Microbial Fuel Cell

Sediment-based MFCs are, due to their low complexity and low

power expectation, the type of MFCs that is closest to application.
From: Comprehensive Biotechnology (Third Edition), 2016

Related terms:

Bacterium, Biofilm, Proton, Mediator, Microorganism, Electron Transport, Electric

Potential

Environmental and Related Biotechnologies

M. Ruscalleda Beylier, ... R.-C. Wang, in Comprehensive Biotechnology (Third
Edition), 2011

6.22.4.1 Microbial Fuel Cells

Microbial fuel cells (MFCs) are a new bioelectrochemical process that aims to
produce electricity by using the electrons derived from biochemical reactions
catalyzed by bacteria. The energy generated by MFCs is expected to supply enough
energy to partially cover the energy demand in urban WWTPs.2
In MFCs, the electrons released by bacteria from the substrate oxidation in the
anode compartment (the negative terminal) are transferred to the cathode
compartment (the positive terminal) through a conductive material. In the cathode,
the electrons are combined with oxygen and the protons diffused through a proton
exchange membrane. MFCs require sustained electron release in the anode and
electron consumption in the cathode.17 The attainable metabolic energy gain for
bacteria is directly related to the difference between the anode potential and the
substrate redox potential. The optimal design for MFC is still under investigation,
and different materials for the electrodes as well as more selective membranes for
proton exchange are being currently developed to enhance their performance. It
seems that small cells connected in series offer higher potentials than bigger
reactor volumes. Nowadays, the main drawback for the full-scale application of
MFC is the cost of materials and the low buffering capacity of domestic wastewater.
For this reason, there is no industrial application of MFC to date.
However, the feasibility of domestic wastewater treatment by MFCs has been
successfully tested in laboratory experiments, obtaining COD removal up to 50%
and power densities about 420–460 mW m−2.2 Recently, C and N removal was
obtained in an MFC fed with synthetic wastewater containing and acetate.
Here, SND was performed in the cathode compartment separately from carbon
oxidation.27 This configuration optimized the C source and reduced the COD
requirements. Moreover, as denitrification uses the electrons obtained from the
separate oxidation of organic matter present in the wastewater, the MFC system
can operate very efficiently at low COD/N ratios. This reduces the requirement for
an external C-source supply. Thus, the results reported in the literature prove that
N removal with electricity production in MFCs is possible and bring the horizon of
self-sufficient WWTPs closer.

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Cyanobacteria-based microbial cell factories for

production of industrial products
Ragaa A. Hamouda, Noura El-Ahmady El-Naggar, in Microbial Cell Factories
Engineering for Production of Biomolecules, 2021

6 Microbial fuel cell

Microbial fuel cell (MFC) (Fig. 3) contains an anode and a cathode divided by a
cation exchange membrane. Microorganisms oxidize organic compounds in the
anode chamber, producing electrons and protons (reaction of anodes):
Fig. 3. Schematic diagram of two-chamber MFC (Anton et al., 2014).

The electrons are moved to the cathode across the exterior circuit, while the
protons are moved to the cathode across the membrane, oxidizing environment
current in the cathode chamber (supplied with oxygen). Electrons and protons are
finally spent in the cathode chamber, commonly reducing oxygen to water. Oxygen
is commonly operated as the electron acceptor for the cathode reaction in
microbial fuel cells (MFCs) (Anton et al., 2014):

or

Cyanobacteria have been used as sources of oxygen in the cathode chamber (Anton
et al., 2014; He and Angenent, 2006). The benefits of algal cathodes include
excluding the requirement of mechanical cathode air supply, which means lower
operating costs and lower overall CO2 emissions from the anodic bacterial
breathing and also harvesting algal biomass that is directly used as fuel feedstock
in the anode (Saba et al., 2017). Algal powders have been used previously as
feedstock for MFCs (Velasquez-Orta et al., 2009; Rashid et al., 2013) or pretreated
microalgae (Kondaveeti et al., 2014) and macroalgae (Gadhamshetty et al., 2013).
Certain studies display to use algae in MFCs rely on the hydrogen molecule
production, which is then oxidized at the anode for electron transfer to the MFC
circuit. Others have used algae as organic feedstocks to provide the electrons from
the oxidation of organic matter for MFCs to the electrochemically active bacteria
(Strik et al., 2008). Cyanobacterial isolates such as Anabaena and Nostoc have been
operated as biocatalysts in MFCs (Tanaka et al., 1985; Yagishita et al., 1997). The
maximum power density generated by photosynthetic MFCs (PMFC) using
Spirulina platensis as the biocatalyst reached 6.5 mW/m2 at high open circuit voltage
without externally added feedstocks (Fu et al.,2010). The maximum power density
generated by using Nostoc sp. ATCC 27893 in the anode of a PMFC reached
250 mA/m2 and 35 mW/m2 (Sekar et al., 2014).

