Yaqoob 2021

Environmental Technology & Innovation 23 (2021) 101579
Contents lists available at ScienceDirect
Environmental Technology & Innovation

journal homepage: www.elsevier.com/locate/eti
Modern trend of anodes in microbial fuel cells (MFCs): An

overview
∗ ∗
Asim Ali Yaqoob a , , Mohamad Nasir Mohamad Ibrahim a , ,
Claudia Guerrero-Barajas b
a
Materials Technology Research Group (MaTRec), School of Chemical Sciences, Universiti Sains
Malaysia, 11800 Minden, Penang, Malaysia
b
Laboratorio de Biotecnología Ambiental, Departamento de Bioprocesos, Unidad Profesional Interdisciplinaria de Biotecnología,
Instituto Politécnico Nacional, Av. Acueducto s/n, Col. Barrio La Laguna Ticomán, 07340 Mexico City, Mexico
article info a b s t r a c t
Article history: Microbial fuel cells (MFCs) is highly efficient bioelectrochemical systems in which
Received 1 October 2020 bacteria are utilized to oxidize the organic substrate and produce electricity. The main
Received in revised form 25 March 2021 component of an MFCs is the anode, in which bacterial attachment is promoted and
Accepted 18 April 2021
the generation and transfer of electrons occurs. Therefore, the interest of introducing
Available online 21 April 2021
novel and innovative anode materials for attaining a high performance of MFCs is a
Keywords: concern that is growing in the modern era. In this review article, different conventional
Microbial fuel cell and modern anode materials are briefly discussed considering their advantages and
Biomass limitations. Among all, the natural biomass-derived materials were found as one of the
Conventional anode best sources to meet the present challenges to utilize them as anodes in MFCs. The
Graphene anode natural derived anodes presented some limitations which can be addressed through
Composite anode metal oxide composites as described in this article. The oxidation mechanism of
pollutants on the surface of anode along with their bacterial interactions are also briefly
summarized. This review is limited to the performance and development of the anode
in MFCs.
© 2021 Elsevier B.V. All rights reserved.
Contents
1. Introduction............................................................................................................................................................................................... 2
2. MFCs: Mechanisms of energy generation and pollutant remediation ............................................................................................... 3
2.1. Mechanism of electron transfer from bacteria to anode electrode ..................................................................................... 4
2.2. Reduction mechanism of pollutant (Heavy metals) ............................................................................................................... 5
2.3. Biochemical cell reactions and electrochemical measurements ........................................................................................... 6
3. Anode material ........................................................................................................................................................................................ 6
3.1. Conventional anode materials .................................................................................................................................................. 7
3.2. Modern anode material ............................................................................................................................................................. 8
3.2.1. Natural biomass-based anode material .................................................................................................................... 9
3.2.2. Composite-based anode material .............................................................................................................................. 9
3.2.3. Graphene-based anode material ............................................................................................................................... 13
4. Bacterial interaction on anode surface .................................................................................................................................................. 14
5. Economic prospect of natural waste-derived anode electrodes for pilot scale MFCs ..................................................................... 15
6. Problems and future perspective of anode material............................................................................................................................ 15
∗ Corresponding authors.
E-mail addresses: asim.yaqoob@student.usm.my (A.A. Yaqoob), mnm@usm.my (M.N.M. Ibrahim).
https://doi.org/10.1016/j.eti.2021.101579
2352-1864/© 2021 Elsevier B.V. All rights reserved.
A.A. Yaqoob, M.N.M. Ibrahim and C. Guerrero-Barajas Environmental Technology & Innovation 23 (2021) 101579
7. Conclusions................................................................................................................................................................................................ 16
Declaration of competing interest.......................................................................................................................................................... 16
Acknowledgments .................................................................................................................................................................................... 16
Appendix A. Supplementary data........................................................................................................................................................... 16
References ................................................................................................................................................................................................. 17
1. Introduction
The increasing worldwide population, urbanization and industry have led to a very high energy demand. Currently, the
world is getting energy from different fossil fuels sources, but the fossil fuels security, efficiency and their environmental
impact demonstrate that they are not the ideal energy source anymore. For example, in 2011, Japan displayed efforts
to make of nuclear power an alternate energy source to meet the energy crises, but the reliable operation, security and
practical implementation of nuclear power still needs to be safely and completely solved (Mustakeem, 2015). Along with
the high energy demand, environmental pollution is another challenge that has to be taken into account as a consequence
of the energy generation processes, the rising population and the multiple known and unknown natural and anthropogenic
processes carried out to maintain sustenance. Hence, two of the most emerging challenges in modern world are the
increasing energy demand and the management of water pollution, both of which are of upmost importance to maintain
a stable green environment for healthy lifestyle. A vast amount of scientific literature present novel methods to produce
energy and treat wastewater but due to several issues they fail to meet the modern demands (Anthony et al., 2019;
Yaqoob et al., 2020a). Therefore, the development and advance in green energy technologies along with environmental
protection is an essential demand, which currently attracts a wide interest. To address these two challenges, one of the
first alternatives is microbial fuel cells (MFCs), which generate successfully green energy by treating different kind of
pollutants from wastewater. The production of green energy from waste materials by using available electroactive bacteria
such as E. coli, Geobacter spp. etc. is a cost effective and relatively simple process in which by means of bioelectrochemical
processes, carbon is converted to energy (Mian et al., 2019). This process has received a wide interest in the removal of
pollutants from water systems. There are several factors to take into consideration for the MFCs performance, such as
the anode, cathode, organic substrate, and permeable exchange membrane (PEM) (Hindatu et al., 2017; Santoro et al.,
2017). Among all factors, one of the most important is the performance of the anode and the reactions and interactions
that take place in it (Dickhout et al., 2017). The anode electrode is the component of the MFCs in which the bacteria
grow to produce a biofilm around the anode surface. This bacterial growth encourages the decomposition of substrate to
generate protons and electrons. The generated electrons are referred toward the cathodic chamber by using a provided
circuit, whereas the generated protons are transferred directly into the cathodic chamber through a PEM (Gul and Ahmad,
2019). The transferred protons and electrons from the anode are consumed in the cathode chamber. The combination of
protons and oxygen in the cathode chamber, results in the formation of water molecules. The generation of energy through
MFCs depends on the oxidation of organic substrate on the anode surface, whereas the reduction reaction occurs on the
surface of the cathode electrode. Several factors such as extracellular electron transfer (EET), bacterial attachment on
anode, substrate decomposition and biofilm growth are directly linked to the anode electrode (Kaur et al., 2019). Despite
all efforts, the quality of the anode material, cost and free availability issues do not allow the utilization of MFCs at
large scale. An extensive variety of anode materials have been studied as discussed in Section 3 in which they are listed.
The most utilized materials reported for the anode have been carbon-based, metal/metal oxides and conducting polymer
composites. In early time, the precious metals such as gold received significant interest due to their high conductivity
and excellent mechanical strength. Later, to minimize the cost of the anode material, the transitional metals like copper,
nickel, titanium, aluminum, and stainless steel have also been utilized (Yang and Chen, 2020). Similarly, the carbon-based
material such as carbon rod, felt, fiber, graphite etc. are still extensively studied. These are all conventional materials
that have shown several drawbacks such as for example: the metal-based materials showed corrosion under long - term
operation, and high cost, similar conventional carbon material showed low efficiency due to a low electric conductivity
of the material. These conventional materials require extensive surface modification to enhance their quality and thus
minimize their limitations. Recently, Aziz et al. (2020) studied the paper waste — derived, conventional three-dimensional
carbon aerogel, which was further modified with nitrogen-doped reduced graphene oxide (rGO) to enhance the anodic
performance in MFCs. They achieved a 1468 mW/m2 power density. They found that the improvement in conventional
carbon-based material is preferable due to cost effectiveness as well as energy performance. Wang et al. (2020a,b) studied
the conventional carbon foam derived from the corncob wastes. They modified the conventional carbon foam to fabricate
the 3D N-doped carbon foam anode. The prepared 3D N-doped macroporous carbon foam delivered a 4.99 ± 0.02 W/m2 .
