Enzyme and Microbial Technology 47 (2010) 179–188

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Enzyme and Microbial Technology

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

Review

Biocatalysts in microbial fuel cells

Vinay Sharma, P.P. Kundu ∗
Department of Polymer Science & Technology, University of Calcutta, 92, A. P. C. Road, Kolkata-700009, India

a r t i c l e i n f o a b s t r a c t

Article history: The advent behind microbial fuel cells (MFC) is to provide clean electricity from the waste organic mate-
Received 31 July 2009 rial. The MFC produces electricity with the help of microorganisms. In the present review, the biocatalysts
Received in revised form 30 June 2010 or microorganisms used in the MFCs are discussed. The most used microorganisms in the MFCs belong
Accepted 2 July 2010
to Shewanella, Proteobactor and Pseudomonas families. In waste water based MFCs, mixed cultures are
mostly used. This review covers the biocatalysts used in both anode and cathode. In the recent times,
Keywords:
one of the most valuable development in the MFCs is the use of biocathodes, which eliminated various
MFC
drawbacks of these cells and enhanced the power generation capabilities as well as the production of
Biocatalyst
Shewanella
some useful gases like hydrogen. The present state of art of this technology still requires development in
Pseudomonas certain power output areas such as improvement of efficiency and cost reduction.
Geobacter © 2010 Elsevier Inc. All rights reserved.
Wastewater species
Biocathode

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179
1.1. Microbial fuel cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179
2. Bacteria used in MFC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180
2.1. Shewanella species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180
2.2. Pseudomonas species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 181
2.3. Geobacter species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 181
2.4. Wastewater species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
2.5. Biocathodes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185
2.6. Miscellaneous species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
3. Conclusions and future prospects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187

1. Introduction 1.1. Microbial fuel cells

Fuel cells need not be big to sit outside buildings or in rooms, nor The microbial fuel cell is a device that converts biochemical
need they be streamlined to fit in an automobile. They can be very energy into electrical energy with the aid of the catalytic reaction of
small and can run on organic matter. There is presently a niche mar- microorganisms [1]. Microbial fuel cells use bacteria or yeast cells
ket for stationary fuel cells, about the size of a one-car garage with as a catalyst, direct electron transport or in some cases mediators
300–200 kW fuel-cell power plants operating around the world. as electron shuttles and oxidizing agents as electron acceptors. The
Biological waste materials may be anaerobically digested, sub- structure of the fuel cell is essentially an anode compartment with
jected to pyrolysis, or otherwise pretreated to release hydrogen. cells, with or without mediator, and an electrode separated from
Unfortunately, raw biobased materials cannot be thrown into a fuel a cathode compartment. Membranes can be required if the anode
cell, because they currently require preliminary processing. chamber is anaerobic and the cathode uses oxygen. The cathode
compartment comprises an electrode and an electron acceptor. The
anode and cathode are connected via a circuit and electrons flow
∗ Corresponding author. Tel.: +91 33 23525106; fax: +91 33 23525106. from the biological cells to the cathode electron acceptor because
E-mail address: ppk923@yahoo.com (P.P. Kundu). E◦ potentials of the active components are arranged in an ascend-

0141-0229/$ – see front matter © 2010 Elsevier Inc. All rights reserved.
doi:10.1016/j.enzmictec.2010.07.001
180 V. Sharma, P.P. Kundu / Enzyme and Microbial Technology 47 (2010) 179–188

Table 1
Species studied by the researchers in anode chamber.

S. no. Species References

1. E. coli Potter [14], Zhang et al. [15],

2. Shewanella oneidensis DSP10
3. Shewanella oneidensis MR-1
4. Shewanella putrefaciens
5. Pseudomonas aeruginosa
6. Geobacter sulfurreducens
7. Geobacteraceae
Fig. 1. Schematic of the basic components of a microbial fuel cell. The anode and
8. Geobacter metallireducens
cathode chambers are separated by a membrane. The bacteria grow on the anode,
9. Dessulfobulbus propionicus
oxidizing organic matter and releasing electrons to the anode and protons to the
10. Geothrix fermentans
solution. The cathode is sparged with air to provide dissolved oxygen for the reac-
11. Paracoccus denitrificans and
tions of electrons, protons and oxygen at the cathode, with a wire (and load)
Paracoccus pantotrophus
completing the circuit and producing power. The system is shown with a resistor
12. Rhodopseudomonas palustris
used as the load for the power being generated, with the current determined based
DX-1
on measuring the voltage drop across the resistor using a multimeter hooked up to
13. Klebsiella pneumoniae
a data acquisition system.
Habermann and Pommer [22],
Zou et al. [59], Park and Zeikus
[60], Qiao et al. [61], Xi and Sun
[62]
Ringeisen et al. [16], Biffinger
et al. [18,19]
Manohar et al. [17], Biffinger et
al. [18]
Kim et al. [1], Park and Zeikus
[21]
Habermann and Pommer [22],
Rabaey et al. [23–24]
Bond et al. [26], Reguera et al.
[27,31], Trinh et al. [33]
Holmes et al. [29], Bond et al.
[30]
Min et al. [32]
Lovley et al. [53]
Lovley et al. [54]
Rabaey et al. [55]
Xing et al. [56]
Lewandowski et al. [57,58]

