JUNE 20, 2022

ROLE OF MICROBES IN CLEANING THE

ENVIRONMENT (MICROBIAL FUEL CELLS)
(ENVIRONMENTAL BIOTECHNOLOGY ASSIGNMENT)

SYED MUHAMMAD ZAKI HAIDER (BSBT025F18)

SUBMITTED TO: DR. BEENISH MAQSOOD
School of Biochemistry and Biotechnology, University of the Punjab
 ROLE OF MICROBES IN CLEANING THE ENVIRONMENT
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S
ince the inception of planet earth and life residing in it, energy has always been an
essential, a necessity for life and the whole ecosystem to thrive. The primary source of
energy for all living organisms (including us, the human beings) is sun, a rudimental
high school level concept we are all aware of. Talking about human beings specifically, over the
course of time, we evolved (and are still evolving). Our first and foremost society was the
hunter-gatherer one, totally dependent and connected with nature. This relatively much simpler
lifestyle continued to evolve into what we now know as the post-industrial era. Increased
standards of living, immensely increasing world population (currently standing beyond 7.8
billion and expected to go up to 9.9 billion by 2050), monolithic buildings and skyscrapers,
coalition of heavy duty industries and cutting edge technological developments, etc. All of this
by virtue of accolades in the fields of science and technology, over the course of decades and
centuries. We, humans, continued to experience different problems and work on their solutions.
We faced the problem of travelling longer distances in time effective way, we made vehicles. We
faced the problem of mass production of different machines, food, things to support our
relatively high standards of living, we set up industries for that. It is quite obvious that the
increment in our population and the living requirements (of such a gigantic population) also
comes with gigantically increasing energy requirements. Energy to run our factories, vehicles,
stoves, energy to brighten up our houses (or even build houses), and the list goes on. Now all this
industrialization, urbanization and consequently, the prosperity of nation somehow harmed our
environment.

We all know that energy is mainly classified as non-renewable or renewable. As the name says it
all, non-renewable energy sources cannot be replenished once they get completely used up (or
they do so but at a very slow, gradual rate). These include the fossil fuels (coal, natural gas,
crude oil). It takes millions of years in the formation of fossil fuels by the remains of dead cells
of plants and animals, under the earth’s crust. Fossil fuels are advantageous in ways that they are
traditional sources of energy that are easy to store, high in energy and very profitable, but the
cons outweigh the pros. Formation of fossil fuels, first of all, is very time consuming and at the
rate we are using them, most probably they will be wiped out altogether. From the procedures of
their extraction to their combustion, there is also an adverse increase in pollution which

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represents a major concern as far as environmental health and biodiversity are concerned.
Polluting air, water, land (directly or indirectly), the most concerned pollutants are the green-
house gases. And then come tiptoeing in the scene global warming, rising sea levels, extreme
climate changes, loss of habitats and consequently, biodiversity. In essence, environment and
energy are of great interdependence[1].

Therefore, there is an urgent requirement to develop a method which can generate renewable and
safe energy without the emission of net CO 2. To provide clean and freshwater to the modern
world, which is responsible to cover the basic needs of life is also a big task of 21st century.
Water is a major part of substance for all living organisms on earth. Water is also called the
universal solvent because it has ability to dissolve many substances. Therefore, polluted water
has unwanted minerals and chemicals that have adverse effects on human health and cannot be
used for any direct purpose. Nowadays, natural water sources have become contaminated due to
different factors like high residential ratio, commercial factors, industrial demand, improper
irrigation system, agricultural wastes, global warming, and medicinal waste, etc. The effect of
these factors on natural sources of water has resulted in a shortage of freshwater to maintain a
healthy environment for living organisms.

Aiming at environment conservation and remediation, multitudinous endeavors have been

developed and probed into. Talking about aquatic environment, municipal wastewater and
industrial effluent are considered to be decontaminated before discharging into receiving water
bodies, such as activated sludge treatment, membrane technology, anaerobic digestion, ion
exchange and electrochemical oxidation, etc. Whereas, there are some associated shortcomings
cannot be neglected for instance, energy demand, that more than 3% of global electricity
generation are devoted to wastewater treatment. Recently, the World Health Organization
(WHO) reported that there were more than 1.7 million deaths and near four billion suffering
from different diseases due to water pollution. Therefore, it is indispensable to treat wastewater
to overcome the environmental pollution problem and save the ecosystem.