The achieved results were quite interesting. Therefore, the modification of the conventional material must be a priority for
high performance in MFCs. It is a good idea to utilize waste material in order to achieve a highly conductive carbon-based
material such as for example, carbon nanotubes (CNTs), carbon black and graphene (Gr‘) derivatives. Later observations
indicated that, among these carbon-based materials, the CNTs and carbon black-based anodes showed toxicity toward the
bacterial community, which may decrease the performance of microbial electrochemical technologies (such as MFCs), as
it was explained by Hassan et al. (2019). Therefore, Gr‘ and its derivatives were introduced as a material that prevents the
extensive limitations that have risen with other carbon-based materials. Gr‘ is a 2D allotrope of carbon which is considered
2
satisfactory for the anode due to its high conductivity, high chemical, thermal, mechanical stability, and biocompatibility
toward bacterial growth. Several previous studies have proved that a carbon anode can significantly encourage bacterial
colonization and formation of a biofilm around the anode surface (Cai et al., 2020). The commercial Gr‘ and its derivatives
are very expensive, therefore, carbon derived from waste biomass material for anode preparation can serve as a low-cost
approach. Several recent studies showed that natural waste derived carbon (which may be converted into precious carbon
form such as Gr‘ derivatives, CNTs etc.) can serve as anode and the reports indicate that significant results were achieved
with them in MFCs (ElMekawy et al., 2017). Additionally, the doping or composite of natural derived precious carbon with
metal oxides can lead to a breakthrough in the field of MFCs. The metal oxides such as ZnO, TiO2 , CuO, AgO, Al2 O3 etc.
have been extensively studied as cost-effective materials because they can be accessible through utilizing natural waste
sources. The composite of these two or more different natural waste materials as anode can serve as an ideal anode in
MFCs. Additionally, the modified waste-derived Gr‘ derivatives anodes have the capability of improving bacteria–anode
interaction, which increases the formation and strength of the biofilm, which in turn may lead to a high electron transfer
from the bacterial cell to the anode electrode. Kumar et al. (2016) considered the preparation of the waste-Gr‘/poly (3,
4ethylenedioxythiophene)/Fe3 O4 nanocomposite as cost-effective oxygen reduction catalyst for high energy generation
and wastewater treatment. The achieved maximum power density was of 3525 mW/m2 . Recently, Yaqoob et al. (2020b)
studied the modification of waste-derived graphene oxide (GO)-based anode to enhance the electron transportation
and remediating power for Pb2+ ions. They used zinc oxide (ZnO) nanoparticle as modifier with waste-derived GO and
achieved a power density of 1350 × 10−3 mW/m2 with a 91.07% removal efficiency of Pb2+ ions. Further, another study of
Yaqoob et al. (2021) described the preparation of cellulose-waste derived GO as anode in MFCs. The maximum achieved
power density was 0.11 mW/m2 . The low power density encourages the modification of the waste-derived anodes for
high performance of MFCs. However, the aim of the present literature review is to discuss the efficiency of energy
generation and MFCs performance by utilization of the modified anode materials. The basic concepts of pollutant oxidation
in the anode and reduction at cathode are briefly discussed. Several conventional as well as modern anode materials
are summarized here for an effective comparison. Moreover, the anode–bacterial interactions are mentioned along with
modern challenges and the possibilities for their utilization in the generation of electricity through the environmentally
friendly approach that the MFCs represent. This review article is limited to the anode materials and their role in MFCs. The
present document is only focused on the transformation mechanism of the heavy metals (frequently present in wastewater
streams) via MFCs with different types of anode.
2. MFCs: Mechanisms of energy generation and pollutant remediation
A fuel cell is usually defined as the conversion of the organic energy into electrical energy without using any kind of
combustion. MFCs approach is also a form of electrochemical fuel cell. MFCs approach promotes the growth of bacteria to
oxidize the organic substrate in wastewater to generate the electrical energy. However, prior to knowing the mechanisms
of pollutant biodegradation or biotransformation through the utilization of MFCs, it is necessary to know the proper
mechanisms of energy generation through MFCs. In the MFCs chambers several strains of bacteria may present the
capability of transferring the electrons and protons through electrodes. It has been found that there are five dominant
groups of microorganisms such as Firmicutes, Proteobacteria, Acidobacteria, fungi and algae that present the characteristic
of electricity generation associated to their respiration process in the MFCs chamber (Kumar et al., 2015; Yaqoob et al.,
2020c). Some bacteria that have been reported earlier as electron exchanger with electrodes, are Clostridium butyricum,
Rhodoferax ferrireducens, Shewanella sp., Geobacter spp. and Aeromonas hydrophila (Chandrasekhar, 2019; Guizani et al.,
2019; Jung and Pandit, 2019; Michelson et al., 2019). Furthermore, there are some bacterial species that can transfer the
electrons directly to the anode without transference to an exogenous acceptor. The bacterial species that carry out this
process are known as exoelectrogens (He, 2017).
In MFCs, the bacterial species break down different organic compounds and heavy metals complexes to produce
electrons and protons in order to empower their respiration system while exhibiting the capability of producing a flow
of electrons by means of the electrodes. However, microorganisms can transfer electrons into electron acceptors that
are present in an insoluble state. For example, some Geobacter sp. have pili, which are as conductive as metal and are
actively expressed. Microorganisms grow on the surface of the electrodes and develop a biofilm to transfer electrons more
efficiently than through the utilization of insoluble electron acceptors (Yang et al., 2019). The formation of biofilm on the
surface of the anode is the most significant type of interaction to transfer the electrons. To minimize the competition
between redox mediators and oxygen, anaerobic conditions must be applied. The oxygen supply (oxygen is an electron
acceptor) in the anodic chamber leads to poor energy generation (Nosek et al., 2020). So far, in regard to the biofilm,
both types of bacterial cultures have been reported in literature, bacterial consortia and pure bacteria cultures (single
bacterial species). According to the Jadhav and Ghangrekar (2020), the mixed consortia offer higher power efficiency
than pure cultures. The key advantage of the bacterial consortia is that they can oxidize the organic substrate more
effectively. This is due to the syntrophic dynamic between exoelectrogens and fermentative bacteria, such interaction
may improve the exoelectrogenic activities. On the other hand, the pure single bacterial colonies are easy to investigate
in the biofilm in regard to the electron transfer mechanisms. However, the anaerobic atmosphere in the anodic chamber is
an essential condition to carry out the metabolic pathway for the oxidation of the complex organic substrate by bacterial
species. During the oxidation process, any complex organic substrate is first hydrolyzed into simple compounds such as
aromatic compounds, monosaccharides, fatty acids, and amino acids as shown in Fig. 1. After that, the hydrolyzed simple
compounds are fermented or oxidized to CO2 , which may generate and transfer the electrons to the anode electrode. The
maximum generation of electrons and complete oxidation of simple compounds is an ideal condition in MFCs.
3
Fig. 1. Oxidation process of organic substrate on the anode surface in the presence of a biofilm to generate the electron and proton.
Source: Reproduced from Nosek et al. (2020) with MDPI permission.
2.1. Mechanism of electron transfer from bacteria to anode electrode
Microorganisms that transfer the electrons extracellularly are classified as exoelectrogens and some species that
have the ability to transfer electrons in this way are: Geobacter lovleyi, Geothrix fermentans, Thermincola carboxydophila,
Geobacter sulfurreducens, Shewanella oneidensis, Rhodopseudomonas palustris, Thermincola potens, E. coli, and Shewanella
putrefaciens (Abbas et al., 2017; Anand et al., 2019; Verma et al., 2015). For current generation, exoelectrogens can transfer
electrons from electrodes through some mechanisms that have been clearly stated such as short-range electron transfer via
shuttling molecules (also known as redox mediators), electron transfer through redox-active proteins, long-range electron
transport by conductive pili and direct interspecies electron transfer as shown in Fig. 2.
i. In brief, the short-range transfer of electrons toward the anode electrode is carried out via electron shuttles
molecules such as in the case of G. fermentans. Both, gram positive or negative bacteria, can transfer electrons
through using self-producing shuttle molecules. Desulfuromonadaceae and Geobacteraceae are two families of
bacteria that produce self-electron shuttles, for example, c-type cytochromes are a group of these electron shuttles
that include MtrC, MtrD, MtrF, OmcA and MtrE components (Abbas et al., 2017; Saratale et al., 2017).
ii. Geobacter sulfurreducens is a good example of microorganism that can assist the electrons transformation toward
the anode via anaerobic enzymatic-based metabolism activities. The bacteria Geobacter sulfurreducens facilitates the
electron transformation to several types of acceptors such as fumarates etc. It has been reported that several redox
active proteins such as OmcT, OmcZ, OmcS, OmcE and OmcB are available in exoelectrogens (Sevda et al., 2018).
iii. The long-range transfer of electrons through conductive pili attracts great interest. A little filamentous projection in
bacteria body, which promotes surface adhesion and is not used for mobility is known as pili. These pili are usually
conductive and help to transfer the electrons from the biofilm present around the anode surface. The conductive
pili serve as nanowires to transfer the electrons toward the anode. The conductive pili are generally available in
the strains of Shewanella oneidensis, Pelotomaculum thermopropionicum, G. sulfurreducens, and Methanothermobacter
thermautotrophicus (Zhou et al., 2017a,b,c). The direct interspecies electron transfer approach was studied mostly in
Geobacter Metallireducens and G. sulfurreducens. In this approach, the microorganism serves to transfer the electron
directly and enhances the mutual growth. P. thermopropionicum and Synechocystis are also an example of this kind
of mechanism in MFCs (Abbas et al., 2017; Shi et al., 2019).