ing order. A schematic of an MFC system is shown in Fig. 1. Almost
all reported microbial fuel cells employ bacteria as the catalyst and
the trend is to use a consortium of organisms, often isolated from
the waste stream, and then subjected to selection in the fuel-cell
environment. Power produced is very low, but techniques exist to
convert this to useful power levels. Very dilute organic wastes that
cannot serve as substrates in other energy production systems can
be used as energy source for microbial fuel cells. The microbial fuel
cell not only recovers energy from dilute waste, but will also simul-
taneously bioremediate the waste, a process that currently requires
energy input [2,3].
The MFC that utilizes mediator as electron shuttle is called
mediator-based-MFC. The electron transfer from certain microbial
cells to the electrode is facilitated by the help of mediators such as
thionine, methylene blue, humic acid and so on [4]. Electrons are
captured by the oxidized mediator and transferred to the anode.
At the anode the mediator is oxidized, a process that releases elec-
trons to the anode and returns the mediator to its reduced state.
The ideal mediator has the following properties: (i) It should dis-
play reversible redox reaction to function as an electron shuttle;
(ii) It should have appreciable solubility in an aqueous solution and
stability; (iii) It should facilitate the electron transfer; and (iv) It
should have low formal potential. The lower the formal potential,
the larger the cell voltage since it is the difference between the cath-
ode and anode potentials [5]. Up to now most study have focused
on the generation of electricity by Fe(III)-reducing bacteria [1,6],
glucose and starch fermenting bacteria [7], Sulfate-reducing bac-
teria [8] and others such as E. coli, Enterobacter aerogens and so on
[4]. The detailed list of different microorganism at anode is given
in Table 1.
However, there are MFCs in which mediator is excluded, known
as mediatorless-MFC. In the mediatorless-MFC, electrochemically
active bacteria are used to ensure high rates of fuel oxidation and
electron transfer for the production of electrical energy.
2. Bacteria used in MFC
The development of processes that can use bacteria to produce
electricity represents a plausible method for bioenergy produc-
tion as the bacteria are self-replicating, and thus the catalysts for
organic matter oxidation are self-sustaining. Bacterial reactions can
be carried out over a wide range of temperatures depending on
the tolerance of bacteria, ranging from moderate or room-level
temperatures (15–35 ◦ C) to both high temperatures (50–60 ◦ C) tol-
erated by thermophiles [9] and low temperatures (<15 ◦ C), where
psychrophiles [10] can grow. Virtually, any biodegradable organic
matter can be used in a MFC, including volatile acids, carbohydrates,
proteins, alcohols, and even relatively recalcitrant materials like
cellulose [11–13,2,3]. Chemical mediators or electron shuttles were
routinely added to MFCs that resulted in electron transfer by bacte-
ria and even yeast [14]. In the earliest studies by Potter [14] in 1911,
the yeast Saccharomyces cerevisae and bacteria such as Bacillus coli
(later classified as Escherichia coli or E. coli) were shown to produce
a voltage, resulting in electricity generation. How that worked was
not well known as there were no known mediators added to the
cell suspensions, and E. coli [15] and yeast were not known to pro-
duce electricity at that time in the absence of mediators. Some of
the bacterial species employed in MFCs are described as below.
2.1. Shewanella species
Shewanella putrefaciens is a Gram-negative marine bacterium.
S. putrefaciens is also a facultative anaerobe with the ability to
reduce iron and manganese metabolically; i.e., it can use iron and
manganese as the terminal electron acceptor in the electron trans-
port chain (in contrast to obligate aerobes which must use oxygen
for this purpose). It is also one of the organisms associated with
the odor of rotting fish, as it is a marine organism which pro-
duces trimethylamines (hence, the species name putrefaciens, from
putrid). In both solid and liquid media, S. putrefaciens is often rec-
ognizable by its bright pink color. On solid media, the colonies are
round, fast-growing, and pink. The organism is also fast-growing in
liquid media.
Ringeisen et al. [16] used Shewanella oneidensis DSP10 in growth
medium with lactate and buffered ferricyanide solutions as anolyte
and catholyte, respectively. Maximum power densities of 24 and
10 mW/m2 were measured using the true surface areas of retic-
ulated vitreous carbon (RVC) and graphite felt (GF) electrodes
without the addition of exogenous mediators in the anolyte. Cur-
rent densities at maximum power were measured as 44 and
 V. Sharma, P.P. Kundu / Enzyme and Microbial Technology 47 (2010) 179–188
20 mA/m2 for RVC and GF, while short circuit current densities
reached 32 mA/m2 for GF anodes and 100 mA/m2 for RVC. The addi-
tion of electron mediators resulted in current and power increases
of 30–100%. These power densities were surprisingly high for a pure
S. oneidensis culture. They observed that the short diffusion lengths
and the high ratio of surface area to chamber volume utilized in the
mini-MFC, enhanced the power density when compared to output
from similar macroscopic MFCs.
Manohar et al. [17] determined the internal resistance (Rint ) of a
mediatorless microbial fuel cell (MFC) as a function of cell voltage
using electrochemical impedance spectroscopy (EIS) for a MFC with
and without S. oneidensis MR-1. The same tests were performed
for a MFC containing small stainless steel (SS) balls in the anode
compartment with a graphite feeder electrode in a packed bed cell.
They observed that Rint decreased with decreasing cell voltage as
the increasing current flow decreased the polarization resistance
of the anode and the cathode. In the presence of MR-1, Rint was
lower by a factor of about 100 than Rint of the MFC with buffer and
lactate as anolyte. Rint was also significantly lower for the anode
containing SS balls with buffer and lactate as anolyte. For the MFC
containing SS balls in the anode compartment, further no signifi-
cant decrease of Rint was obtained. When MR-1 was added to the
anolyte, the polarization resistance of the anode was lower than
that of the cathode.
Biffinger et al. [18] compared S. oneidensis MR-1 with DSP 10.
Miniature MFCs using bare graphite felt electrodes and nanoporous
polycarbonate membranes with MR-1 or DSP10 cultures generated
>8 W/m3 and ∼400 ␮A between pH 6–7. The DSP10 strain signifi-
cantly outperformed MR-1 at neutral pH. Higher concentrations of
DSP10 were sustained at pH 7 relative to that of MR-1. Whereas
at pH 5, this trend was reversed, indicating that the cell count was
perhaps not solely responsible for the observed differences in cur-
rent, but also by changes in the amount of autologous redox active
mediators in the growth media at lower pH values. S. oneidensis
MR-1 was determined to be more suitable than DSP10 for MFCs
with elevated acidity levels. The concentration of riboflavin in the
bacterial cultures was reduced significantly at pH 5 for DSP10, as
determined by high performance liquid chromatography (HPLC) of
the filtered and sterilized growth media. In addition, these results
suggest that mediator biosynthesis and not solely bacterial concen-