There were many traditional methods reported for bioremediation of different toxic organic
compounds and metals that have adverse effects on living organisms. These conventional
methods include ozonation, electrolytic reduction, in situ and ex-situ treatment, thermal
treatment, chemical precipitation, in-situ confinement [2]. The above-mentioned methods are

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quite efficient, but they have several drawbacks like no proper electron acceptor or donor’s
mechanism and are quite expensive, such that they are not easy to maintain at a commercial
level. Moreover, all these conventional methods have prolonged process of degrading of organic
pollutants by catalyst (microbes).

We need smarter alternatives. We need renewable energy sources. And we need technologies

that are friendlier to the environment. One of these smart alternatives could come from the
oldest, smallest, and most adaptable creatures on the planet: microorganisms [3].

Microbial fuel cells:

The concept of producing electricity using microbes was first developed in 1917, using
microorganisms Escherichia coli [4]. MFC is an innovative, eco-friendly, and cheap method to
generate electricity along with water treatment. MFCs is quite a promising and developing
research field for the scientific community to transform chemical/organic energy into electrical
energy through using microbes. This technology has significant potential to make renewable
energy by utilizing organic waste.

Like all living organisms, microbes need energy and they do so by cellular respiration. We know
that during photosynthesis, autotrophic organisms like plants turn CO 2 and water into the sugars
they use for energy. Cellular respiration is essentially this process in reverse. It breaks down
sugars into water, CO2 and (most importantly) adenosine triphosphate (ATP). ATP is the energy
molecule that all cells need to function, thus called energy currency of cell. MFC basically is a
way of trapping this energy released by cellular respiration and breakdown of organic
compounds.

MFC is a bio-electrochemical system that utilizes the cellular respiring power of microorganisms
for the conversion of chemical energy present in the form of organic matter in the wastewater to
electrical energy [5]. MFCs come with a 2 way advantage, first one is the production of cheap
electricity using renewable sources (organic waste utilized by microbes in the process), second is
the treatment of waste water which is good for living things and the environment.

To understand more about MFC, we will start from the basics first. We will first look at how an
everyday battery works.

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Working of a battery:
A battery generates electricity using chemical energy. In a
battery, one of the electrodes (the anode) undergoes an
oxidation reaction. This is a chemical process that gives off
electrons. The electrons travel through a substance called
electrolyte (a paste in a dry cell or a liquid in a wet cell) to
the other electrode (the cathode). Here, the electrons undergo
a reduction reaction. This is a chemical process in which
electrons are absorbed. As the electrons move from anode to Figure 1 working of a battery

cathode, they create an electric current. This can do work, such as lighting up a bulb.

Traditional fuel cells:

A fuel cell is a device that generates
electricity through an electrochemical
reaction where the hydrogen gas and oxygen
gas combine to produce water and electrons
given out in the process move through the
electrode to generate electricity [6]. A fuel is
different from a battery in the aspect that it
generates energy from a fuel rather than
using stored energy [7]. The fact that it
involves mainly hydrogen as fuel and does Figure 2 working of a fuel cell
not undergo combustion like fossil fuels is something in support of fuel cells. Traditional fuel
cells have been successfully applied in a large scale. However, due to utilization of noble metals
as catalysts like Nickel and Platinum to generate electrons from fuel oxidation, traditional fuel
cells are facing many challenges such as high cost, lack of metal catalyst, dependence on
petroleum products (i.e. hydrogen, ethanol, methanol) and emission of CO 2 in formation of
hydrogen gas from methane, adding to green-house effect. Whereas an MFC does not face these
problems as we will see in its structure and working in detail.

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Structure of an MFC:
Generally, MFCs has two chambers consisting of a cathode and anode, respectively. The anode
chamber is enclosed into wastewater solutions (heavy metal or organic solutions) and other
(cathode) in surface water. The aerobic cathode chamber is separated by an ion exchange
membrane (e.g. anion, cation and proton exchange membranes) from the anaerobic anode
chamber [5]. In membrane-less MFC, a glass wool or beads (porous proton exchange system) is
used instead of IEM. In the anodic chamber, an oxygen deficient environment is required for
microbes to degrade organic waste and produce electrical energy.