4
Fig. 2. Schematic presentation of MFCs and electron transfer mechanisms from exoelectrogens to electrodes. (i) Short range electron transfer (ii)
Redox active proteins (iii) Long range transfer via conductive pili.
2.2. Reduction mechanism of pollutant (Heavy metals)
To date, several chemical, physical, analytical, and biological approaches have been introduced for the remediation of
the heavy metals. Among all, MFCs is the feasible, cost-effective, and eco-friendly approach, which can remove the heavy
metals with simultaneous generation of energy. The biological mechanisms are very significant in remediation of heavy
metals in MFCs. The microorganism properties to accept electrons from (anode and cathode) electrodes are known as
electrotrophs (Xionga et al., 2019). This fact gives a new direction for treatment of heavy metals via reduction. The heavy
metals are removed through the reduction reaction at cathode while organic substrates are oxidized at anode and serve
as electron donor and carbon source to the microorganisms (Ucar et al., 2017).
There are many types of bacteria with the property of gaining electrons directly from electrodes. Recently, many studies
have reported that Geobacter species can accept electrons directly from electrodes (Saratale et al., 2017). There are many
toxic heavy metals like chromium ion, Ni, Zn, Hg, Pb, Cu, V etc. that are reduced by different microorganisms by following
the same mechanisms, i.e., G. sulfurreducens accepts electrons directly from electrodes and reduces the U(VI) into U(IV)
form (soluble to insoluble). The U (IV) is insoluble, and it is adsorbed on the electrodes (Tizaoui et al., 2019). Watts
et al. (2015) studied G. sulfurreducens that has the capability of reducing Cr (VI) to Cr (III), it can convert the highly toxic
oxidation state of chromium to the less toxic form. The reduction of Cr (VI) depends on the oxidation of substrate (acetate)
at the anode electrode to transfer the electrons to microorganisms and the reduction of chromium occurs at the cathode.
The reduction reaction can be written as:
Cr2 O7 −2 + 14H+ + 6e− −→ 2Cr3+ + 7H2 O
Hao et al. (2015) studied the removal of vanadium with microorganisms such as Enterobacter, Macellibacteroides and
Lactococcus, these authors demonstrated that vanadium removal presented a 93.6% efficiency and a high current density
of 543.4 mW/m2 . However, the most toxic heavy metal mercury can be reduced via MFCs. The Hg2+ redox potential is
−320 mV, it can be taken as electron acceptor. The removal mechanism of Hg2+ in precipitate form occurs in the presence
of chloride (Cl− ) ions and the reduction through electrons occurs at the cathode electrode as shown below:
2Hg2+ + 2Cl− −→ Hg2 Cl2 (s)
Hg2 Cl2 (s) + 2e− −→ 2Hg(l) + 2Cl−

The final product Hg2 Cl2 is deposited at the bottom of the cathode and elemental Hg on the surface of cathode. During
this process, the achieved maximum power density was of 433.1 mW/m2 (Wang et al., 2011).
Similarly, the reduction mechanism of Cu2+ to Cu can be written as:
Cu2+ + 2e− −→ Cu
2Cu2+ + H2 O + 2e− −→ Cu2 O + 2H+

Cu2 O + 2e− + 2H+ −→ 2Cu + H2 O
5
As a result of the Cu2+ reduction in MFCs operation, two major products may be formed at the cathode: Cu2 O or Cu,
whereas the electrons come from the oxidation process of organic substrate that occurs at the anode compartment
(Fang and Achal, 2019). After an extensive review, we found that there is no proper molecular mechanism known to
describe how the microorganisms can accept electrons from electrodes. This is a very useful direction for research in the
future. However, in the whole investigation, the anode has received significant attention in the MFCs research because
the working mechanism of MFCs indirectly depends on the performance of the anode. Therefore, the improvement of the
anodic component of these systems is of upmost importance over others. Some recent studies related to the removal of
toxic heavy metals in MFCs are shown in Table S1.
2.3. Biochemical cell reactions and electrochemical measurements
During the MFCs operation, the organic substrates such as glucose, acetate sucrose etc. are oxidized by bacteria to
generate the electrons. The oxidation reaction is carried out at the anode and the reduction reaction at the cathode, some
examples of these reactions can be written as follows with different organic substrates.
(a) If glucose is used as organic substrate

Oxidation reaction at anode: C6 H12 O6 + 6H2 O −→ 6CO2 + 24H+ +24e−
Reduction reaction at cathode: 24H+ + 24e− + 6O2 −→ 12H2 O
Overall reaction: C6 H12 O6 + 6O2 −→ 6CO2 + 6H2 O + Electricity + Biomass
(b) If acetate is used as organic substrate
Oxidation reaction at anode: CH3 COOH + 2H2 O −→ 2CO2 + 8H+ +8e−
Reduction reaction at cathode: 8H+ + 8e− + 2O2 −→ 4H2 O
Overall reaction: CH3 COOH + 2O2 −→ 2CO2 + 2H2 O + Electricity + Biomass
(c) If sucrose is used as organic substrate
Oxidation reaction at anode: C12 H22 O11 + 13H2 O −→ 12CO2 + 48H+ +48e−
Reduction reaction at cathode: 4H+ + 4e− + O2 −→ 2H2 O
Overall reaction: C12 H22 O11 + 12O2 −→ 12CO2 + 11H2 O + Electricity + Biomass
Furthermore, the produced electrons move from anode to cathode electrode while different electrochemical measure-
ments are carried out to assess the performance of the electrode. The voltage potential trend can be measured by using
the digital multimeter. To calculate the voltage in form of current (Ampere), Ohm’s law is followed. The power density
(P.D), current density (C.D) and internal resistance (r) can be calculated by using the following Equations:
V = IR (1)
2
V
P.D = (2)
RA
I
J= (3)
A
E−V
r=[ ]R (4)
V
where, I = current, V = voltage output, R = external resistance, r = internal resistance, A = cross-sectional area, J =
current density (CD), E = Electromotive force (Emf). The Emf can be obtained via open circuit voltage through connecting
to resistance voltmeter. Additionally, the cyclic voltammetry (CV), specific capacitance (Cp ) and electrochemical impedance
spectroscopy (EIS) are also carried out to investigate the electrochemical performance of the electrodes. The Cp is carried
out to study the power of the biofilm in order to transfer the electrons. The Cp can be calculated by Eq. (5).
AV
Cp = (5)
2mk(V2 − V1)
where, AV = CV curves area, m = sample amount in CV instrument, k = scan rate (mV/s), (V 2 − V 1) corresponds to the
total voltage range in CV.
3. Anode material
The anode material is significant in MFCs because the anode provides the environment in which the oxidation process
(of organic compounds) is carried out, whereas the cathode helps to activate the reduction process. Out of the two
electrodes, the anode improvement is more essential for a better performance of a MFCs compared to the cathode.
ElMekawy et al. (2017) studied the comparison between the anode and cathode performance in MFCs and proved that the
anode materials improvement is more necessary than the cathode materials as is shown in Fig. S1. They used Gr‘-based
material as anode and cathode in separate cells and achieved 12.5% of energy generation in the case of Gr‘anode over
the Gr‘cathode. However, the selection criteria of anode materials are quite difficult due to several issues such as cost,
long – term stability of anode etc. The anode material choice exerts a direct influence on other MFCs parameters such as
6
substrate oxidation, electron transformation, bacterial adhesion, biofilm characteristics, biofilm structure, and growth of
bacteria (Khudzari et al., 2019). The other parameters of the MFCs are also very important but they all directly depend on
the performance of the anode material. Therefore, to achieve a high energy output, the anode material should be ideal.
Although the selection criteria of cathode are quite different from those for the anode. Therefore, after a wide literature
review, some anodic properties have been identified as responsible for achieving a high performance of MFCs, they are
listed as follows (Gajda et al., 2020; Saadi et al., 2020):
i. Material surface area: The energy generation through MFCs is seriously inhibited by the material of the surface
area of the anode. The electrode resistances are directly proportional to ohmic losses. Therefore, an increase of a
proper material on the surface area, for example, by utilizing modern materials or modified materials to minimize
the resistance is desirable to attain better results. A higher surface area provides a more active site for the reactions
to occur, which enhances the electron generation and movement. All biological and electrochemical reactions occur
on the anode electrode; therefore, the surface area tremendously influences the MFCs performance. Gonzalez et al.