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Constructed Wetlands for Urban Wastewater Treatment:

An Overview⁎
Diederik P.L. Rousseau, ... Rui Zhang, in Reference Module in Earth Systems and
Environmental Sciences, 2021

Crossover with other environmental technologies

Microbial fuel cell (MFC): MFCs are bioelectrochemical systems that generate
current by means of electrochemically active microorganisms as catalysts. In a
wastewater MFC, organic substrates present in the wastewater are hydrolyzed and
oxidized by bacteria. The electrons are transferred to an anode from where they
flow through a conductive material and a resistor to an electron acceptor, such as
oxygen, at the cathode (Logan et al., 2006; Rabaey et al., 2007). Such processes
require a redox gradient. A CW has aerobic and anoxic zones and is therefore
suitable for embedding MFCs (Ji et al., 2021). The technology is still in its infancy
and needs more research before upscaling. The power output is low and ranges
from a few to several thousands of milliWatts, however the combination of a
standard type of CW with an MFC yields better COD and nitrogen removal
compared with a standalone CW (Ebrahimi et al., 2021; Gupta et al., 2021). An
MFC can also be used as an alarm-like biosensor for detecting high COD influent
concentrations, as the voltage of an MFC is correlated with the COD concentration
(Corbella et al., 2019).
Green roofs: Green roofs are typically used to reduce stormwater flows by storing
rainwater, and in addition provide air filtration, cooling in urban areas and better
building insulation. However, they can also be constructed or converted into CWs
for treating domestic wastewater (Vo et al., 2019). The design generally consists of
a standard type HSSF or VDF CW. Surface flow CWs are not recommended as they
are prone to odor nuisances and mosquito growth. The depth of the CW is reduced
and lighter substrate material is required to reduce structural load on the roof. The
treated water can be reused for flushing toilets or irrigation as this poses less
health and safety issues than reuse for other purposes.
Green walls: Green walls or living walls have become popular to improve the
aesthetic appearance of buildings and to introduce patches of green in dense urban
environments. Similar to green roofs, they also provide some insulation in terms of
heat and noise, provide a habitat for insects and can improve air quality by
capturing fine dust. Whereas most green walls are maintained with tap water or
rainwater and chemical fertilizers, it has also been shown that such systems can
treat wastewater (Pradhan et al., 2019). Flipping a CW 90° obviously saves a great
deal of space.

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Electrochemical biosensors: Biomonitoring of food

adulterants, allergens, and pathogens
Pranveer Singh, in Electrochemical Biosensors, 2022

5.5 Biomonitoring of food-toxins

Food toxins are the metabolic by-products generated by fungi, algae, bacteria, and
plants. Mycotoxins or the fungal toxins are secondary metabolites of fungus of
which more than 400 types are already reported. Food infested with fungi shows
the presence of mycotoxins and other toxic metabolites leading to serious health
complications culminating in cancer. These toxins show a great diversity in their
chemical structure, physicochemical properties, and toxicological behavior (Marin
et al., 2013). Toxins can be carcinogenic, teratogenic, or allergenic. Detection of
these toxins is tricky as these are present in trace amount and occur in various
combinations either produced by single fungal species or multiple fungal species.
A folding mechanism-based electrochemical biosensor utilizing MB-tagged anti-
OTA aptamers was used for the determination of ochratoxin A (OTA) (Mishra et al.,
2015). In another work, aflatoxin M1 (AFM1) was identified in raw milk by
impedimetric biosensor using hexaethylene glycol-modified aptamer on screen-
printed-electrode (SPE) (Istamboulié et al., 2016) (Fig. 12).
Fig. 12. Schematics for bacteria and toxin detection using antibody-based
electrochemical biosensor (Mishra et al., 2018).
Copyright © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open-access
article distributed under the terms and conditions of the Creative Commons Attribution (CC BY)
license (http://creativecommons.org/licenses/by/4.0/).