tration plays a significant role in current output from S. oneidensis
containing MFCs.
In another report, Biffinger et al. [19] used a miniature-microbial
fuel cell to monitor biofilm development from a pure culture of
S. oneidensis DSP10 on graphite felt (GF) under minimal nutri-
ent conditions. They observed the formation of biofilms after 5
days at 25 ◦ C. The power generated with the biofilm-enhanced
anode (26 W/m3 for Pt/C GF–O2 and 250 W/m3 for uncoated
GF-ferricyanide) was 60% of that generated with the planktonic
culture (150 W/m3 for Pt/C GF–O2 and 420 W/m3 for uncoated GF-
ferricyanide). The power output increased during the operation of
the fuel cell as the solution-phase bacterial concentration increased
in the reservoir. The power densities per unit volume produced by
the DSP10-only biofilm was comparable with MFCs using mixed
cultures with either ferricyanide or oxygen reduction cathodes.
Kim et al. [1] used S. putrefaciens in a mediatorless-MFC. They
studied direct electron transfer from different S. putrefaciens strains
to an electrode through cyclic voltammetry and a fuel-cell type
electrochemical cell. Both methods studied the electrochemical
activity of the bacterium without any electrochemical mediators.
It was observed that the anaerobically grown cells of S. putre-
faciens MR-1, IR-1, and SR-21 showed electrochemical activities,
but no activities were observed in aerobically grown S. putre-
faciens cells. The current generation from the metal reducing
bacterium depended on the electrochemical activity of bacterium,
principally oxidation of fuel by the bacterial metabolism, and
181
the direct electron transfer capacity from the bacterial surface to
the electrode.
In another study, cysteine was used as substrate for the elec-
tricity generation using a MFC [20]. 16S ribosomal RNA (16S rRNA)
based analysis of the biofilm on the anode of the MFC indicated
that the predominant organisms were Shewanella spp. closely
related to Shewanella affinis. It was reported that over a period of
a few weeks, electricity generation gradually increased to a maxi-
mum power density of 19 mW/m2 (1000  resistor; 385 mg/L of
cysteine). Power output increased to 39 mW/m2 , when cysteine
concentrations were increased up to 770 mg/L and external resistor
changed to 493 . The use of a more active cathode with Pt or Pt–Ru,
increased the maximum power from 19 to 33 mW/m2 , showing
that the cathode efficiency had limited the power generation. Elec-
tron recovery was 14% based on complete cysteine oxidation, with
an additional 14% (28% total) potentially lost to oxygen diffusion
through the proton exchange membrane. Park and Zeikus [21] stud-
ied the effect of electrode composition on the electricity generation
in a single compartment fuel cell using S. putrefaciens. They reported
that the electricity production was dependent on anode compo-
sition, electron donor type and cell concentration. A maximum
current of 2.5 mA and a current density of 10.2 mW/m2 electrode
were obtained with a Mn4+ graphite anode, 200 mM sodium lactate
and a cell concentration of 3.9 g cell protein/mL. Current production
by S. putrefaciens was enhanced 10-fold when an electron mediator
(i.e., Mn4+ or neutral red) was incorporated into the graphite anode.
2.2. Pseudomonas species
Pseudomonas aeruginosa is a member of the Gamma Proteobac-
teria class of Bacteria. It is a Gram-negative, aerobic rod belonging
to the bacterial family Pseudomonadaceae. Since, the revisionist
taxonomy is based on conserved macromolecules (e.g. 16S rRNA)
the family includes only members of the genus Pseudomonas. P.
aeruginosa is the type species of its group, which contains 12 other
members. Habermann and Pommer [22] in 1991 studied biologi-
cal fuel cells using cost effective fuel storing materials like sulfide.
They pioneered a low maintenance fuel-cell system with long-
term stability and high short-term output. They used a variety of
microorganisms. The various bacterial strains were P. aeruginosa, E.
coli, Proteus vulgaris, etc.
Rabaey et al. demonstrated that exogenous mediators did
not have to be added to a culture [23,24]. These self-produced
or endogenous chemical mediators, for example pyocyanin and
related compounds produced by P. aeruginosa, can shuttle elec-
trons to an electrode and produce electricity in an MFC [24]. The
production of high concentrations of mediators by mixed cultures
primarily containing P. aeruginosa, coupled with a very low internal
resistance MFC were achieved by using ferricyanide as a catholyte
(instead of oxygen), produced 3.1–4.2 W/m2 in MFCs [24]. When
the substrate was exhausted, soluble mediators were removed,
making it difficult for these compounds to accumulate to high con-
centrations.
2.3. Geobacter species
Geobacter [25] is a genus of proteobacteria. Geobacter are an
anaerobic respiration bacterial species which have capabilities
that may make them useful in bioremediation. The geobacter was
found to be the first organism with the ability to oxidize organic
compounds and metals, including iron, radioactive metals and
petroleum compounds into environmentally benign carbon diox-
ide while using iron oxide or other available metals as electron
acceptor.
Bond et al. [26] used Geobacter sulfurreducens in MFC cham-
bers, in which a graphite electrode served as the sole electron
182 V. Sharma, P.P. Kundu / Enzyme and Microbial Technology 47 (2010) 179–188
acceptor and acetate or hydrogen was the electron donor. The
electron-accepting electrodes were maintained at oxidizing poten-
tials by connecting them to similar electrodes in oxygenated
medium (fuel cells) or to potentiostats that poised electrodes at
+0.2 V versus an Ag/AgCl reference electrode (poised potential).
When a small inoculum of G. sulfurreducens was introduced into
electrode-containing chambers, the electrical current production
was dependent upon the oxidation of acetate to carbon dioxide
and increased exponentially, indicating for the first time that elec-
trode reduction supported the growth of this organism. When the
medium was replaced with an anaerobic buffer lacking nutrients
required for growth, but consisting of acetate, current production
was unaffected and acetate-dependent cells attached to these elec-
trodes continued to generate electrical current for weeks.
This represents the first report of microbial electricity produc-
tion solely by cells attached to an electrode. Electrode-attached
cells completely oxidized acetate to levels below detection
(<10 ␮M), and hydrogen was metabolized to a threshold of 3 Pa.
The rates of electron transfer to electrodes (0.21–1.2 ␮mol of elec-
trons/mg of protein/min) were similar to those observed for respi-
ration with Fe(III) citrate as the electron acceptor (E◦ = +0.37 V). The
production of current in microbial fuel cell (65 mA/m2 of electrode
surface) or poised potential (163–1143 mA/m2 ) mode was greater
than what was reported for other microbial fuel cells employ-
ing higher cell densities and electron-shuttling compounds. Since
acetate was completely oxidized, the efficiency of conversion of
organic electron donor to electricity was significantly higher than
in previously described microbial fuel cells. These results suggest
that the effectiveness of microbial fuel cells can be increased with
organisms such as G. sulfurreducens that can attach to electrodes
and remain viable for long periods of time, while completely oxi-