(2019) tested two different surface area-based electrode materials in the presence of microalgae Scenedesmus
obliquus as biocatalyst in MFCs operation. They concluded that the graphite electrode with larger surface area
(13.6 cm2 ), rather than a smaller surface area (5 cm2 ), delivered a higher energy output. The 13.6 cm2 based graphite
rod yielded 109 ± 2 µA, whereas other yielded 37 ± 5 µA. They proved that the larger surface area of the anode
electrode exerted a major impact on MFCs performance.
ii. Biocompatibility of material: The bacteria are used to form a biofilm around the anode surface and oxidize the
organic substrate. The material should be biological favorable, chemically stable, and toxic free for healthy bacterial
growth. A toxic nature of the anode material inhibits the healthy growth and respiration of bacteria which in turn
fails in the generation of protons and electrons as a result. Different characterizations such as SEM images, EDX and
chemical toxicity tests are carried out to ensure the biocompatibility of the material. The modern advanced electrode
material should be highly biocompatible. For example, a green synthesized metal oxide with carbon-based material
nanocomposite can effectively minimize the toxicity and improve the interaction between bacteria and electrode.
iii. Electrical conductivity: The material conductivity properties are essential because electrons are traveling from
anode to cathode, which requires a conductive anodic material. The highly conductive anode material promotes the
flow of electrons better and this minimizes the resistance of the electrode and leads to a good outcome. Kirubaharan
et al. (2019) modified the conventional carbon cloth electrode by using the Au@PANI composite to improve the
conductivity of the anode material. The aim of that study was to improve the electron transfer rate from anode to
cathode. The modified carbon cloth delivered 804 ± 73 mW/m2 energy efficiency, which is 3 times higher than the
obtained with an unmodified carbon cloth anode. Therefore, the material conductivity is one of the most significant
parts in order to achieve high performance.
iv. Material stability and durability: The long-term stability and durability is still an issue in modern research since
the anode stability is a key factor in achieving the goal of a sustainable MFCs. The oxidizing environment on the
surface of the anode leads to corrosion, swelling and decomposition of the materials. These issues cause fouling,
which minimizes the anode stability for a long-term operation of MFCs. Senthilkumar et al. (2019) studied the
neem wood waste derived biochar which was modified with PEDOT/NiFe2 O4 to enhance the material properties
particularly durability and stability. They reported the 350 h durability of the prepared modified anode. Additionally,
in a recent research document (Yaqoob et al., 2020c) reported a 90-day durability and stability for graphene derived
modified anode in MFCs. Thus, the anode material must be mechanical, chemical, thermally stable, and durable in
both, acidic and basic conditions in the MFCs.
v. Material availability and cost: To implement the MFCs systems at large scale, the material availability and its cost
must be feasible. For a commercial setup, the MFCs electrode materials must be easily available at low cost. For
example, gold, platinum, silver etc. are very expensive and they are not available everywhere. The composites anode
might be another way to meet this criterion for large scale applications of MFCs in the future.
Recently, ongoing investigations are focused on the development of anode materials to improve the MFCs approach to
an industrial level. However, some progress and development for the anode configurations have been considered in this
article, particularly in two broad types, the so-called conventional anode materials, and the modern anode materials.
3.1. Conventional anode materials
In previous decades, many studies reported on the development of anode materials and some conventional materials
such as some carbon-based, metal/metal oxides were the most familiar as is shown in Table S2. Over time, these
conventional materials have been replaced with some new modern materials, which showed better results in MFCs. In a
conventional anode, the carbon-based is the most familiar material, even later carbon-based modified materials became
a modern material for anode. The carbon-based materials gained significant attention due to their high conductivity,
biocompatibility, and their chemical/mechanical stability. The conventional carbon-based materials include carbon paper
(CP), felt (CF), rod (CR), cloth sheet (CS), brush (CB), foams (CFO), graphite plate (GP), rod (GR), sheet (GS), cloth
(GC), graphite granules (GG), activated carbon (AC) and reticulated glassy carbon (RGC), respectively, as is presented in
7
Fig. 3. Some of the most conventional anode materials utilized for the anode (a) CP (b) CC (c) CF (d) RGC (e) CM (f) GG (g) CBs (h) GR (i)
polycrystalline graphite (j) CF (k) Pt mesh (l) few metallic anode strips (m) few conducting polymer anode strips.
Source: Reproduced from Ref. Wei et al. (2011) with Elsevier permission.
Fig. 3. Further, the packed and plane structural configuration and brushes-based conventional anodes have already been
extensively reviewed by Wei et al. (2011). All mentioned carbon-based materials are and have been studied in MFCs
based on a specific projected area, this may be a surface area that has not been well delimited in order to adequately
assess the entire MFCs performance. For example, He et al. (2017) demonstrated that the calculation of actual surface
area is quite difficult, especially when a MFCs was operated with a porous anode. This leads to a significant variation in
exploiting the power density, which varies greatly in systems working with different porous anodes in the same surface
area. Therefore, it is essential to introduce a suitable benchmark to precisely analyze the performance of several anode
materials in MFCs. There are some aspects that limit the applications of these conventional carbon materials. For example,
the graphite-based materials present a better mechanical stability, but a relative lower biofilm formation on the anode
surface, which directly affects the energy generation efficiency in MFCs. The CP, CC and RGC anode offer significant rough
surface area, which is good for biofilm formation, but these all are mechanically poor; therefore, they cannot be used for
long term operations. The CF showed excellent conductivity and flexibility toward the bacteria adhesion. The production of
the biofilm limits the diffusion process of organic substrate from the exterior to the interior surface, which is unfavorable
for healthy bacterial colonization. This is due to the high thickness surface of CF material. In contrast to others, the CBs
have improved performance in energy output, but metal wires are required to fix the carbon brushes, which makes it
economically unsuitable. Therefore, the great majority of researchers are using CP, CF, or GR to develop the modified anode
(Sonawane et al., 2017). Thus, it is not easy to fulfill the requirements for an ideal anode in practical MFCs applications.
Similarly, several types of metal/metal oxide nanoparticles (NPs) such as MnO2 (Phonsa et al., 2018; Xu et al., 2018),
Au (Jarosz et al., 2019), WO3 (Varanasi et al., 2016), TiO2 (Ying et al., 2018), Pd (Quan et al., 2018), and several others
(Mohamed et al., 2017; Gao et al., 2018; Taşkan et al., 2019) have been tested. The idea of using metal-based anode
strips has failed due to several reasons such as cost, corrosion, non-biocompatibility etc. Taşkan et al. (2019). For example,
stainless steel mesh is one material that fulfills several requirements of an ideal anode, but biofilm vanishes in long – term
operation due to the gravity effect (Leng et al., 2020). Similarly, Au, Ag, and Cu also exhibit excellent conductivity, but
these all showed low operational stability in long – term periods to generate the energy. Therefore, to obtain a satisfactory
MFCs performance, modern anode materials must be introduced to the design while looking for feasible ways to acquire
them.
3.2. Modern anode material
The modern anode materials are further sub-classified into two categories, which have exhibited their significant
importance in generation of energy as well as in wastewater treatment concerns. The subcategories of modern anode
materials are natural biomass-based carbon anode material and composite-based material that includes metal/metal
oxides, carbon-based materials and conducting polymers (Slate et al., 2019). The modern materials are considered as
a low-cost alternative for the anode over a conventional material and with the advantage of having excellent properties.
Several studies have explained the value of modern anode materials that have been used in MFCs. A description of these
modern materials is included in the following sub-sections.
8
3.2.1. Natural biomass-based anode material

The natural recyclable sources provide a potential green method for producing the valuable bioenergy from natural
waste and sustainability. The plant waste is the most frequently utilized natural precursor for carbon-based anodes. This
is commonly known as natural waste-derived anode material. However, the porous carbon material is generally achieved
through direct carbonization of plant precursors at very high temperatures (1050 ◦ C). In the preparation process, the plant
heteroatoms act as natural dopants, subsequently as self-doped material, whereas the water evaporation throughout the
carbonization (at 1050 ◦ C) leads to the development of a porous structure in the carbon material (Antolini, 2016). Zhang
et al. (2018) synthesized the 3D microporous material by using bread pieces via direct carbonization process in presence of
self-doped carbon foam (N, P and S). The resultant product showed high porosity along with 295.07 m2 /g of surface area,
high conductivity, and excellent dopants (N, P, S) distribution as shown in Fig. S2 (a-d). This kind of natural-synthesized
anode produced up to 3134 mW/m2 , which is three-times higher than the results obtained with a CC anode.