Bioreceptor arrays contain electrodes functionalized with different bioreceptors

with specific binding targets. Optical and Piezoelectric sensors are employed for
the detection of toxins and chemicals. Fluorescent nanoparticles are developed to
detect surface-, inter- and intracellular toxins (Saucedo and Mulchandani, 2016;
Patra et al., 2017; Bahadır and Sezgintürk, 2017). Toxin separation and
quantification from food can be achieved by various sensing platforms in
conjunction with (SERS) (Moran et al., 2016; Liao and Lu, 2016; Stadler, 2016).
Okadaic acid (OA) is a major marine lipophilic biotoxin produced by Dinoflagellates
such as Dinophysis and Prorocentrum. Okadaic acid (OA) contaminated shellfish lead
to a physiological and pathological condition known as diarrhetic shellfish
poisoning that includes gastronomical disturbances (Callejas et al., 2015; Eangoor
et al., 2017). A voltammeter immunosensor having graphene-modified screen-
printed electrode (GSPE) detected OA using enzyme labeling (Eissa and Zourob,
2012).
An impedimetric aptasensor based AuNPs/rGO nanocomposite modified graphite
electrode was used for determining tetracycline in milk, meat, fish, and shrimp
(Mohammad-Razdari et al., 2020). Botulinum (BoNT) is a neurotoxin produced by
Clostridium botulinum. BoTN exists in seven serotypes (A-G), BoTN/A being the
most severe. It can lead to severe paralysis even in the lower dose. An rGO/Au
electrode-based biosensor was used to detect botulinum neurotoxin using
differential pulse voltammetry (DPV) (Chan et al., 2015).
5.5.1 Microbial fuel cells for food safety and security
The microbial fuel cells can be used for monitoring toxicity and biochemical oxygen
demand (BOD) in water, thus serving as an early warning system for food safety
and security. The sensing platform can be improvised for sensitivity, detection
range, and limit, integrating with detection algorithms, and empirical models to
differentiate toxicity from BOD on account of simultaneous occurrence (Jiang et al.,
2018). The microbial fuel generation cells relying on microbial interaction can
generate electricity through the oxidation of organic or inorganic wastes. Biofuel
production from the combination of plants, microbes, and algae can provide a
green and environment-friendly alternative to power and energy in the future
(Pinto and Ferranti, 2016).

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Environmental and Related Biotechnologies

C. Koch, ... F. Harnisch, in Comprehensive Biotechnology (Third Edition), 2016

6.43.3.2 Sediment MFCs

Sediment-based MFCs are, due to their low complexity and low power expectation,
the type of MFCs that is closest to application. Fig. 9 shows photographs of a
prototype, denominated as benthic unattended generator (BUG), based on this
simple, yet striking, concept. The anode is embedded in the (anoxic) sediment,
while the cathode is placed in the above sea water, where oxygen is available.
Generally, sediment MFCs yield only small current and power outputs. However,
these amounts of electric energy are typically sufficient to power small devices such
as radio sensors or meteorological buoys in remote areas and the deep ocean
(Tender et al., 2008; Thomas et al., 2013).
Fig. 9. Top: Meteorological data buoy used in demonstration on the pier of the
Naval Research Laboratory in Washington, DC, prior to deployment. Bottom: One
of the first-generation benthic MFC subunits on pier prior to deployment.
From Tender et al. (2008).

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Environmental Biotechnology
David P. Clark, Nanette J. Pazdernik, in Biotechnology (Second Edition), 2016

Microbial Fuel Cells

In microbial fuel cells, microbes such as bacteria catalyze electrochemical
oxidations or reductions at an anode or cathode, respectively, to produce an electric
current (Fig. 12.9). These fuel cells were originally inefficient and only served the
purpose of a battery in very remote areas. Advances in the understanding of the
microorganisms have increased the efficiency for the reactions.
FIGURE 12.9. Microbial Fuel Cell
(A) Schematic showing the cathodic and anodic chambers of a microbial fuel cell.
On the anode, microorganisms use organic matter such as wastewater or added
nutrients to create electrons, protons, and carbon dioxide. The electrons then flow
through the electric meter to the cathode. At the cathode, microorganisms can
convert the electrons to reduce oxygen to water under aerobic conditions, or
convert nitrate to nitrite or N2, or convert CO2 to acetate. These reactions can
create fuel precursors. (B) Actual microbial fuel cell showing the anode chamber
(left) and cathode chamber (right). These are separated by a membrane that allows
protons to freely pass from anode to cathode.
From Dolch et al. (2014) Characterization of microbial current production as a function of microbe-
electrode interaction. Bioresource Technol 157, 284–292.

Microbial fuel cells can harvest electricity from electrode-reducing organisms that
donate electrons to the anode. While the microorganism oxidizes organic
compounds or substrates into carbon dioxide, the electrons are transferred to the
anode. The mechanism of electron transfer can occur by three different pathways
(Fig. 12.10). First, electrons can be transferred to the anode through a soluble
mediator in the solution bathing the electrode. Second, electrons can be
transferred directly to the anode through proteins found on the outer membrane
of the bacteria. For example, microorganisms from the Geobacteraceae family
transfer electrons to electrodes using cytochromes on the outer membrane.
Shewanella oneidensis also uses cytochrome c to transfer electrons but requires an
anaerobic environment to convert lactate to acetate. In some instances, bacteria
form a thick film on the cathode, so it may be the pili or nanowires that transmit
the electrons to the anode. In contrast, electrode-oxidizing organisms use
electrons from the cathode to reduce substances in the cathode chamber. In
aerobic chambers, microorganisms can reduce oxygen to water. In anaerobic
environments, nitrate or sulfate can be reduced to nitrite, nitrogen, or sulfur ions.
Another potential reduction for these bacteria is the conversion of carbon dioxide
to methane or acetate. The process uses acetyl-CoA as an intermediate to build
even longer chain fatty acids and alcohols. For example, G. sulfurreducens reduces
fumarate to succinate with electrons obtained from the cathode. Interestingly, the
substrates that these organisms need for the redox reactions can be readily
obtained from wastewater or contaminated water, which would both provide
energy and clean up the environment.
FIGURE 12.10. Transfer of Electrons to the Anode in a Microbial Fuel Cell
Three different methods exist for bacteria to pass electrons from the oxidizing
reaction to the anode. There can be an extracellular mediator that absorbs the
electrons and passes them onto the anode (top). The bacteria can transfer electrons
through outer membrane proteins such as cytochrome c (middle). The electrons
can pass from the bacteria to the anode via nanowire structures such as pili
(bottom).