In general, there are two types of MFCs: reactor MFCs (A) and sediment MFCs (B) [8]:

Figure 3 2 main types of MFC, reactor MFC and sediment MFC

Reactor MFCs are engineered systems that are designed and operated for energy-efficient
water/wastewater treatment or (after appropriate modification) production of specific compounds
such as hydrogen gas (microbial electrolysis cells) and value-added compounds (microbial
electrosynthesis cells). Wastewater treatment is a well-known function and objective for the
development of reactor MFCs, and tremendous efforts have been made to understand and
improve reactor MFCs from the aspects of microbiology, electrochemistry, materials, and

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configuration. The technology has been advanced with long-term operation of MFCs installed in
wastewater treatment plants and scaled up to several hundred liters.

Sediment MFCs (SMFCs), also referred to as benthic MFCs in some cases, are deployed in a
natural system or where there is less engineering management (e.g., a constructed wetland).
Unlike reactor MFCs that have a clear boundary between the anode and the cathode by using
membranes or separators, SMFCs rely on a naturally occurring oxygen gradient to separate the
anode and the cathode.6 To achieve that, the anode electrode is embedded in sediment where
dissolved oxygen (DO) is depleted, while the cathode electrode is installed in the water phase
with relatively higher DO.

Functioning of an MFC:
In the anode compartment, organic matter, acts as electrons donor, is oxidized into CO2 and
water by a biofilm of microbes present on it. Organic matter oxidation generates electron (e−),
C02 and protons (H+). Electrons will flow to cathode compartment through electrical circuit,
whereas protons flow through the membrane to the cathode (or beads in case of absence of
membrane). In cathode compartments, electron acceptors are reduced, thus forming water (H 2O).
Electrical power is coupled by placing a load between two electrode compartments (such as a
light bulb for instance). In MFC, electrotrophs microbes (also called exoelectrogens) convert
toxic compounds into less toxic components. Thus, electricity is being generated from waste

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water and it is also cleansed as well. A schematic diagram below show the overall fundamental
process taking place in an MFC:

Microbes used in MFC:

Microbes that transfer the electrons extracellularly are called exoelectrogens. For efficient
electon transfer, these exoelectrogens are present on the surface of the anaerobic anode in form
of a biofilm (thus sometimes also called anodophilic consortium of microbes). In systems in
sediments from 50 to 90% of the microorganisms at the anode are -Proteobacteria [9]. Some
fungi and algae are also known to have electrogenic properties. To a lesser extent Cytophagales
(to 33%), Firmicutes (11.6%), J-Proteobacteria (9-10%) [9]. Examples of gram-positive bacteria
in MFC include Clostridium butyricum, Bacillus cereus, Corynebacterium sp, and Thermincola
ferriacetica. [9]. While Escherichia coli, Pseudomonas fluorescens, Proteus vulgaris,
Pseudomonas methanica, Gluconobacter oxydans, Klebsiella oxytoca, Enterobacter sp and
Shewanella oneidensis fall under gram negative group of electrotrophic bacteria [9]. Based on
the conditions that are created in the anode chamber (the absence of oxygen), either anaerobic or
facultative anaerobic microorganisms can work there. Anaerobes include Geobacter
sulfurreducens, Geobacter metallireducens, Rhodoferax ferrireducens, Clostridium butyricum,
Enterobacter sp. Facultative anaerobes include Escherichia coli, Saccharomyces sp.,
Saccharomyces cerevisiae, Sarcina lutea, Proteus vulgaris, Desulfuromonas acetoxidans,
Shewanella oneidensis, Shewanella affinis, Bacillus cereus.

Electron transfer by exoelectrogens in MFC:

For the generation of electricity, exoelectrogens can transfer electrons from electrodes through
mechanisms such as short-range electron transfer through redox-active proteins, soluble electron
shuttling molecules, and long-range electron transport by conductive pili, and direct electron
transfer. The direct electron transfer means a direct contact of microbes and electrode surface.
There is no mediator or source to transfer the electrons to the anode surface in this case. The
better way is long-range electron transfer via conductive pili.