dizing organic substrates with quantitative transfer of electrons to
an electrode, as long as they are compatible with other species in a
mixed culture.
Reguera et al. [27] studied extracellular electron transfer
through microbial nanowires. In this MFC, pili of G. sulfurreducens
served as biological nanowires, transferring electrons from the cell
surface to the surface of Fe(III) oxides. Electron transfer through pili
indicated possibilities for other unique cell–surface and cell–cell
interactions, and for bioengineering of novel conductive materials.
The potential role of outer membrane proteins in electron transfer
to insoluble Fe(III) oxides by G. sulfurreducens was also investi-
gated by Mehta et al. [28]. They used two of the most abundant
proteins from the outer surfaces of intact c-type cytochrome cells.
One, designated OmcS, had a molecular mass of ca. 50 kDa and was
predicted to be an outer membrane hexaheme c-type cytochrome.
Transcripts for omcS were detected during the growth on Fe(III)
oxide, but not on soluble Fe(III) citrate. The omcS mRNA consisted
primarily of a monocistronic transcript, and to a lesser extent, a
longer transcript that also contained the downstream gene omcT,
which was predicted to encode a second hexaheme outer mem-
brane cytochrome with 62.6% amino acid sequence, an identity to
OmcS. The other abundant c-type cytochrome sheared from the
outer surface of G. sulfurreducens, designated OmcE, had a molecu-
lar mass of ca. 30 kDa and was predicted to be an outer membrane
tetraheme c-type cytochrome.
When either omcS or omcE was deleted, G. sulfurreducens was
not able to reduce Fe(III) oxide, but still reduced soluble electron
acceptors, including Fe(III) citrate. The mutants reduced Fe(III) in
Fe(III) oxide medium only, if the Fe(III) chelator, nitrilotriacetic acid,
or the electron shuttle, anthraquinone 2,6-disulfonate, was added.
Expressing omcS or omcE in the trans form restored the capacity for
Fe(III) oxide reduction. OmcT was not detected among the sheared
proteins, and genetic studies indicated that G. sulfurreducens was
not able to reduce Fe(III) oxide, when omcT was expressed. In
contrast, Fe(III) oxide was reduced, when omcS was expressed
in the absence of OmcT. These results suggested that OmcS and
OmcE were involved in electron transfer to Fe(III) oxides in G. sul-
furreducens. They also emphasized the importance of evaluating
mechanisms for Fe(III) reduction with environmentally relevant
Fe(III) oxide, rather than the more commonly utilized Fe(III) citrate.
Holmes et al. [29] investigated the potential role of Geobac-
teraceae, Geopsychrobacter electrodiphilus gen. nov., sp. nov., in
electricity production by a marine sediment fuel cell. Microorgan-
isms from the anode surface of a marine sediment fuel cell were
enriched and isolated with Fe(III) oxide. Two unique marine iso-
lates were recovered, strains A1T and A2. They are gram-negative,
non-motile rods, with abundant c-type cytochromes. Phylogenetic
analysis of the 16S rRNA, recA, gyrB, fusA, rpoB, and nifD genes indi-
cated that strains A1T and A2 represented a unique phylogenetic
cluster within the Geobacteraceae. Both strains were able to grow
with an electrode serving as the sole electron acceptor and trans-
ferred ca. 90% of the electrons available in their organic electron
donors to the electrodes. These organisms were the first psychro-
tolerant members of the Geobacteraceae reported and grown at
temperatures between 4 and 30 ◦ C, with an optimum temperature
of 22 ◦ C. Strains A1T and A2 utilized a wide range of traditional
electron acceptors, including all forms of soluble and insoluble
Fe(III) tested, anthraquinone 2,6-disulfonate, and S◦ . In addition to
acetate, both strains utilized a number of other organic acids, amino
acids, long-chain fatty acids, and aromatic compounds to support
growth with Fe(III) nitrilotriacetic acid as an electron acceptor. The
metabolism of these organisms differed in that only strain A1T
used acetoin, ethanol, and hydrogen as electron donors, whereas
only strain A2 used lactate, propionate, and butyrate. The name
G. electrodiphilus gen. nov., sp. nov., was proposed for strains A1T .
Strains A1T and A2 (ATCC BAA-770; JCM 12470) represented the
first organisms recovered from anodes that effectively coupled the
oxidation of organic compounds to an electrode.
Marine sediment microorganisms were also studied by Bond et
al. to harvest energy [30]. Energy in the form of electricity was har-
vested from marine sediments by placing a graphite electrode (the
anode) in the anoxic zone and connecting it to a graphite cathode
in the overlying aerobic water. A specific enrichment of microor-
ganisms of the family Geobacteraceae on energy harvesting anode,
showed that these microorganisms conserved energy to support
their growth by oxidizing organic compounds with an electrode
serving as the sole electron acceptor. This was useful in promoting
the bioremediation of organic contaminants in subsurface environ-
ments. The highly structured, multilayered biofilms on the anode
surface of a microbial fuel cell developed by G. sulfurreducens was
studied by Reguera et al. [31]. Cells at a distance from the anode
remained viable, and there was no decrease in the efficiency of cur-
rent production as the thickness of the biofilm increased. Genetic
studies demonstrated that efficient electron transfer through the
biofilm required the presence of electrically conductive pili. These
pili represented an electronic network permeating the biofilm that
promoted long-range electrical transfer in an energy-efficient man-
ner, increasing electricity production more than 10-fold.
Min et al. [32] compared the power outputs of the MFCs based
on a pure culture (Geobacter metallireducens) and a mixed culture
(wastewater inoculum). Power output with either inoculum was
essentially the same, with 40 ± 1 mW/m2 for G. metallireducens and
38 ± 1 mW/m2 for the wastewater inoculum. In both systems, it was
observed that oxygen diffusion from the cathode chamber into the
anode chamber was a negative factor in power generation. A critical
factor in the power density achieved in a two-chambered system
was the system internal resistance, which was primarily a function
of the proton exchange system. Trinh et al. [33] studied the electric-
ity generation with acetate as the fuel and G. sulfurreducens as the
biocatalyst on the anode electrode. They obtained a current pro-
duction of 0.20–0.24 mA at 30-32 ◦ C. The maximum power density
 V. Sharma, P.P. Kundu / Enzyme and Microbial Technology 47 (2010) 179–188
of 418–470 mW/m2 , obtained at an external resistor of 1000 , was
increased over 2-fold (from 418 to 866 mW/m2 ) as the Pt loading
on the cathode electrode was increased from 0.5 to 3.0 mg Pt/cm2 .
The optimal batch mode temperature was between 30 and 32 ◦ C
with a maximum power density of 418–470 mW/m2 . It was also
noted that the power density increase was disproportional to the
platinum concentration increase.
2.4. Wastewater species
Habermann and Pommer [22] first reported the wastewater
based MFCs in 1991. Logan and co-workers [34,35] have done
considerable work on the electricity generation from wastewater
microorganisms. Min and Logan [34] also used a flat plate micro-
bial fuel cell (FPMFC) for continuous electricity generation from
domestic wastewater and organic substrates. This microbial fuel