The use of microbial fuel cells is still not optimized, and the level of electric current
generated by such systems is low, but the potential for such systems is great. For
example, if a microbial fuel cell were to reduce carbon dioxide to make electricity,
not only would there be a renewable source of fuel, but the excess carbon dioxide
put into the atmosphere by burning fossil fuels could be used. The best
microorganism for producing an electric current is Sporomusa ovata, which is an
anaerobic, Gram-negative bacterium that converts hydrogen and carbon dioxide to
acetate by fermentation. In comparison to a standard hydrogen electrode, this fuel
cell produces −400 mV. However, the current generated is small. Current research
is now trying to identify what proteins are essential for the various reactions that
transfer electrons from the bacteria to the anode or take the electrons from the
cathode to reduce substrates. These types of studies should identify ways to
optimize the reactions to get the most energy from the bacteria. In addition,
researchers are still investigating the best materials for the cathode and anode, as
well as the solutions in which to grow the cells. Nevertheless, using microbial fuel
cells may help reduce environmental contaminants such as wastewater, reduce
atmospheric carbon dioxide by using it to rebuild fuels, and may potentially
provide a renewable energy source.

Microbial fuel cells create electricity through the use of microorganisms.

Organisms that transfer electrons to the anode are called electrode-reducing
organisms. They can pass electrons through a mediator molecule in the
solution, directly through proteins in their outer membrane, or through
nanowires or pili that coat the outer surface of the bacterium. Electrode-
oxidizing organisms take electrons from the cathode to reduce various
substances, such as carbon dioxide to acetate.

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Energy Storage Technologies and Their Role in
Renewable Integration and Significance of
Thermodynamic Analysis
Bahman Zohuri, in Encyclopedia of Energy Storage, 2022

Saliva powered battery

New battery is activated by your spit, saliva-Powered battery could be helpful in
extreme conditions.
Researchers have developed the next step in Microbial Fuel Cells (MFCs): a battery
activated by spit that can be used in extreme conditions where normal batteries do
not function.
For the last 5 years, Binghamton University Electrical and Computer Science
Assistant Professor Seokheun Choi has focused on developing micro-power
sources for the use in resource-limited regions to power Point-of-Care (POC)
diagnostic biosensors; he has created several paper-based bacteria-powered
batteries.
“On-demand micro-power generation is required especially for point-of-care
diagnostic applications in developing countries,” said Choi. “Typically, those
applications require only several tens of microwatt-level power for several minutes,
but commercial batteries or other energy harvesting technologies are too expensive
and over-qualified. Also, they pose environmental pollution issues.”
Choi, along with research assistant Maedeh Mohammadifar, created a high-
performance, paper-based, bacteria-powered battery by building microbial fuel
cells with inactive, freeze-dried exoelectrogenic cells which generates power within
minutes of adding saliva. The proposed battery generated reliable power from with
one drop of saliva, supplying on-board power that could be used by the next
generation of disposable, paper based POC diagnostic platforms.
“The proposed battery has competitive advantages over other conventional power
solutions because the biological fluid for on-demand battery activation is readily
available even in the most resource-constrained settings, and the freeze-drying
technology enables long-term storage of cells without degradation or
denaturation,” wrote the researchers. Choi is focused on improving the batter's
power density so that more applications can be powered.
“Now, our power density is about a few microwatts per centimeter square.
Although 16 microbial fuel cells connected in a series on a single sheet of paper
generated desired values of electrical current and voltage to power a light-emitting
diode (LED), further power improvement is required for other electronic
applications demanding hundreds of milliwatts of energy,” said Choi.

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Advances in Microbial Physiology