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 Direct electron transfer (DET) is a phenomenon in which physical contact of microbes

and electrode surface takes place through nanowires and redox-active proteins (Fig. 4).
Electrogenicmicrobes near electrodes surface develop nanowires (flagella and pili)/
biofilm, through which transfer of electron occurs directly from the outer cell membrane
without the requirement of any mediator. Redox-active proteins such as cytochromes at
the outer side of the cell membrane are also used for DET. Nanowires forming bacteria
such as Shewanella and Geobacter are preferred in MFC for their efficient electric power
production capacity. The role of the final electron acceptor can be played by the MFC
anode.
 Mediator electronic transfer (MET) is also called indirect electron transfer where
microorganisms are electrochemically inactive and cannot directly transfer electrons to
the electrode. In this case, soluble substances known as redox mediators can be used that
facilitate electronic transport. To ensure effective transfer of electrons from the
microorganism to the anode, the mediators must satisfy a number of special
requirements: to ensure fast and reversible transfer of electrons from the biocatalyst to the
electrode; have an oxidation-reduction potential close to the potential of the biocatalyst;
should be chemically stable. Several natural or synthetic mediators such as phena zine,
pyocyanin, anthracenedione, thionine, neutral red, humic acid, riboflavin, and methylene
blue have been used in MFC. Electrochemically active microorganisms in MFC are able
to produce their own mediator connections under certain conditions, which can be
involved in extracellular electron transfer processes. This can happen in two ways: by
producing secondary and primary metabolites. Secondary metabolites (endogenous
mediators) are redox active substances and serve as reversible final electron acceptors

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that transport electrons from a bacterial cell to a solid oxidant (anode of MFC) or to
aerobic layers of biofilm where they are oxidized and can again participate in redox
processes. Primary metabolites, for example sulphide or hydrogen, also serve as redox
mediators.

Types of MFCs:
Based on design and configuration, MFCs have been classified into single chamber, double
chamber, up-flow or tubular and stacked:

 The double chamber is one of the simplest designs of MFC. The DMFC consists of
anodic and cathodic chambers which are separated via a salt bridge. The anodic chamber
comprises of microbes and substrate (glucose, acetate, wastewater, etc.) and the anode
electrode. In the cathodic chamber, the cathode and catholyte such as freshwater, and air
are present for the consumption of e-.
 Single chamber MFC (SMFC) consists of anode and cathode within a single-chamber
without any aeration system or O2 supply. At the one end, a cathode is exposed to the
atmosphere and only anode with a substrate and microbial culture is present inside the
chamber. This simple and economical design of MFC is preferred over the double
chamber for better performance as smaller chamber increases volumetric power density
and reduces the extra operational cost required for aeration. The process scale-up for
operation in continuous or batch mode becomes easier in SMFC.
 Up-flow MFCs are cylindrical in shape and used for a continuous mode of operation. The
anode is attached to the lower end, whereas cathode is placed at the top side of the
cylinder, which is apportioned via glass wool or beads. A substrate such as wastewater
flows from the bottom, and effluent flows out from top through a series of anode and
cathode. In this set-up, the aeration system and the pump are required while the
operational cost is optimized due to the absence of a proton exchange membrane. They
are preferred for wastewater treatment due to ease of scale-up.
 In the stacked system, several MFCs are connected in a series or parallel, which results in
the multiplied power output, but some loss of voltage also occurred in some cases.
Various other operational factors can increase or reduce the overall performance of the

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set-up. Parallel stacked MFCs show better current generation and better COD removal
efficiency then series arrangement.

Waste water treatment using MFC:

MFC efficiently removes a wide variety of pollutants such as organic matter, nutrients,
pharmaceuticals, fats/oil, heavy metals, pesticides, and harmful dyes, etc. from wastewater. It has
potential to treat complex effluent obtained from various sources such as domestic wastewater,
dairy wastewater, distillery wastewater, slaughterhouse wastewater, food-processing wastewater,
tannery wastewater, pulp/paper wastewater, and petroleum industry. The energy recovery and
wastewater treatment efficiency from some sources by MFC are shown below [5]:

Figure
SYED6MUHAMMAD
types of MFC ZAKI
with HAIDER
different(BSBT025F18)
sources and their performance 10
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SMFCs for wastewater treatment are applied through a combination of SMFCs and natural
wastewater treatment systems. Biological wastewater treatment mimics microbial degradation of
organic compounds occurring in nature. Thus, natural systems such as water ponds and wetlands
have been used to treat wastewater, sometimes with modification (e.g., aeration). The
combination creates a treatment approach that inherits the advantages of both technologies, such
as energy production in SMFCs and less engineering management with a natural treatment
system [8]. First, wastewater provides more organic compounds (electron source) for anode
oxidation of a SMFC and helps improve electricity generation; second, the electricity-generating
process could stimulate removal of some recalcitrant compounds contained in wastewater or
sediment; and third, the generated electricity may be applied to offset energy consumption by the
treatment system or to power sensors for continuous and automatic monitoring of the treatment
system [8]. Incorporating plants into SMFCs create systems similar to constructed wetlands,
which have been used for wastewater treatment for a long time. Those systems are also called
“CW–MFC” (constructed wetland–microbial fuel cells) in some studies. Plants have a number of
functions in those systems, such as providing substrates for bacterial attachment and biofilm
formation, supplying carbon to microorganisms, uptake of some contaminants (e.g., nutrients),
and regulating water flow in the wetland. There are two major plants used in SMFCs, Ipomoea
aquatica and Phragmites australis, both of which have been applied and/or investigated for
phytoremediation of wastewater in constructed wetlands. It has been demonstrated that the use of
plants could significantly improve electricity generation, and the improvement was likely related
to the increased dissolved oxygen excreted by the plant roots.

Conclusion:
Microbes may look very small but have the potential to do a lot good to our environment when it
comes to cleaning it up, the instance of which is their use in MFCs. MFC is an innovative
technique for power production from wastewater. It is eco-friendly technology, which has shown
excellent pollutant removal efficiency and power generation from simulated as well as real
wastewaters in lab-scale setups. Operating conditions significantly affect the MFC performance
and should be optimized for target wastewater. Low-cost electrodes and membranes are required
to be developed without compromising MFC performance. The integration of MFC with other
technologies/processes (hybrid system) has shown better performance but makes process

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optimization and control difficult. Overall, future research should aim for its operational ease,
cost reduction through economical construction materials, and performance enhancement for its
successful implementation in self-energy sufficient full-scale wastewater treatment.

References:
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light-assisted microbial fuel cells for wastewater treatment and power generation: A win-win
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2. Yaqoob, A. A., Ibrahim, M. N. M., Umar, K., Parveen, T., Ahmad, A., Lokhat, D., &
Setapar, S. H. M. (2021). A glimpse into the microbial fuel cells for wastewater treatment
with energy generation. DESALINATION AND WATER TREATMENT, 214, 379–389.
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3. Microbial Fuel Cells. (n.d.). Let’s Talk Science. Retrieved June 11, 2022, from
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4. Yuliasni, R., Kadier, A., Setianingsih, N. I., Wang, J., Harihastuti, N., & Ma, P.-C. (2021).
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123–144). John Wiley & Sons, Ltd. https://doi.org/10.1002/9781119725008.ch5
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Sustainable Approach for Simultaneous Wastewater Treatment and Energy Recovery.
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https://doi.org/10.1016/j.jwpe.2020.101768
6. FuseSchool - Global Education. (2017). Hydrogen & Fuel Cells | Reactions | Chemistry |
FuseSchool. Retrieved from https://www.youtube.com/watch?v=5_lDGna9MBM
7. Difference between a battery and a fuel cell. (2016, September 20). Watt Fuel Cell.
Retrieved June 11, 2022, from https://www.wattfuelcell.com/difference-between-battery-
fuel-cell/
8. Xu, B., Ge, Z., & He, Z. (2015). Sediment microbial fuel cells for wastewater treatment:
challenges and opportunities. Environmental Science: Water Research & Technology, 1(3),
279–284. https://doi.org/10.1039/C5EW00020C

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9. Konovalova, E. Yu., Stom, D. I., Zhdanova, G. O., Yuriev, D. A., Li, Y., Barbora, L., &
Goswami, P. (2018). The microorganisms used for working in microbial fuel cells (p.
020017). Presented at the INTERNATIONAL CONFERENCE ON ELECTRICAL,
ELECTRONICS, MATERIALS AND APPLIED SCIENCE, Secunderabad, India.
https://doi.org/10.1063/1.5031979

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