cell was designed to function as plug flow reactor using a combined
electrode/proton exchange membrane (PEM) system. The reactor
consisted of a single channel formed between two nonconductive
plates that were separated into two halves by the electrode/PEM
assembly also known as membrane electrode assembly (MEA). Each
electrode was placed on an opposite side of the PEM, with the anode
facing the chamber containing the liquid phase and the cathode fac-
ing a chamber containing only air. Electricity generation using the
FPMFC was examined by continuously feeding a solution contain-
ing wastewater, or a specific substrate, into the anode chamber.
The system was initially acclimated for 1 month using domestic
wastewater or wastewater enriched with a specific substrate such
as acetate. Average power density using only domestic wastewater
was 72 ± 1 mW/m2 at a liquid flow rate of 0.39 mL/min [42% COD
(chemical oxygen demand) removal, 1.1 h HRT (hydraulic reten-
tion time)]. At a longer HRT = 4.0 h, there was 79% COD removal
and an average power density of 43 ± 1 mW/m2 . Power output
was found to be a function of wastewater strength according
to a Monod-type relationship, with a half-saturation constant of
Ks = 461 or 719 mg COD/L. Power generation was sustained at high
rates with several organic substrates (all at ∼1000 mg COD/L),
including glucose (212 ± 2 mW/m2 ), acetate (286 ± 3 mW/m2 ),
butyrate (220 ± 1 mW/m2 ), dextran (150 ± 1 mW/m2 ), and starch
(242 ± 3 mW/m2 ). These results demonstrated the versatility of
power generation in a MFC with a variety of organic substrates and
showed that power can be generated at a high rate in a continuous
flow reactor system.
The effect of electrode spacing and advective flow through the
porous anode on the power generation by a microbial fuel cell has
been studied by Logan and co-workers [35]. It was observed that the
maximum power density from a MFC with glucose decreased from
811 to 423 mW/m2 , when the electrode spacing was decreased
from 2 to 1 cm. The MFC configurations showing different elec-
trode spacing are shown in Fig. 2. When the electrode spacing
was reduced from 2 cm (anode exposed to only one side of the
fluid; case A in Fig. 2) to 1 cm (anode exposed to both sides of the
fluid; case B), the maximum power density with glucose (500 mg/L)
decreased from 811 to 684 mW/m2 . If the anode was placed 1 cm
from the cathode and exposed to only one side of the fluid (case
C), the maximum power density further decreased to 423 mW/m2 .
Power density decreased under these conditions even though Rint
decreased from 35 (2 cm) to 16  (1 cm) (cases A and C). The
decrease in Rint should have resulted in an increase in power output,
even though the difference in Rint was insignificant. The difference
was insignificant due to the reported Rint being at the wrong level
of magnitude, since with EIS it can only represent the ohmic losses.
Still, it was very interesting to see that power density also decreased
with the improvement in the internal losses. The coulombic effi-
ciency (CE) decreased from 28% (A) to 18% (C) with a decrease in
the electrode spacing. The decrease in the power density in both
183
cases, when the anode was moved closer to the cathode, resulted
from decreased activity of the bacteria on the anode, as shown
by a decrease in the open circuit potentials (OCPs) of the anode.
The open circuit voltages (OCVs) measured for these three cases
were 0.820 V (A), 0.797 V (B), and 0.783 V (C). The OCPs of the cath-
ode were essentially constant (0.268 V, A; 0.267 V, B; and 0.266 V,
C). Thus, the changes in the OCVs were directly a result of the
increased anode potentials, which were −0.552 V (A), −0.531 V (B),
and −0.516 V (C).
However, providing advective flow through the porous anode
toward the cathode substantially increased the power density,
resulting in the highest maximum power densities yet achieved
in an air-cathode system using glucose or domestic wastewater
as substrates (Fig. 2D). For glucose, with a 1-cm electrode spac-
ing and flow through the anode with continuous flow operation of
the MFC, the maximum power density increased to 1540 mW/m2
(51 W/m3 ) and the CE increased to 60%. Using domestic wastew-
ater (255 ± 10 mg of COD/L), the maximum power density was
464 mW/m2 (15.5 W/m3 ; CE = 27%), even though the flow through
the anode could lead to plugging, especially for particulated sub-
strates such as the domestic wastewater. Still, the system was
operated using glucose for over 42 days without clogging. These
results showed that power output in the air-cathode single-
chamber MFC can be increased by reducing the electrode spacing, if
the reactors operate in continuous flow mode with advective flow
through the anode toward the cathode.
Ieropoulos et al. [36] also referred to the inefficiency of the
large-scale reactor. They studied MFC base on carbon veil and inves-
tigated the effect of stack configuration and scalability. In terms of
power density expressed as per unit of electrode surface area and
as per unit of anode volume, the small-sized MFC was superior to
both the medium- and large-scale MFCs by a factor of 1.5 and 3.5,
respectively. Based on measured power output from 10 small units,
a theoretical projection for 80 small units (giving the same equiv-
alent anodic volume as one large 500 mL unit) gave a projected
output of 10 W/m3 , which was approximately 50 times higher than
the recorded output produced by the large MFC.
Ieropoulos et al. [37] investigated the behavior of MFC in water
by linking the cathodic half cell to an artificial gill. The current
output at ambient temperature was 32 ␮A, which was noted to
increase ∼100 ␮A (200%) at 52 ◦ C. Also the increase in water flow
rate resulted in an increase in the output ranging from 135% to
150%. Liu et al. [38] studied the effect of reactor architecture in
membrane-free single-chamber microbial fuel cells. The maximum
power density generated by the larger MFC (LMFC) containing a
cloth electrode was 16 W/m3 (520 mW/m2 -cathode area) at a cur-
rent density of 0.18 mA/cm2 , which was slightly higher than that
(14 W/m3 ) of the smaller MFC (SMFC). They studied the effect
of anode surface area, ionic strength, anode orientation, reactor
type and biofilm on the performance of the microbial fuel cell.
They observed that the performance was improved to 20 W/m3
(630 mW/m2 ) at current density of 0.26 mA/cm2 when the ionic
strength of the solution was increased from 100 to 300 mM as a
result of the decreased internal resistance (Rint = 7.3 ).
He et al. [39] used artificial wastewater in an upflow micro-
bial fuel cell for electricity generation. The reactor was fed with
sucrose solution as the electron donor for a 5-month period and
continuously generated electricity with a maximum power den-
sity of 170 mW/m2 . The power density increased with an increasing
chemical oxygen demand (COD) loading rates up to 2.0 g COD/L/day
after which, no further increase in power density were observed,
indicating the presence of limiting factors. The internal resistance
was the main limiting factor for UMFC, which was 85  at the
maximum power density and restricted the power output by caus-
ing a significant decrease in the operating potential. Fan et al.
[40] used bicarbonate buffer in microbial fuel cell and also out-
184 V. Sharma, P.P. Kundu / Enzyme and Microbial Technology 47 (2010) 179–188