Derek R. Lovley, ... Kelly P. Nevin, in Advances in Microbial Physiology, 2011

11.3 Microbial Fuel Cells, Electrosynthesis, and Bioelectronics

There is significant interest in the development of large-scale microbial fuel cell
systems for wastewater treatment. Given the consistent enrichment of
Geobacteraceae on anodes of effectively operating microbial fuel cells, pre-
enrichment of anodes with Geobacter species may be an important step in scale-up
(Cusick et al., 2011).
There may be significant potential for increasing the current output of microbial
fuel cells via strain selection/design (Izallalen et al., 2008; Yi et al., 2009). The anode
of a microbial fuel cell is not a natural electron acceptor, and thus it is unlikely that
there has been significant selective pressure on Geobacter species to optimize
current production under the conditions found in microbial fuel cells (Lovley,
2006a). For example, increasing pilin expression of G. sulfurreducens, via strain
selection or genetic engineering, increased biofilm conductivity and current
production (Malvankar et al., 2011b). As more is learned about the mechanisms for
electron transfer to electrodes in Geobacter species, it may be possible to further
enhance power output.
Even without strain improvement there may be some short-term practical
applications for microbial fuel cells, such as powering electrical devices in remote
locations, such as at the bottom of the ocean (Tender et al., 2008). The fact that
Geobacter species are often the primary microorganisms colonizing electrodes
harvesting current from a diversity of environments suggests that they are likely to
play an important role in any applications of microbial fuel cells in which current is
harvested in open environments in which there will be competition for anode
colonization. Geobacter-based sensors may also be practical (Davila et al., 2010).
Novel system designs make it feasible to consider producing current with
Geobacter species, even in completely aerobic environments (Nevin et al., 2011b).
Electrodes deployed in subsurface environments are naturally colonized by
Geobacter species (Williams et al., 2010) and may function as sensors of subsurface
microbial activity (Tront et al., 2008; Williams et al., 2010).
Microbial electrosynthesis is a process in which electrons are provided to
microorganisms colonizing an electrode to support the reduction of carbon dioxide
to organic compounds that are excreted from the cells (Lovley, 2011b; Lovley and
Nevin, 2011; Nevin et al., 2010, 2011a). When powered with solar technology,
microbial electrosynthesis is an artificial form of photosynthesis in which sunlight
drives the conversion of carbon dioxide and water to organic compounds and
oxygen. Proof-of-concept studies have demonstrated acetate production with
acetogenic microorganisms as the catalysts (Nevin et al., 2010, 2011a).
Genome annotation led to the surprising discovery of enzymes for carbon dioxide
fixation in some Geobacteraceae (Aklujkar et al., 2010). Within the G. metallireducens
genome, a pair of genes is predicted to encode an ATP-dependent citrate lyase,
which would allow the reverse TCA cycle to produce acetyl-CoA. Further, genes for
all of the identified enzymes of the dicarboxylate/4-hydroxybutyrate cycle of carbon
dioxide fixation are predicted in the G. metallireducens genome. G. metallireducens is
also capable of electrosynthesis, and investigations with genetically modified
strains of other Geobacter species are ongoing because of the ability of Geobacter
species to interact so effectively with electrodes. G. sulfurreducens can also use
electrons derived from an electrode to reduce protons to hydrogen (Geelhoed and
Stams, 2011), potentially providing a renewable catalyst that is much less expensive
than the metal catalysts typically employed for hydrogen production.
One of the most exciting practical applications for Geobacter species could be
bioelectronics. Electronically functional biomaterials are very attractive because
they can be synthesized from relatively inexpensive feedstocks and do not contain
toxic components (Hauser and Zhang, 2010). Further, conductive materials
comprising living bacteria are self-renewing because bacteria can self-repair and
replicate. Initial studies have already demonstrated the possibility of tuning the
electronic properties of Geobacter biofilms via simple genetic engineering and
more sophisticated modifications are feasible. Further elucidation of the
mechanisms for electron transport along pili and ability of cytochromes to function
as capacitors could aid in the biomimetic design of new materials. Therefore, it is
expected that microbially based electronically functional materials will have
significant potential for next-generation biotechnological applications.

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Biofuel cells as sustainable power sources for

implantable systems
S. Kerzenmacher, in Implantable Sensor Systems for Medical Applications, 2013
6.6 Sources of further information
An excellent overview on various scientific and technological aspects of enzymatic
and microbial fuel cells is provided in the book ‘Bioelectrochemical Systems: from
extracellular electron transfer to biotechnological application’ edited by Korneel
Rabaey et al. (2010). A detailed treatise on the history and technology of
implantable abiotic glucose fuel cells is available from Kerzenmacher et al. (2008b).
Similarly, a number of review articles on enzymatic and microbial fuel cells are
available (Bullen et al, 2006; Davis and Higson, 2007; Cooney et al., 2008;
Moehlenbrock and Minteer, 2008). The theoretical background of electrochemical
energy conversion and methods for the study of electrochemical systems is
described in detail in the book ‘Electrochemistry’ by Hamann et al. (2007). For
further reading on other applications of fuel cells, the book ‘Fuel Cell Systems
Explained’ by Larminie and Dicks (2000) is recommended.