Recently, Hung et al. (2019) studied the natural waste derived porous carbon anode and achieved a 3927 mW/m2
power density, which is higher than the obtained with an activated carbon anode. The improvement on the power density
of coffee-waste derived anode was due to a high electrical conductivity and appropriate pore size, which promote the
electron movement and bacterial colonization on the anode. Additionally, the long-term MFCs performance with coffee-
derived anode was also examined, it unceasingly functioned for 100 h, with the obtainment of power density (2000
mW/m2 ) without any additional nutrient. These outcomes demonstrate that coffee-waste derived anode is excellent for
energy generation that could dramatically decrease the entire cost of MFCs. Former reports indicate that many natural
sources have been employed for synthesis and fabrication of anodes for MFCs as is summarized in Table 1. These all
natural-derived anodes have high active surface, available surface area, and their porosities are considered as leading
factors that influence the MFCs performance. Chen et al. (2016) produced a hierarchical designed anode by utilizing the
chestnut shells waste, which served as a precursor. The brush structure of spherical shell offered an active and efficient
surface for biofilm growth on the anode. The achieved power density was of 759 ± 38 mW/m2 with a 75% of coulombic
efficiency, which are usually comparable with values obtained with conventional GB anodes as is shown in Fig. 4(a,b). One
more study described the chestnut shell-based anode material with chemical activation and obtained the highest power
density, which is higher than the obtained with the conventional CC anode (Chen et al., 2018). However, to improve the
bacteria loading sites without raising the weight and volume of the anode material Zhu et al. (2014) synthesized the
hollow structure fibers material and used them as an anode. This hollow structure offers an opportunity to transfer the
electrons in both surfaces (inside and outside) as is shown in Fig. 4c. This type of anode produced 20 times higher power
density than the obtained with the conventional CF electrode. Further improvement has been shown in the case of the
carbonization mechanism and modest natural precursors coating with carbon nanomaterial, which are usually some of
the energy-saving methods in anode fabrication. Zhou et al. (2017a,b,c) fabricated the 3D anode through a soaked loofah
sponge with ink coating, which showed extraordinary elasticity, and an exposed porous structure in the material. One
important observation is that this type of anode showed slightly better mechanical stability than the conventional under
70 days of continuous operation and produced (16.3 ± 0.5 mA/cm3 ) a high current density. Similarly, another research
group, Zheng et al. (2016a) achieved a 61.7 ± 0.6 W/m3 power density with soaked loofah sponge 3D anode in MFCs.
Therefore, it is proposed that the soaking approach can act as generic, low-cost strategy for the preparation of the anode.
Lu et al. (2016) prepared a bioanode by simple pyrolysis of silk cocoon at very high temperature, which shows
hierarchical pores as presented in Fig. S3 (a, b). The fabricated elastic carbon fiber anode exhibited high mechanical
stability and produced a supreme power density, which is usually higher than the obtained in the CC anode. According to
the discussion, the carbonization of different biomass at very high temperature is a general method for synthesis of anode
material. The biomass can be converted into porous-structure derived carbon through introduction of heteroatom. This is
the most promising cost-effective method and source that could serve at commercial level. However, the natural anode
porosity is a factor that exerts a direct influence on bacteria colonization by disturbing the surface area of the anode.
The normal size of bacteria is usually 0.4–4 µm and typically 1–2 µm respectively. Therefore, 2–10 µm is a preferable
pore size for bacteria cell colonization (Chong et al., 2019). Although the pore size of biomass-based anode is mentioned
mostly for the rare precursors, in general, further processing such as chemical activation is required to control the pore
size distribution during the fabrication of the anode.
3.2.2. Composite-based anode material

The different material composite-based anodes have received significant attention in current years. The composite-
based materials have been usually utilized to attain the synergistic influence with more than two types of materials or
amendments in base materials, which leads to highly upgraded anode kinetics efficiency. The Table 2 summarized the
performance of MFCs in the presence of composite-based anode, which demonstrated the high efficiency of energy gener-
ation as compared to unmodified anode material (Cai et al., 2020). The composite-based anode material is categorized into
different classes such as metal/metal oxides, conductive polymers, and carbon-based composites with each other. In fact,
the improved bacterial adhesion and decrement in ohmic losses are greatly influenced by metal/metal oxide- carbon-
based material composites (Cai et al., 2019; Luo et al., 2019). The metal oxide/carbon composite gained great interest
due to several reasons because metal-based material alone is facing a corrosion problem and carbon-based are suffering
of lower conductivity. Therefore, the composite addresses these two factors and offers a perfect model anode for MFCs
performance. Tang et al. (2015) used natural egg white protein (EWP) waste to prepare a carbon anode, modified it with
9
Fig. 4. (a) Graphic mechanism of chestnut shell anode preparation. (b) images of GB and chestnut shell-based anode (reproduced from Chen et al.
(2016) with Elsevier permission). (c) hollow structure fibers-based material (reproduced from Zhu et al. (2014) with Elsevier permission).
Fig. 5. (a) A graphic sequence for the design of Loofah sponge fiber-TiO2 -based carbon electrode (b) images of loofah sponge, TiO2 /EWP carbonized
carbon, and loofah sponge carbon-TiO2 composite. (reproduced from Tang et al., 2015 with Elsevier permission) (c, d) SEM image of SSM/TiO2
(reproduced from Ying et al., 2018 with Elsevier permission) (e) SEM image of NiO/PANI/CF (reproduced from Zhong et al., 2018 with Elsevier
permission) (f, g) TEM image of Ni0.1Mn0.9O1.45 product. (reproduced from Zeng et al., 2018b with Elsevier permission).
TiO2 (titanium oxide) and offered a 3D nano-structured modified anode for MFCs. This was combined with loofah sponge
carbon to achieve the 3D anode electrode by carbonization process, as is shown in Fig. 5a. The TiO2 coating on loofah
sponge carbon and the thermal treatment lead to the nano size particles as is presented in Fig. 5b. This type of modified
composite anode offered a high 200% energy generation efficiency, higher than the obtained with a conventional graphite
anode. The increased energy output was attributed to the increased electro-chemical power of the 3D based anode. The
natural EWP waste utilized to derive a carbon anode exhibits great surface area, promising surface functionalization, high
rate of extracellular electron transformation and high biocompatibility.
In regard to CNTs, former research showed that CNTs conveyed biotoxicity toward living cells, which might confine
the bacterial proliferation and execute the cells (Yazdi et al., 2016). The CNTs material applications in MFCs as anode
was in question. It required proper modification with specific functional groups to decrease the biotoxicity toward cells.
Amazingly, some proper modifications showed optimistic effects on refining the efficiency of MFCs. For instance, Zhang
et al. (2017a) developed an idea of CNTs hybrid biofilm formation through contact with (multi-walled carbon nanotubes)
10
Table 1
List of recently reported biomass-derived anodes.
Biomass-derived Synthesis method Organic substrate Source of Power density References
anodes sources inoculum
Loofah sponge Carbonization and Acetate Pre-acclimated 1090 ± 72 Yuan et al. (2013)
polymerization bacteria mW/m2
Bamboo charcoal Simple Acetate Pre-acclimated 1652 ± 18 Zhang et al. (2014)
carbonization bacteria mW/m2
Kapok fibers Carbonization Acetate Activated sludge 1738.1 mW/m2 Zhu et al. (2014)
King mushroom Carbonization Acetate Anaerobic digester 2.09 mW/cm2 Karthikeyan et al. (2015)
sludge
Wild mushroom Carbonization Acetate Anaerobic digester 3.02 mW/cm2 Karthikeyan et al. (2015)
sludge
Corn stem Carbonization Acetate Anaerobic digester 3.12 mW/cm2 Karthikeyan et al. (2015)
sludge
Coconut shell Carbonization Acetate Sludge 1069 ± 15 (Yuan et al., 2015)
mW/m2
Chestnut shell Carbonization and Acetate Pre-acclimated 759 ± 38 mW/m2 Chen et al. (2016)
activation bacteria
Loofah sponges Carbon Acetate Pre-acclimated 61.7 ± 0.6 W/m3 Zheng et al. (2016b)
black-coating bacteria
Silk cocoon Simple Acetate Activated 5.0 mW/m2 Lu et al. (2016)
carbonization anaerobic sludge
Municipal sludge Simple Lactate Shewanella 568.5 mW/m2 Ma et al. (2016)
carbonization oneidensis MR-1
Sugarcane Carbonization Acetate – 59.94 ± 2.81 Zhou et al. (2017a,b,c)
W/m3
Loofah sponges Chines black coating Acetate Pre-acclimated 0.82 mW/cm3 Zhou et al. (2017a,b,c)
bacteria
Sewage sludge Chemical vapor Acetate Pre-acclimated 2228 mW/m2 Feng et al. (2018)
method bacteria
Cake Carbonization and Acetate Pre-acclimated 1307 mW/m2 Yuan et al. (2018a)
polymerization bacteria
Cotton textile Carbonization and Acetate Activated sludge 931 ± 61 mW/m2 Zeng et al. (2018a)
polymerization
Chestnut shell Carbonization and Acetate Municipal waste 23.6 W/m3 Chen et al. (2018)
activation
Sewage sludge Carbonization Acetate Pre-acclimated 3.2 W/m2 Jia et al. (2018)
bacteria
Bread Simple Acetate Pre-acclimated 134 mW/m2 Zhang et al. (2018)
carbonization bacteria
Neem wood Simple Simple glucose Pre-acclimated 256 ± 25 mW/m2 Senthilkumar et al. (2019)
Cocklebur fruit Simple Acetate Pre-acclimated 572.57 ± 24.90 Yai et al. (2019)
carbonization bacteria µW/m2
Pinecone Simple Acetate Pre-acclimated 10.88 W/m3 Wang et al. (2019)
Coffee Carbonization Acetate Domestic waste 3927 mW/m2 Hung et al. (2019)
(continued on next page)
MWCNT in the presence of the biofilm. It improved the composite conductivity of the biofilm, increased the rate of
electron transfer as well as the organic substrate diffusion. Furthermore, the starting period was also decreased, and
composite biofilm anode showed a long-time stability and strength. These factors led to excellent performance of MFCs.