Fig. 2. MFC configurations showing different electrode spacings. MFCs operated in batch mode with (A) 2-cm spacing, (B) 1-cm spacing with anode placed within chamber,
and (C) 1-cm spacing. (D) Schematic and (E) prototype of the MFC with four sections used in continuous flow tests, where X, the distance between the electrodes (shown
with two sections, or X = 2 cm) is increased by adding additional 1-cm-long sections. The width of the first two sections (inlet chamber) is fixed at 2 cm.
Reproduced with the permission of Environ Sci Technol 2006;40:2428 ©ACS Publishing, USA.

lined the proton transfer mechanism. The performances of MFCs
with cloth electrode assemblies (CEA) were evaluated using bicar-
bonate buffer solutions. A maximum power density of 1550 W/m3
(2770 mW/m2 ) was obtained at a current density of 0.99 mA/cm2
using a pH 9 bicarbonate buffer solution. Such a power density was
38.6% higher than that using a pH 7 phosphate buffer at the same
concentration of 0.2 M. They proposed a mechanism for the proton
transfer (shown in Fig. 3) in the MFC on the basis of the quantita-
tive comparison of free proton transfer rates, diffusion rates of pH
buffer species, and the generated current.
Venkata Mohan et al. [41,42] used wastewater based MFC
for the bioelectricity production. In another report, they [41]
worked on a mediatorless-MFC in fed batch mode, using
selectively enriched hydrogen producing mixed culture under
acidophilic microenvironment. Maximum voltage of 271.5 mV
(5.43 mA) and 304 mV (6.08 mA) was recorded at operat-
ing organic loading rates (OLR) of 1.165 kg COD/m3 day and
1.404 kg COD/m3 day, respectively, when measured at 50 
external. COD removal efficiency of 35.4% {substrate degra-
dation rate (SDR) of 0.412 kg COD/m3 day} and 62.9% {SDR,
0.88 kg COD/m3 day} was observed at OLRs 1.165 kg COD/m3 day
and 1.404 kg COD/m3 day, respectively. Maximum specific power
production of 0.163 W/kg CODR (1.165 kg COD/m3 day; 50 ) and
0.198 W/kg CODR (1.404 kg COD/m3 day; 100 ) was observed
during a stable phase of fuel-cell operation. Current den-
sity of 747.96 mA/m2 (1.165 kg COD/m3 day) and 862.85 mA/m2
(1.404 kg COD/m3 day) was observed at 10 .
The effect of anodic biofilm growth on bioelectricity production
in single-chambered mediatorless-MFC using designed synthetic
wastewater (DSW) and chemical wastewater (CW) was studied
by Venkata Mohan et al. [42]. Three MFCs (plain graphite elec-
trodes, air-cathode, Nafion® membrane) were operated separately
with variable biofilm coverage [control; anode surface coverage
(ASC), 0%], partially developed biofilm [PDB; ASC ∼44%; 90 days]
and fully developed biofilm [FDB; ASC ∼96%; 180 days] under aci-
dophilic conditions (pH 6) at room temperature. The study depicted
 V. Sharma, P.P. Kundu / Enzyme and Microbial Technology 47 (2010) 179–188
Fig. 3. Diagram of proton transfer mechanism in air-cathode MFCs with a
cloth/membrane layer. The monobasic phosphate/dibasic phosphate ion-pair
accounts for the major mechanism for protons transfer in air-cathode MFCs using
phosphate buffer, while bicarbonate–carbonate is the major proton carrier for MFCs
using bicarbonate buffer.
Reproduced with the permission of Environ Sci Technol 2007;41:8156 ©ACS Pub-
lishing, USA.
the effectiveness of anodic biofilm formation in enhancing the
extracellular electron transfer in the absence of mediators. Cyclic
voltammetry analysis showed six-fold increment in energy out-
put from control (1.812 mJ) to PDB (10.666 mJ) operations and
about eight-fold increment in energy from PDB to FDB (86.856 mJ).
Biofilm configured MFC was shown to have the potential to selec-
tively support the growth of electrogenic bacteria with robust
characteristics, capable of generating higher power yields along
with substrate degradation especially operated with characteris-
tically complex wastewaters as substrates.
Huang et al. [43] studied the xylose degradation and gener-
ated electricity using a two-chamber mediatorless-MFC. Voltage
output followed saturation kinetics as a function of xylose concen-
tration for concentration below 9.7 mM, with a predicted maximum
of 86 mV (6.3 mW/m2 or 116 mW/m3 ) and half-saturation con-
stant (Ks ) of 0.29 mM. Xylose concentrations from 0.5 to 1.5 mM
resulted in coulombic efficiencies and maximum voltage ranging
from 41 ± 1.6 to 36 ± 1.2% and 55 ± 2.0 to 70 ± 3.0 mV, respectively.
Xylose degradation rate increased with increasing xylose concen-
tration up to 9.7 mM and the predicted maximum degradation rate
was 0.13 mM/h and Ks of 3.0 mM. Stirring by nitrogen in the anode
chamber led to 99 ± 2.3 mV maximum voltage (8.4 ± 0.4 mW/m2
or 153 ± 7.1 mW/m3 ) and 5.9 ± 0.3% coulombic efficiency at MFC
running time of 180 h, which were respectively 17 ± 1.2% and
37 ± 1.8%, higher than those without stirring. A better proton trans-
port through the buffer solution resulted in higher utilization of the
electron donor. The COD removal under stirring was 22.1 ± 0.3%,
which was slightly lower than that of 23.7 ± 0.4% under no stirring.
However, stirring resulted in 59% lower xylose degradation rate.
Min et al. [44] also developed a submersible MFC (SMFC) by
immersing an anode electrode and a cathode chamber in an anaero-
bic reactor. They used domestic wastewater amended with acetate
as medium. The maximum power generation was 204 mW/m2
with current density of 595 mA/m2 at a circuit resistance of 180 
(SCOD of wastewater = 1672 ± 6 mg/L). The power output showed
a saturation-type relationship as a function of wastewater concen-
tration (SCOD), with a half-saturation coefficient of Ks = 244 mg/L
and a maximum power density of Pmax = 244 mW/m2 .
2.5. Biocathodes
The conventional MFC is a two-chamber system, consisting
of anode and cathode chambers that are separated by a proton
185
exchange membrane (PEM). This system has been half biologi-
cal, because only the anode side contains electrochemically active
microorganisms, while the cathode is abiotic. For this reason some
researchers [45–50] have recently started working on the concept
of biocathodes. Instead of being catalyzed by platinum, the com-
bination of protons, electrons and oxidants at the cathode could
be catalyzed by a bacterial reaction. Biocathodes are of two types:
(i) aerobic biocathodes use oxygen as the oxidant and microorgan-
isms to assist the oxidation of transition metal compounds, such
as Mn(II) or Fe(II), for electron delivery to oxygen; (ii) anaerobic
biocathodes use compounds such as nitrate, sulfate, iron, man-
ganese, selenate, arsenate, urinate, fumarate and carbon dioxide
as terminal electron acceptors.
A baffle-chamber membraneless MFC was used for electricity
generation by Hu [45]. They studied the influence of reactor config-
urations designed to mitigate the impact of oxygen transport on the
electricity generation. The reactor was constructed to reduce mix-
ing in the vicinity of cathode and facilitate thick (>1 mm) biofilm
formation on the cathode by adding anaerobic biomass/sludge
(4330 ± 410 mgCOD/L), resulting in an overall coulombic efficiency
of more than 30% at glucose concentrations ranging from 96 to
960 mg COD/L, compared to previously reported efficiencies of
<10% in a completely mixed membraneless MFC. Efficiencies in
the absence of anaerobic sludge dropped to 21.2 ± 3.7%, suggest-
ing the importance of pH buffering provided by the biomass in
improving electron transport to the anode. However, the anaerobic
sludge itself provided a very limited power density (approximately
0.3 mW/m2 ) and power generation was primarily associated with
glucose degradation (e.g., 129 ± 15 mW/m2 ).
Lefebvre et al. [46] reported a microbial fuel cell equipped with a
biocathode for organic removal and denitrification. They designed a
two-chambered MFC for the electricity generation using a biocath-
ode. Microorganisms performed denitrification by using electrons
supplied by bacteria oxidizing domestic wastewater and acetate
as substrates in the anode chamber. This two-chambered MFC
equipped with a biocathode generated 9.4 mW/m2 of anode sur-
face or 0.19 W/m3 of anode chamber volume, while removing over
65% of COD, 84% of total nitrogen and nearly 30% of suspended solids
with domestic wastewater as a substrate. On changing from domes-
tic wastewater to sodium acetate synthetic solution, electricity
was generated immediately, which indicated the adaptability of
the electrophilic bacteria to adapt instantly to a new substrate.
Clauwaert et al. [47] utilized nitrate as electron acceptor to get
an output around 10 W/m3 . When the anode of acetate oxidiz-
ing tubular microbial fuel cell was combined with an open air
biocathode for electricity production, the maximum power pro-
duction was 83 ±11 W/m3 MFC (0.183 L MFC) for batch fed systems
(20–40% Coulombic yield) and 65 ± 5 W/m3 MFC for a continuous
system with an acetate loading rate of 1.5 kg COD/m3 day (90 ± 3%
Coulombic yield) [48]. Electrochemical precipitation of manganese
oxides on the cathodic graphite felt decreased the start-up period
with approximately 30% versus non-treated graphite felt. After the
start-up period, the cell performance was similar for the pretreated
and non-treated cathodic electrodes. Several reactor designs were
tested, and it was found that enlargement of the 0.183 L MFC reac-
tor by a factor 2.9–3.8 reduced the volumetric power output by
60–67%.
In another publication, the cell performance and microbial com-
munity was investigated using a biocathode in a MFC by Chen
et al. [49]. A two-chambered MFC with biocathode was used and
after a 2-month operation, the microorganisms of each compart-
ment (anode and cathode) were well developed and the output of
MFC increased in terms of electricity generation. At 20 mL/min flow
rate of the cathodic influent, the maximum power density reached
19.53 W/m3 , while the corresponding current and cell voltage were
15.36 mA and 223 mV at an external resistor of 14.9 , respectively.
186 V. Sharma, P.P. Kundu / Enzyme and Microbial Technology 47 (2010) 179–188