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Emerging Trends of Microorganism in the Production

of Alternative Energy
Golla Ramanjaneyulu, Bontha Rajasekhar Reddy, in Recent Developments in
Applied Microbiology and Biochemistry, 2019

21.3.8 Microbial Fuel Cells

Transformation of chemical energy to electric energy is known from eighteenth
century of Volta, the inventor of voltaic pile and who was the contemporary of Luigi
Galvani who initially observed animal electricity. In the presence of biological
catalysts like enzymes (enzymatic fuel cells) and microorganisms (microbial fuel
cells, MFCs), the chemical energy accessible in biomass surrounding us can be
harnessed. Many researchers have started focusing on the ability of
microorganisms to produce electric energy in biological systems (Potter, 1910). In
1911, Potter observed that a maximum voltage of 0.3–0.5 V could be generated
with glucose as a substrate and Pt (platinum) as electrode by the S. cerevisiae.
Presently, for almost a century, research is continuously progressing on MFCs by
the oxidation of organic matter to produce electric energy providing a great scope
toward alternate energy (Pant et al., 2012). Bioelectrochemical cells (BEC) have
gained significant interest in the production of bioenergy from natural biomass
and wastewaters. In particular, microbial fuel cells (MFCs) and microbial electrolysis
cells (MECs) have been extensively exploited for bioelectricity and biohydrogen
generation (Logan et al., 2015). From a biological perspective, both kinds of fuel
cells work on a similar principle; consequently, common microorganisms can be
deployed in these fuel cells in bioenergy production. The distinctive character of
these microorganisms (referred as exoelectrogens or electricigens) in BEC is the
display of particular molecular machinery that helps exchange the electrons from
microbial outer membrane to the conductive surfaces (Kumar and Kumar, 2017).
Hence, the electrons can be utilized to produce electricity and hydrogen. MFCs are
one of the widely studied technologies that have potential for waste valorization
into energy in the form of bioelectricity production (Koók et al., 2016). In spite of
critical progression occurring in this field with respect to microbiology, materials
science, chemistry, electrochemistry, etc., process economization and process
sustainability were observed to be the most essential elements to move the field to
the next level (Mohanakrishna et al., 2012). Microorganisms present in MFCs as
catalysts to drive to the anodic and cathodic reaction to generate electricity. By
exoelectrogenic microorganisms, biodegradable substances containing chemical
energy can be converted into electricity. At the anode compartment, electrons and
protons are produced by the oxidation of organic compounds by certain microbes.
The electrons reached to the cathode compartment through the external circuit
where it reduces the electron acceptor present, and in the meantime, protons
produced at the anode are exchanged to the cathode through a membrane
separator or through the electrolyte in a membraneless cell.
The anodic reaction that is based on microbial oxidation is similar for all types of
MFC applications. However, different cathodic reactions can be employed in MFCs
to generate electric energy if the overall reaction is thermodynamically favored.
Because of unlimited availability and positive redox potential, oxygen is mostly
widely considered as the favorable electron acceptor for practical applications. The
attainability of utilizing other electron acceptors with a high redox potential, for
example, nitrate, sulfate, and some other contaminants in the environment with
high redox potential, which are electrochemically or naturally reducible in the
cathode chamber, can also be considered (Berchmans, 2018). Additionally, to
increase the voltage of the cell, permanganate, dichromate, peroxide, and
ferricyanide are being used as a part of MFCs in light of their high redox potential
(Yang et al., 2011). Microorganisms that can reoxidize reduced metal oxides and
Fe2 + species like Acidithiobacillus ferrooxidans and Thiobacillus ferrooxidans can also
be utilized as cathodic biocatalysts (Kumar et al., 2015).
In MFCs, the anode and cathode are isolated by an ion-exchange membrane, and
solutions comprising biomass and microorganisms are used as fuel (Logan and
Regan, 2006; Lal, 2013):
Anode : C6H12O6 + 6H2O → 6CO2 + 24H+ + 24e−
Cathode : 6O2 + 24H+ + 24e− → 12H2O
C6H12O6 + 6O2 → 6CO2 + 6H2O + Electric Energy
However, the outputs of energy from MFCs and MECs are inadequate for
industrial-level applications and, therefore, not feasible for commercialization.
Hypothetically, an MFC can create a maximum voltage of 1.2 V, and the optimum
hydrogen generation yields in MEC would be 3.4 mol H2/mol acetate (Logan et al.,
2015). Initially, the research on MFC was focused on wastewater treatment;
however, over a period of time, the field of MFCs has developed into a much more
diverse field of research called bioelectrochemical systems (BES) because of the
advent of several related technologies such as microbial electrolysis, microbial
desalination, microbial electrosynthesis, and photomicrobial cells. Latest studies by
the use of distinctive electrode materials and also MFC reactor designs in the scope
of 200 mL to a couple of hundred liters were assessed toward the advancement of
pilot-scale MFC systems (Janicek et al., 2014). Transforming this technology from
lab scale to industrial scale will bring it a step toward the realization of commercial
application of bioelectricity generation. Among the few materials used for the
electrodes, carbon-based materials, like graphite and carbon with metal-based
impregnations or coatings, were observed to be economical and proficient for
bioelectricity production (Xie et al., 2010; Mohanakrishna et al., 2012).
If MFCs are benchmarked against anaerobic digestion, two distinct differences are
to be observed. MFCs cannot deal with suspended and particulate organic material,
though anaerobic assimilation is capable of dealing with them. Using MFCs other
than electricity, many other value-added products can be obtained that is otherwise
not possible with anaerobic digestion. However, the rates of energy conversions are
lower in MFC technology. Thus, all the technological challenges need to be clearly
understood to make the MFC technology more viable. Proper power management
systems should be evolved to maximize the power output derived from MFCs and
to integrate with MFC. Synthetic biology may help in developing robust
exoelectrogens with perfect electron-exchange properties. MFC, as energy-saving
technology, may well wean for us far from the dwindling oil assets. Before that,
there are many technical challenges that must be considered for sustainable and
renewable energy generation. Regardless, the technology may open the way to new
method for renewable and sustainable energy products. In the near future, MFCs
might be developed to such a phase, to the point that they can give a sensible and
usable power yield per unit the MFC volume. In such scenario, a larger battery size
could be ignored, provided the maintenance is simple and has a green and safe
label. This eco-friendly fuel cell will then lead to several groundbreaking
applications. As the amount of low-power devices implanted in the human body
increases, the long term, stable power source used may well be the MFC (Table
21.5).