Undeniably, further improvements can be carried out through CNTs doping with conductivity polymers to enhance the
MFCs efficiency (Cai et al., 2020). Cui et al. (2015) produced a CNTs/GF/PANI-based composite and used it as anode. The
preparation of CNTs/GF/PANI was carried out by polymerizing the PANI on GF, which was further treated with CNT via
electrophoretic deposition. This composition enhanced the material electrical conductivity, hydrophilicity, and surface
roughness, respectively. The achieved power density was of 257 mW/cm2 , which is 3.4 times higher than the obtained
with a GF/PANI composite-based anode. The discussion showed that the modified CNTs anode material can increase the
electrocatalytic conductivity of the material and significantly enhance the surface area of it. Therefore, the surface of anode
11
Table 1 (continued).
Oil palm empty fruit Carbonization Oil palm trunk Pb2+ 1350 × 10−3 Yaqoob et al. (2020b)
bunch waste sap supplemented mW/m2
pond wastewater
Oil palm empty fruit Carbonization and Oil palm trunk Pb2+ 20 × 10−3 Yaqoob et al. (2020b)
bunch waste modification sap supplemented mW/m2
pond wastewater
Corncob wastes Carbonization and Acetate Starch wastewater 4990 mW/m2 Wang et al. (2020a,b)
modification
Cellulose-wastes Carbonization and Sweet potato Synthetic 0.11 mW/m2 Yaqoob et al. (2021)
modification waste wastewater
Table 2
List of composite material used as anode materials in the last few years.
Composite anode material Surface Substrate Source of inoculum Power Reference
area density
(cm2 ) (mW/m2 )
CNFs/GF 1 Acetate Wastewater – Shen et al. (2014)
SnO2 /rGO/CC 6 Phosphate buffer/glucose Escherichia coli 1624 Mehdinia et al. (2014)
TiO2 –CC nanofiber mats – Luria Bertani broth E. coli 8 Garcia-Gomez et al. (2015)
Mobil catalytic materials/GF – Synthetic wastewater Anaerobic digester sludge 71.8 Wu et al. (2015)
MnO2 /CF 7.1 Phosphate buffer/acetate MFCs effluent 3580 Zhang et al. (2015)
Pd/CC 10 Phosphate buffer/acetate S. oneidensis MR-1 500 Quan et al. (2015)
PANI/Gr-CC 7 Phosphate buffer/acetate MFCs effluent 884 Huang et al. (2016)
FeO/CP 12 Acetate Strain breeding 140.5 Harshiny et al. (2017)
Mo2 C/CF 2 Lactate S. putrefaciens CN32 1025 Zou et al. (2017)
Co/CoxOy/Graphite 6.25 Phosphate buffer/acetate Food wastewater 576 Mohamed et al. (2017)
PANI/CC 4 Phosphate buffer/acetate MFCs effluent 567 Zhang et al. (2017b)
TiO2 /SSM 7 Acetate MFCs effluent 2870 Ying et al. (2018)
Fe3 O4 /CC 6.25 Simulated wastewater Activated sludge 728 Xu et al. (2018)
NiO/PANI/CF 9 Phosphate buffer/acetate Anaerobic sludge 1079 Zhong et al. (2018)
Ni0.1Mn0.9O1.45/CF 4 Acetate MFCs waste 1390 Zeng et al. (2018b)
PANI/SS plates 3.5 Phosphate buffer/acetate Anaerobic mixed sludge 780 Sonawane et al. (2018a)
PANI/SSW 7.5 Phosphate buffer/acetate Anaerobic mixed sludge 2880 Sonawane et al. (2018b)
Polypyyrole (Ppy)/SSW 7.5 Phosphate buffer/acetate Mixed sludge 1870 Sonawane et al. (2018b)
Ppy/SS 4 Phosphate buffer/acetate Anaerobic granular mixed sludge 1191 Pu et al. (2018)
Ppy/SS 4 Phosphate buffer/acetate Anaerobic sludge 440 Phonsa et al. (2018)
Fe3 O4 /porous graphitized 4 Phosphate buffer/acetate MFCs effluent 1856 Hu et al. (2019)
carbon
Carbon microwires/titanium 2 Acetate MFCs effluent 12.2 Ying et al. (2019)
meshes
NiWO4 /GO/CC 1 Simulated wastewater Anaerobic mixed sludge 14 581 Geetanjali et al. (2019)
PPy/MnO2 /SS 50 Simulated wastewater Mixed anaerobic sludge 1877 Zhao et al. (2019)
Carbon thread brush/titanium 164 Acetate S. frigidimarina 2.42 Vilela et al. (2020)
Ppy-carboxymethyl cellulose- – Acetate Mix culture 2970 Wang et al. (2020a,b)
CNTs-CB composite anode
Activated Carbon-Polyaniline 50 Synthetic wastewater Anaerobic bacteria consortia 273 Yellappa et al. (2020)
composite
offers more active sites for healthy bacterial growth/colonization and promotes the substrate transformation mechanism
entirely (Zhao et al., 2017). However, Delord et al. (2017) studied the preparation of CNTs fiber mats-based anode for MFCs
through interlacing fibers containing exclusively the CNTs. The prepared anode presented an enormously porous structure
as well as a great surface area. The observed maximum current density was of 75 A/m2 . Furthermore, the electrode
preparation is very simple and the design and scaling up could be considered informal but functional, which makes it well-
intentioned for conducting more research. In regard to the metal/metal oxides material, this is not planned to be utilized
directly as anode for MFCs due to corrosion-prone issues. The metals can be extensively employed in electrochemistry
due to good conductivity, extensive potential active sites, and highly catalytic activity (Cai et al., 2020). The metal/metal
oxides modifications greatly enhance the MFCs performance, which significantly decrease the ohmic loss and increases
the bacterial attachment on the anode surface. Alatraktchi et al. (2014) examined the bond between material density and
bacterial colonization by employing Au/CP composite-based anode and demonstrated that it significantly increased in 28%
the efficiency than the observed with a simple CP anode. The improvement was obtained through the utilization of Au
NPs (50–100 nm thickness) on the surface of CP. The higher Au NPs densities improved bacteria community growth on
the surface of the anode, which led to the significant results. Fu et al. (2014) studied the plain graphite (PG) anode coated
with MnO2 /MWCNT and used as anode. The MnO2 /MWCNT addition in PG was conducted through the utilization of the
redox method, which increased in 50% the efficiency of MnO2 /MWCNT/PG anode by minimizing the contact angle and
providing wettability, which ultimately enhanced the kinetic activities. Similarly, MnO2 improved the CP performance by
12
refining its pseudo-capacitive behavior, making available the surface area, and improving the ability of electron transfer
rate. This enhanced the entire working efficiency in 24.7% against the simple CP. However, recently, Ying et al. (2018)
prepared the TiO2 /stainless steel mesh composite anode by employing the electrochemical technique (Fig. 5c and d). This
type of anode presented a higher superiority in biocompatibility toward bacterial growth on the surface of the anode. The
charge transfer resistance was of 3.55 W, which greatly expands the electron transfer rate, the highest achieved current
density was of 70 A/m2 . Zhong et al. (2018) prepared the NiO/PANI composite in petaloid NiO@PANI-CF composite shaped
via in-situ polymerization reaction as shown in Fig. 5e. This composite provided low charge transfer resistance rate, lower
polarization, good biocompatibility, super-hydrophilicity, high surface area and the maximum power density observed in
their experiments was of 1078 mW/m2 . Zeng et al. (2018b) prepared a porous nanostructured Ni0.1Mn0.9O1.45 micro
ellipsoid as good electrocatalyst for the anode oxidation reaction through employing the co-precipitation method as
shown in Fig. 5(f, g). The achieved power density was of 1390 mW/m2 , which might be promoted due to an effective
synergistic effect by enabling the extracellular electron transfer development between the electrogenic active bacteria
and the anode. Guzman et al. (2017) prepared a large surface area and big porosity-based anode by using the nanofiber
mat (polyacrylonitrile (PAN)) through electrospinning. The prepared carbon nanofibers (CNFs) were improved with PEDOT,
which enhanced their conductivities and capacitance. Though, the promising PAN mat have some properties such as non-
conductivity without capacitance. This confirmed that the CNFs anode can be improved via electrodeposition of PEDOT
film. Sonawane et al. (2018a) used electro-polymerization for PANI film coating on stainless-steel (SS) plate with selective
and optimized conditions. High adherence and uniform conductivity of PANI-SS anode were achieved. The observed power
density was of 780 mW/m2 . The discussion showed that the material is required to make the MFCs reaction feasible in
regard to biocompatibility and high energy generation. Earlier, several materials were studied briefly but they all carried
some issues such as cost, material conductivity, a toxic effect toward bacteria etc. The idea of composite is an ideal
approach, particularly in the case of metal oxides with carbonized carbon-based materials, which significantly minimizes
the cost and increases several required properties effectively.
