential of
less than  900 mV with 100% columbic efficiency [34]. Croese et al.
investigated the microbial community in a biocathode MEC inocu-
lated with a mixed culture. Proteobacteria and Firmicutes phyla were
identified as the dominant bacterial population, and Desulfovibrio
vulgaris as the dominant species. Subsequently, the application of
Desulfovibrio G11 as the pure culture of biocathode MEC was
investigated at the outset in the same study, without any mediator,
at cathode potential of 0.7 V [33]. Calculating the energy available
at the cathode by considering the cathode potential ( 0.7 V)
required energy for hydrogen production ( 0.41 V) and overpoten-
tial concentration since cathode loss displayed enough available
energy for microorganism growth. While some researchers now
recognise the dominant species in biocathode MEC, further studies
are required regarding biocathodes developed with pure cultures of
dominant species in order to examine the involved genes in
reduction half reactions. In addition, increased deliberation on the
growth rate and yield, as well as, energy conservation and EET
mechanism for biocathodic reaction are further necessary. An over-
view of the bacterial communities characterised in various biocata-
lysed cathode systems, along with the carbon source, detected
products and respective analysis methods were collectively tabu-
lated in Table 3.
5. Future work
Biocathode MEC is a new technology that could play a significant
role in leading recent attempts towards the delivery of bioeletro-
chemical systems out of the lab scale and into practical implementa-
tion by the replacement of precious metal catalysts. In this way,
fundamental studies are necessary to characterise effective para-
meters, individually and integrally, to improve process yield in the
form of energy and product. Additional research is mandatory to
improve the performance of biocathode in MECs with respect to the
hydrogen production rate and required applied voltage, which is still
in turn, lower and higher than those achieved by metal-based
cathodes [26,67–69]. Comprehensive studies are required to focus
on the investigation and enrichment procedures of bacterial commu-
nities as the main catalysts in the cathode chamber to improve
hydrogen formation. Although mixed cultures may consume a wider
range of sources and are easier for maintenance, fundamental
research on identified species of mixed cultures capable of providing
hydrogen is still necessary to clarify the electron uptake mechanism.
In spite of the many discussions on assumptions and hypotheses
regarding the theoretical electron uptake mechanism from the
cathode, a lack of experimental research is quite apparent in this
area. Furthermore, a more comprehensive understanding of the
reaction mechanism involved in the biocathode will aid in energy
management, whether used by the biocathode for product formation
32 T. Jafary et al. / Renewable and Sustainable Energy Reviews 47 (2015) 23–33
purposes or consumed by microorganisms for growth. The link of
hydrogen production to energy conservation and biocatalyst growth
remains unclear. The electron transfer mechanism that uses the
cathode as the electron donor, also needs to be understood as well.
Further investigation is also necessary on the energy conservation in
the biocathode of the MEC alongside studies on growth rate, yield and
kinetic [32,33,50].
6. Conclusion
Efficient hydrogen production in the biocathode MEC is a promising
goal that can assist this novel technology to overwhelm the aspects of
cost associated to current chemical catalyst sources application. The
highest hydrogen production of 2.2 m3 H2/m3/day was achieved in a
half biological two-chambered MEC with acetate and mixed microbial
community in comparison to the first established biocathode MEC with
0.63 m3 H2/m3/day. Desulfovibrio species are recognised among
detected gens in enriched mixed microbial communities capable of
producing hydrogen when attached to an electrode surface. While the
half biological setup displayed higher cathodic recovery and hydrogen
product, it is nevertheless susceptible to high hydrogen loss across the
membrane. On the other hand, it was reported that hydrogen loss
across the membrane may be less important when high current
density flows through the circuit. Although full biological two-
chambered setup may produce high current density, it nevertheless
suffers from other problems such as the precipitation of electrodes and
side product formation. Single-chambered MEC does not suffer from
problems of resistance, scaling and product loss associated with
existing membrane. However, it still suffers from very low product
yield and contamination. Furthermore, the reactor design such as size
and cathodic flow pattern had also demonstrated considerable effects
on the microbial community of the biocathode biofilm which is one of
the most interesting and unknown aspect of the field. Biocatalyst type,
enrichment process and nutrient medium are other factors that affect
the product evolution yield. Using acetate as the carbon source in the
biocathode chamber resulted in the highest hydrogen production rate
and had significantly decreased the start-up time. However, methane
detection still led some researchers to use bicarbonate and carbon
limiting strategies to overcome the methanogenesis growth. The long
start-up duration could be overcome by using acetate as the carbon
source to produce pure hydrogen product, in spite of the reported
carbon limiting strategy due to methane detection. Energy manage-
ment and the associated mechanism with electron–electrode–hydro-
gen evolution are still under investigation by scientists. Experimental
discovery on the electron transfer mechanisms may further assist the
enhancement in the present low hydrogen rate and cathode hydrogen
recovery of biocatalysed cathode MECs, in comparison to metal based
cathode MECs.
Acknowledgment
The authors gratefully acknowledge support given for this work
by 03-01-02-SF0985 Sciencefund from Malaysian Ministry of
Science, Technology & Innovation, DIP-2012-27 from Dana Impak
Penebitan from Universiti Kebangsaan Malaysia and ERGS/1/2012/
TK05/UKM/01/2 Exploratory Research Grant Scheme from Malay-
sian Ministry of Education. The author would like to thank
Hexagon Synergy (M) Sdn Bhd for the support as well.
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