Table 21.5. Different Applications of Metabolomic-Based Analyses to Biofuel

Area Major Findings Metabolomics Bioproduct(s) Organism
Methods

Pathway Metabolite LC-MS Isoprene- Escherichia c

engineering quantification detected derived
accumulation of alcohols, 3-
isopentenyl methyl-2-
pyrophosphate, butenol, 3-
indicating that NudB methyl-3-
was a bottleneck butenol, and
enzyme in engineered 3-methyl-1-
heterologous MVA butanol
pathway. Increased
NudB expression
resulted in a 60%
increase in methyl
butanol production

Time-dependent GC and GC- 1,4- Escherichia c

metabolite profiling MS Butanediol
was used to verify
enzymatic activity of
engineered xylose to
butanediol pathway
before regulation of the
pathway was optimized
Area Major Findings Metabolomics Bioproduct(s) Organism
Methods

Integrated omics was LC-MS, 1,4- Escherichia c

used to design and dynamic Butanediol
optimize a succinyl-CoA isotopic
to butanediol pathway. labeling
Dynamic labeling
showed that aldehyde
dehydrogenase was a
rate-limiting step,
guiding targeted
enzyme engineering
that resulted in a 20%
increase in titer.
Competing TCA cycle
reactions were
identified using
targeted
transcriptomics,
directed by isotopic
labeling

The mevalonate (MVA) LC-MS, Isoprene Escherichia c

and methylerythritol steady-state
phosphate (MEP) isotopic
pathways were found to labeling, 13C
be synergistic in MFA
isoprene production.
Dual overexpression of
MAV and MEP
pathways resulted in
enhanced productivity
beyond the expected
additive effect of
individual
overexpression, with a
final titer of 24 g/L
Area Major Findings Metabolomics Bioproduct(s) Organism
Methods

Medium-chain GC-MS, (3E,5E,7E)- Nigrograna

hydrocarbon steady-state Nona- mackinonnii
nonatetraene was isotopic 1,3,5,7-
found to be synthesized labeling tetraene
via a polyketide-
synthesis-related
pathway whereby head-
to-tail condensation of
acetate is followed by
decarboxylation.
Candidate polyketide
synthases were
identified

Redox Yeast responded to GC-MS and Any Saccharomyc

balancing increased NADPH IE-MS/MS, cerevisiae
demand by increasing steady-state
acetate production and isotopic
rerouting flux toward labeling, 13C
the PPP. These MFA
responses were
metabolically or
transcriptionally
controlled depending
on these varieties of
NADPH demand

Constitutive expression GC-MS Ethanol Fusarium

of oxysporum
phosphoglucomutase
and transaldolase
increased ethanol yield.
Metabolomic analysis
identified increased
NADPH availability
leading to reduced
acetate production as a
major source of
improvement
Area Major Findings Metabolomics Bioproduct(s) Organism
Methods

Increased carbon and GC-MS, Fatty acids Escherichia c

redox demands of fatty steady-state
acid over production isotopic
resulted in increased labeling, 13C
flux through the MFA
oxidative PPP and
increased conversion of
NADH to NADPH by
transhydrogenases. ED
glycolysis increased
only lightly despite its
ability to generate both
NADPH and acetyl-coA
for fatty acid synthesis,
making ED glycolysis a
potential target for
metabolic improvement

Increased carbon and GC-MS, Mevalonate Escherichia c

redox demands of steady-state
mevalonate over isotopic
production were met by labeling, 13C
conversion of NADH to MFA
NADPH via
transhydrogenase. This
study did not see
increases in oxidative
PPP flux, suggesting
that the moderate
NADPH demands for
mevalonate production
could be met by
transhydrogenase
reactions alone
Area Major Findings Metabolomics Bioproduct(s) Organism
Methods

A correlation was GC-MS, Any Pichia pastor

observed between steady-state
increased PPP flux and isotopic
increased expression of labeling, 13C
recombinant protein. MFA
Combined
overexpression of
glucose-6-phosphate
dehydrogenase and 6-
phosphogluconolactone
resulted in the highest
PPP flux and the
highest expression
levels of recombinant
protein