3.2.3. Graphene-based anode material

The Gr‘-based anode material is also considered as modern anode material because Gr‘ derivatives meet the require-
ments to solve the many current challenges such as conductivity, surface area, biocompatibility, and bacterial adhesion.
The natural carbonized carbon conversion into Gr‘ derivatives by using several methods such as hummer method etc.
However, Gr‘ is defined as a 2D crystalline carbon allotrope, which contains the most promising properties e.g., the
observed surface area is 2600 m2 /g, high electric conductivity, which is around 7200 S/m and high mechanical stability
(ElMekawy et al., 2017). The Gr‘-based composite combined with other metal oxides or conducting polymers provides
a breakthrough to increase the application scope of MFCs electrodes due to a high rate of electron movement and high
electrocatalytic activities. It must be considered seriously to upgrade the Gr‘ derivatives through composite with other
materials to apply the MFCs approach in actual world scenarios to meet the current energy supply and wastewater
treatment/pollutant removal crises. Several earlier efforts have shown that Gr‘ modified anode can stimulate bacteria
species to secrete the signaling molecules. These could enhance the bacterial growth and instantaneously assist as
intermediary molecules to increase the efficiency of the electron transfer (Cai et al., 2020). Furthermore, the great surface
area of Gr‘ derivatives offers more active sites and channels for the substrate oxidation and interfacial response, which
promotes the decrease of both, the resistance and activation losses. Sun et al. (2017) studied the Gr‘/PANI hybrid-based
anode and synthesized it by an in-situ method. The Gr‘/PANI hybrid-based anode showed excellent energy yield and
offered higher energy density value than that observed with untreated Gr‘ anode. Other study showed that the Gr/PANI
anode conveyed the synergistic influence on direct electron transfer of S. oneidensis MR-1. Cheng et al. (2018) synthesized
the rGO Au NPs hybrid composite through a layer-by-layer assembly approach. However, this prepared composite coating
was carried out on the surface of CB to fabricate a modified anode. The leaf extract was incorporated to enhance the
hydrophilicity of the composite anode electrode. The achieved energy generation was of 33.7 W/m3 . The high-performance
must be directed toward the good biocompatibility and high rough surface, which is valuable in order to achieve bacterial
colonization. In the meantime, Au NPs increases the electro-catalytic activities and improves the electron transfer rate. On
the other hand, 3D nano-size structure-based anodes also received great attention in MFCs because they can enhance the
material surface area and promote a more active substrate diffusion. Therefore, researchers are devoted to preparing a new
3D Gr‘ anode (Cai et al., 2020). For example, Yang et al. (2016) developed modified GFB (graphite fiber brushes) by using
the graphene oxide (GO) aerogel and later produced 3D-based composite anodes which were quite successful in terms
of operational stability at lab scale. The observed power density was of 2520 mW/cm2 , which is unexpected according
to previous reported work. However, the power bank, so called biofilm would turn out to be denser, this enormously
hindered the substrate diffusion into the internal structure of the anode and therefore, a lower achievement in energy
generation. This consequence suggests that a 3D-based structure is essential for constant generation of electricity in MFCs.
The above-mentioned studies showed that the material improvement can be done by increasing electric conductivity,
surface area and electrocatalytic activities (Cai et al., 2020). The modification of Gr‘ is a future prospect for excellent
performance of MFCs. The Gr‘ modification with metal oxide can serve as excellent anode material such as Gr‘/TiO2 ,
Gr‘/ZnO, Gr‘/CuO and Gr‘/AgO etc. It is also an ideal approach to use natural biomass material as Gr‘-based anode because
this seems to be a cost effective and high-quality material for MFCs. Several Gr‘ modified anode applications in recent
years are shown in Table 3.
13
Table 3
List of Gr‘ modified-based anode, the list presents the recent year progress in regard to MFCs.
Anode material Surface area of the Source of inoculum Substrate Power density Reference
anode (cm2 ) (mW/m2 )
GL/GP 7 Anaerobic sludge Sodium acetate 670 Tang et al. (2015)
GOA/GFB 16 S. oneidensis MR-1 Sodium acetate 2520 Yang et al. (2016)
rGO/MnO 2/ CF 7.1 MFC effluent Sodium acetate 2065 Zhang et al. (2016b)
GO/PANI/GP 2 S. oneidensis MR-1 Sodium acetate 381 Sun et al. (2017)
DGMBE/CF 30 Anaerobic sludge Glucose 112.4 Chen et al. (2017)
GRF/GP 2 S. oneidensis MR-1 Lactate 257 Wang et al. (2017)
Gr/CP 6.7 Anaerobic sludge Simulated wastewater 2381 Yu et al. (2018b)
rGO/Au/CB – Anaerobic sludge Sodium acetate 33.7 Cheng et al. (2018)
GOZ/SSM 36 Anaerobic sludge Simulated wastewater 280 Paul et al. (2018)
rGO/PANI/CC 4.9 Wastewater Sodium acetate 306 Lin et al. (2019)
EL/RGO/CFP 4 E. coli Sodium acetate 1158 Zhou et al. (2019)
rGO-PAM 37.6 Sludge Sodium acetate 758 Chen et al. (2019)
CC–PDA–rGO – Activated anaerobic sludge Acetate 2047 Li et al. (2020)
RGO-CB 131.18 Sewage wastewater – 381 Sayed et al. (2020)
rGO-PDA 120.8 Anaerobic sludge – 988.1 ± 5.2 Liu et al. (2021)
Cellulose derived 71 Synthetic wastewater Sweet potato 1.1 Yaqoob et al. (2021)
GO-PANI
CGP = Crumpled graphene particles; CFP = Carbon fiber paper; GFB = Graphite fiber brush; GOZ = Graphene oxide zeolite; GOA = Graphene
oxide aerogel; GRF = Graphene riboflavin; EL = Eucalyptus leaves; ZnO = Zinc oxide; GO = Graphene oxide; GL = Graphite layer; DGMBE = Dual
graphene modified bioelectrode; CC–PDA–rGO = Carbon cloth-polydopamine-reduced graphene oxide; RGO-CB = Reduced graphene oxide-carbon
brushes; rGO-PAM = Reduced graphene oxide/polyacrylamide; rGO-PDA = Polydopamine decorated reduced oxide graphene; GO-PANI = Graphene
oxide/polyaniline.