With the development of microorganisms in both compartments,

the internal resistance decreased from initial 40.2–14.0 .
Recently, Cao et al. [50] developed a photo-biocathode for
cathodic carbon dioxide reduction. The biocathode used dissolved
carbon dioxide (bicarbonate) as the acceptor. During acclimation,
the cathode was set at a potential of 0.242 V (vs. SHE) using a poten-
tiostat. After approximately 1 month of acclimation, a current of
1 mA was sustained. Bicarbonate was reduced in stoichiometric
agreement with current generation, with 0.28 ± 0.02 mol of bicar-
bonate reduced per mole of electrons. When this biocathode was
used in a two-bottle MFC, a power density of 750 mW/m2 was pro-
duced. The maximum power generated by the biocathode with
acetate as the electron donor was 750 mW/m2 . In contrast, the max-
imum power generated using a plain carbon cathode, was only
50 mW/m2 . The power density of the biocathode was therefore
15-fold larger than that achieved with an abiotic cathode control
(Fig. 4). They also compared the biocathode performance with ferri- Fig. 4. Comparison of MFC performance with abiotic control cathode, biocathode
cyanide, a commonly used chemical catholyte (50 mM K3 [Fe(CN)6 ], and ferricyanide cathode, respectively.
Reproduced with the permission of Energy Environ Sci 2009;2:500 ©RSC Publishing,
100 mM PBS, pH 7.0) (Fig. 4). Under these conditions, the maximum UK.
power generated was 1050 mW/m2 , which was 40% greater than
that of the biocathode. Results from Fig. 4 showed that the biocath-
ode performance was comparable with the chemical cathode and resistance decreased from 900 to 750 , while maximum sus-
far better than the abiotic cathode. tainable power output decreased from 32 to 28 mW/m2 . These
Aldrovandi et al. [51] investigated the power production in effects were caused by changes in the microbial ecology of anodic
a membraneless and mediatorless synthetic wastewater micro- and cathodic biomass attached to the electrodes. The untreated
bial fuel cell (shown in Fig. 5). They used a biological cathode graphite biocathode was suitable support for aerobic bacteria capa-
and a compact wastewater treatment reactor, fed with syn- ble of oxygen reduction for wastewater MFCs. This is an unexplored
thetic wastewater. When operated with an external resistance of area and requires further work.
250 , the MFC produced a long-term power of about 70 mW/m2 Simultaneous organics removal and nitrification using a novel
for 10 months. Denaturing Gradient Gel Electrophoresis (DGGE) nitrifying biocathode MFC reactor was investigated by Tran et al.
analysis of the cathode biomass, when the MFC was closed on [52]. The introduction of nitrifying biomass into the cathode cham-
a 2100  external resistance showed that the sequence bands ber caused higher voltage outputs than that of MFC operated with
were affiliated with Firmicutes, ␣-Proteobacteria, ␤-Proteobacteria, the abiotic cathode. They observed that the maximum power den-
␥-Proteobacteria, and Bacteroidetes groups. When the external resis- sity increased 18%, when the cathode was run under the biotic
tance was varied between 250 and 2100 , minimum sustainable condition and fed by nitrifying medium with alkalinity/NH4 + –N