Central Flux modeling of GC-MS, Any Thermus

metabolism central carbon steady-state thermophilus
metabolism verified the isotopic
absence of ED labeling, 13C
glycolysis and oxidative MFA
PPP and showed high
TCA cycle flux

Flux modeling of GC-MS, Butanol, Clostridium

central carbon parallel ethanol, and acetobutylicu
metabolism revealed steady-state acetone
noncanonical TCA cycle isotopic
reactions, generation of labeling, 13C
C1 from pyruvate, and MFA
isoleucine production
via citramalate synthase

Flux modeling of GC-MS, Any Geobacillus s

central carbon parallel
metabolism showed steady-state
that the TCA cycle and isotopic
oxidative PPP are labeling, 13C
responsible for NADPH MFA
production during
growth on xylose
Area Major Findings Metabolomics Bioproduct(s) Organism
Methods

13C fingerprinting GC-MS, Any Rhodococcus

based on labeling parallel opacus PD63
patterns of only a few steady-state
amino acids was used isotopic
to assess the metabolic labeling, 13C
activity of EMP and ED fingerprinting
glycolysis,
gluconeogenesis,
glyoxylate shunt,
anaplerotic pathways,
and amino acid
synthesis in a
nonmodel organism

Xylose Expression of GC-MS, Ethanol Saccharomyc

utilization heterologous xylose steady- state cerevisiae (XR
reductase (XR), xylitol isotopic XDH xylose
dehydrogenase (XDH), labeling, 13C assimilation)
and xylulose kinase MFA
enzymes led to
increased flux through
the oxidative PPP and
TCA cycle to meet
increased NADPH and
energy demands,
limiting ethanol
production

Yeast strain with xylose GC-MS, Ethanol Saccharomyc

isomerase (XI)-based steady- state cerevisiae (XI
xylose assimilation did isotopic xylose
not exhibit high flux labeling, 13C assimilation)
through oxidative PPP MFA
suggesting that XI
ameliorates the redox
imbalances seen in XR-
HDH strains. However,
bottlenecks in lower
glycolysis limit ethanol
production
Area Major Findings Metabolomics Bioproduct(s) Organism
Methods

During sugar GC-MS, Butanol, Clostridium

coutilization, hexoses parallel ethanol, and acetobutylicu
were assimilated via steady-state acetone BOH3
glycolysis, while isotopic
pentoses were labeling, 13C
incorporated in to the MFA
PPP. The
phosphoketolase
pathway plays an
important role in
pentose metabolism
and could be targeted
for strain improvement

In xylose-utilizing strain GC-MS, Any Thermus

developed via directed parallel thermophilus
evolution, NADPH steady-state developed via
production was isotopic adaptive
identified as a limiting labeling, 13C evolution of s
factor during growth on MFA HB8
xylose, suggesting that
expression of
heterologous oxidative
PPP enzymes may
improve strain
performance

Stress Acetic acid was found to GC-MS and Ethanol Saccharomyc

tolerance inhibit xylose CE-MS cerevisiae (XR
fermentation due to an XDH xylose
accumulation of assimilation)
intermediates of the
nonoxidative PPP.
Overexpression of
transaldolase relieved
this bottleneck and
improved ethanol yields
Area Major Findings Metabolomics Bioproduct(s) Organism
Methods

Proline and myoinositol GC-MS, Any Saccharomyc

were identified as key regression cerevisiae
metabolites in tolerance modeling
to furfural, acetic acid, (PLS-DA)
and phenol.
Accumulation or
supplementation of
these metabolites
mitigates growth
inhibition

Phenolic amides inhibit LC-MS, Any Escherichia c

nucleotide biosynthesis dynamic
via competitive isotopic
inhibition of glutamine labeling
amidotransferase.
External
supplementation with
nucleosides mitigates
growth inhibition

Threonine was GC-MS, 1-Butanol Saccharomyc

identified as a key regression cerevisiae
metabolite contributing modeling
to butanol tolerance (OPLS)
based on
metabolomics-based
regression modeling.
Gene deletions aimed
at increasing threonine
accumulation resulted
in improved butanol
tolerance, providing a
proof of concept for
semirational
engineering based on
metabolomics data
 Area Major Findings Metabolomics Bioproduct(s) Organism
Methods

Directed evolution for GC-MS and 1-Butanol Methylobacte

improved butanol LC-MS extorquens A
tolerance resulted in
increased abundance of
disaccharides and
saturated fatty acids
and decreased levels of
carotenoids and
carotenoid precursors,
suggesting that
membrane fluidity and
osmotic control are
important factors in
butanol tolerance

Data from Martien, J.I., Amador-Noguez, D., 2017. Recent applications of

metabolomics to advance microbial biofuel production. Curr. Opin.
Biotechnol. 43, 118–126.

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URL: https://www.sciencedirect.com/science/article/pii/B9780128163283000210

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