Fig. 5. Reactor design: (1) first anaerobic chamber; (2) second anaerobic chamber; (3) aerobic chamber; (4) settling tank. The anaerobic baffled reactor (ABR, chambers
#1 and #2) sediments and ferments easily degradable COD, in series with an activated sludge process (chambers #3 and #4), which nitrifies and further reduces organic
compounds. Electrodes are connected via an external circuit on a load variable between 250 and 2100 . The volumes of the four compartments are 18, 18, 22, and 5 L,
respectively.
Reproduced with the permission of Bioresour Technol 2009;100:3253 ©Elsevier Ltd.
 V. Sharma, P.P. Kundu / Enzyme and Microbial Technology 47 (2010) 179–188
ratio of 8 (26 against 22 mW/m2 ). The voltage output was not dif-
ferentiated when NH4 + –N concentration was increased from 50 to
100 mg/L under such alkalinity/NH4 + –N ratio. The maximum power
density increased by 68% in comparison with the abiotic cath-
ode MFC (37 against 22 mW/m2 ). Polarization curves demonstrated
that the both the activation and concentration losses were lowered
during the period of nitrifying biocathode operation. Ammonium
was totally nitrified and mostly converted to nitrate in all cases
of the biotic cathode conditions, and in addition high COD removal
efficiency (98%) was achieved. The application of nitrifying biocath-
odes makes it possible to integrate the nitrogen and carbon removal
in MFC systems.
2.6. Miscellaneous species
Lovley et al. used Dessulfobulbus propionicus [53] and Geothrix
fermentans [54] for electricity generation. The Fe(III)-reducing
organism D. propionicus and G. fermentans conserved energy to
support growth by coupling the complete oxidation of acetate to
reduction of a graphite electrode. Geothrix cells were able to directly
reduce electrodes in the absence of an electron shuttle, but the
accumulation of a mediator over time further enhanced the rate of
electron transfer [54]. G. fermentans transferred more than 80% of
the electrons, available in their organic substrates, for the electric-
ity. Rabaey et al. [55] used microbial fuel cells for sulfide removal.
Microbial fuel cells were coupled to an anaerobic upflow ludge
blanket reactor, providing total removals of up to 98% and 46%
of the sulfide and acetate, respectively. The MFCs were capable
of simultaneously removing sulfate via sulfide. This demonstrated
that digester effluents can be polished by a MFC for both residual
carbon and sulfur compounds. The recovery of electrons from sul-
fides implied a recovery of energy otherwise lost in the methane
digester. They had isolated sulfide oxidizing organisms Paracoccus
denitrificans (with nitrate as the electron acceptor) and Paracoc-
cus pantotrophus. The Paracoccus species possess a membrane
bound complex that allows sulfide oxidation by the respiratory
chain.
Xing et al. [56] investigated the Rhodopseudomonas palustris DX-
1 for the electricity generation and found that it was capable of
high power production of the order of 2720 ± 60 mW/m2 . The vol-
ume size of the MFCs was 22 and 24 mL with 7 cm2 total exposed
surface area of the electrode. The internal resistance was approxi-
mately 8  and the external load was fixed at 1000 . Strain DX-1
utilized a wide variety of substrates (volatile acids, yeast extract,
and thiosulfate) for power production in different metabolic modes,
making it highly useful for studying the power generation in MFCs
and generating power from a range of simple and complex sources
of organic matter. Lewandowski et al. [57,58] operated a microbial
fuel cell in which glucose was oxidized by Klebsiella pneumoniae
[57,58] in the anodic compartment, and biomineralized manganese
oxides, deposited by Leptothrix discophora [58], were electrochem-
ically reduced in the cathodic compartment.
E. coli is also reported as the biocatalyst in the MFCs [59–62].
Zou et al. [59] constructed a MFC using polypyrrole coated car-
bon nanotubes (CNTs) composite as an anode material and E. coli
as the biocatalyst. In another report, Park and Zeikus [60] inves-
tigated MFC, where neutral red (NR) was utilized as an electron
mediator consuming glucose to study both its efficiency during
electricity generation and its role in altering anaerobic growth and
metabolism of E. coli and Actinobacillus succinogenes. Qiao et al. [61]
investigated the electrochemistry and electrocatalytic mechanism
of evolved E. coli cells in MFCs. Xi and Sun [62] reported the pre-
liminary study on E. coli based MFC and on-electrode taming of the
biocatalyst. The maximum power density reached 263.94 mW/m2
with the corresponding current density 1287.50 mA/m2 and the
internal resistance of MFC was 200 .
187
3. Conclusions and future prospects
MFC represents a potential method for bioelectricity genera-
tion and waste treatment although significant technical barriers
exist. MFC converts the chemical energy released in the oxidation
of organic wastes directly to electric energy. The detailed char-
acterization of the interfacial electron transfer rates, biocatalytic
rate-constants and cell resistances is essential upon the construc-
tion of the biofuel cells. Identification of the rate-limiting steps
allows then the development of strategies to improve and enhance
the cell output. The stepwise nano-engineering of electrode sur-
faces with relay-co-factor-biocatalyst units by organic synthesis
allows us to control the electron transfer cascades in the assemblies.
The production of electrical energy from biomass substrates
using biofuels could complement energy sources from chemical
fuel cells. Recently, the use of biocathodes in microbial fuel cells
solved various problems like requirement of a catalyst or artificial
electron mediators, which are very costly. These biocathodes can
produce useful products or remove unwanted compounds. There-
fore, the development of new methodologies could curb the length
of time needed for the realization of alternative sources of bioelec-
tricity. In addition, these developments will lead to clean and green
environment.
Acknowledgment
Authors specially acknowledge the reviewers of the manuscript
for enabling us to refine the manuscript.
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