4. Bacterial interaction on anode surface
The interaction of the anode surface with the inoculated bacteria is produced by the formation of a biofilm on the
surface of the anode in MFCs. The bacterial biofilm is usually known as powerhouse of MFCs (Sokol and Bradford, 2019;
Yaqoob et al., 2020c). However, the biofilm studies have been focused on several parameters such as dynamic formation,
rate of electron transfer, bacterial growth, and their metabolic activities. It is commonly known that the formation of
biofilm considers multiple and significant phases: such as bacterial attachment on the surface of the anode, growth of
single or multiple layers of bacteria, a polymeric scaffold formation, and development of mature biofilm on the anode. The
bacterial attachment and development of bacterial layers is one of the most essential phases that controls the subsequent
growth of the biofilm (Chuo et al., 2020; Korth et al., 2015). Particularly, the early attachment and single layer of bacterial
adhesion generally depend on the surface properties of the anode material such as hydrophobicity/hydrophilicity, chemical
functional group, and nano-texture. At this point, low electrocatalytic activities and poor energy output are observed at the
anode, due to the poor bacteria densities on its surface. After the maturation stage of the bacterial biofilm, the capability
of reacting and the electrocatalytic activity can be greatly increased (Pinck et al., 2020; Yaqoob et al., 2020d). The chemical
modification is an ideal and active approach to modify the material surface and promote the bacteria attachment along
with an improvement of the electron transfer rate. In wide-ranging, the anode material can be improved by using several
materials in form of composites such as metal NPs, conducting polymers, and chemical or thermal oxidation treatment of
surface. Despite all methods, the electrochemical oxidation is known as the most cost-effective method, in which definite
structural morphologies and functional groups are linked to the anode material surface to promote the healthy bacterial
growth and charge transfer rate (Karthikeyan et al., 2015). Remarkably, the natural waste biomass-based anodes generally
present richness in functional groups. The introduction of the inherent heteroatom has shown a significant variety of
functional groups in the material structure. For instance, Wang et al. (2019) fabricated the pinecone-derived anode through
a simple carbonization process. They observed the good rate of extracellular electron transfer and the potential growth of
exoelectrogens due to the surface modification through N and P dopants. The anode was fabricated through carbonization
at 900 ◦ C and it presents a maximum N content and provides the maximum power density, i.e., 10.88 W/m3 . Similarly,
another important aspect of an anode is the surface roughness, which exerts a direct influence on the formation of anodic
biofilm. Zhang et al. (2014) used bamboo charcoal waste material to fabricate the anode through simple carbonization of
natural bamboo branch wastes. The achieved roughness in the anode surface offers higher availability of active sites and
a high surface area for bacterial attachment as compared to conventional anodes as mentioned above. The authors proved
that the bamboo charcoal-derived anode improved the biofilm formation, enhanced electrogenic bacteria attachment, and
increased the rate of electron transfer from exoelectrogens to the anode over conventional graphite anode. The biofilm
parameters are usually used to improve biofilm development on the surface of the anode and hence advancement in
the mass transfer inside the biofilm of the anode. Therefore, the anode energy response is strictly associated with mass
transport within biofilm and movement of electrons from bacteria to anode. The anode energy response can serve as
external factor to control and stimulate the biofilm maturation. Li et al. (2017) observed the anodic biofilm response
at diverse current densities and noticed that the biofilm formation at a high current density encouraged further active
sites for healthy bacterial attachment, healthier biofilm development and better rate of electron movement than at low
14
Table 4
Comparative profile of biowaste-derived anode and commercial anode electrodes.
Type of material Anode electrode Surface area (cm2 ) Total material cost Power density Cost of power
(US$ ton−1 )a (US$W−1 )b
W/m3 mW/m2
Biomass-derived BCC 32 51–381 4.97 ± 20 457 ± 20 35.79
Biomass-derived BCP 52 51–381 5.78 ± 18 532 ± 18 17.27
Commercial-based GG 0.2 500–800 6.15 ± 5 566 ± 5 392.62
Commercial-based GAC 368 800–2500 7.32 ± 10 674 ± 10 402.80
BCC: biochar derived from forestry waste.

BCP: biochar derived from compressed milling waste.
GGs: graphite granules.
GAC: granular activated carbon.
a
Material costs were obtained from Huggins et al. (2014).
b
Cost of power was obtained by dividing the material cost per reactor by the power density.
current densities. However, a modified anode material can only serve as an ideal anode to upgrade the MFCs approach at
laboratory scale.
5. Economic prospect of natural waste-derived anode electrodes for pilot scale MFCs
The economic feasibility is the most important step for pilot scale MFCs tests. The waste-derived material as anode
can bring significant shifts in costs of the overall MFCs operation. As we already discussed, anode is the most essential
part to carry out the key functions during the operation of MFCs. To date, the non-renewable and unaffordable cost of
the materials are the current challenges. There are very few studies reported on the cost analysis, but some researchers
assessed that the anode material cost could represent around 20% to 50% of the overall MFCs operation cost (Rozendal
et al., 2009; Rabaey et al., 2010; Rodríguez-Couto, 2020). The granular activated carbon (GAC) and graphite granules
(GG) are the most utilized materials in MFCs due to their excellent physiochemical and catalytic properties (He et al.,
2017). The above-mentioned materials imply an inversion of 500–2500 US$ ton−1 , to fabricate the anode electrode. This
is lower than carbon cloth or carbon paper (100,000 to 500,000 US$ m−2 ), which is explained by Huggins et al. (2014).
Despite the lower cost of GAC and GCs than others, they are still not economically feasible to conduct assays at pilot scale.
Additionally, the manufacturing processes of GAC and GCs are a source of pollution, therefore, these materials are known
as poor environment friendly materials (Huggins et al., 2014). However, in the framework of fabricating the cost-effective,
environment friendly anode by using the biomass waste such as waste-derived biochar, or GO derivatives, this seems to be
a promising approach. Thus, many studies have stated the suitability of using biomass waste material to build the anode
electrodes for MFCs. Some examples of these materials are: kenaf (Hibiscus cannabinus) stems, palm kern shell (Offei et al.,
2016), cardboard (Karthikeyan et al., 2015), oil palm waste material (Yaqoob et al., 2020c), sewage sludge and sewage
sludge mixed with fly ash (Jia et al., 2018). Table 4 presents a comparison of the biomass-derived anode (biochar-made
from biowaste) with commercial electrodes in terms of MFCs performance and cost. According to the observations, the
MFCs performance is quite similar but there is a huge difference in cost. The biomass derived anode implied a lower cost
than commercial electrode. Derived from the extensive literature review, it was found that there is very little information
reported regarding the cost comparison between the biomass-derived and commercial anodes. In order to advance in this
field of research, further studies on the technical and economic feasibility of waste-derived anode for MFCs are required.
6. Problems and future perspective of anode material
In the previous decades, several efforts have been done in the development of anode materials to enhance the overall
performance of MFCs. Despite all the developments, the existing model of MFCs is not yet a suitable technique at large
scale application. There are several problems that need to be addressed in anode development, which is a major hot spot
of research to apply MFCs at industrial level in the future. The present challenges and problems in the development of
anode are cost of the anode material because anode is playing a vital role in MFCs along with other factors. Therefore,
the cost of the anode material makes the entire MFCs approach economically unsuitable for commercial scale. Several
conventional materials have been studied in the last decade to minimize the cost, but conventional materials are not
capable of producing sufficient energy and pollutant removal to meet the current global demands. Along the same
lines, another problem is the design and configuration of the anode electrode because a proper design and scaling of
the anode are key factors to provide definite surface area for bacterial attachment and for further required actions
(Asim et al., 2021). Therefore, the selection of the anode material is also a critical stage to fabricate an ideal anode
for better MFCs performance. An improper design and/or configuration disturbs the growth of healthy bacteria and
offers less opportunities to easy the electron flow efficiently. The long-term stability is also one of the most important
challenges for modern research because all the conventional materials have failed to serve for long term and continuous
MFCs operation. Durability of the anode is highly questionable, as it cannot sustain the stability for longer periods of
15
time due to poor mechanical and chemical stability (Hindatu et al., 2017). Consequently, the future research in the
area of anodes development for practical application in MFCs must be focused on the following aspects: The natural
biomass waste materials as the most promising source to develop a high-quality anode electrode at suitable cost. As it
was summarized in Table 1 in regard to the several biomass waste-derived anodes along with their significant results.
However, the 3D macropore-based biomass anode such as loofah, pomelo peels and kenaf have shown a superior energy
production. Furthermore, the natural resources derived anodes are cost-effective and easily available, which can fulfill
the basic requirement of an ideal anode: easily available at reasonable cost (Cai et al., 2020). Several materials can offer
high performance after minor modifications, but their availability and cost make them unsuitable, such as in the case
of the metal-based materials. The natural biomass waste materials also offer rational surface area with adjustable pore
size, this is important because pore size exerts a major effect on performance, the larger the pore size of the anode
material the larger the disturbance in the extracellular electron transfer mechanism. The carbonization process provides
an opportunity to adjust the pore size of the material according to the requirement and natural biomass-derived material
can be further modified by using other materials to enhance its electrocatalytic activity and electrical conductivity. At
present time, a 3D printing method is moderately developed to maintain the anode architecture. The proper designing
and scaling can be carried out by using 3D printing technology to increase the surface area and tune the definite pore
size (Cai et al., 2020). Thus, it is a good approach to improve the bacterial attachment and the electrocatalytic sites
without blocking the substrate diffusion. Furthermore, to address the long-term stability issue of modern anode, the
modification strategies need to be adapted because multidisciplinary research may spark surprising outcomes. Natural
biomass resources, metal/metal oxides, conducting polymers, CNTs and commercial Gr‘ have shown several advantages
and disadvantages as is presented in Table S3. Therefore, the idea of composite material is a superlative approach to
minimize the material disadvantages by improving their positive aspects further. The natural waste-derived materials
can be further treated to achieve Gr‘-based materials, which are also considered as modern materials to obtain efficient
results from MFCs. The modification of natural biomass-derived Gr‘ derivatives with metal oxides can offer unexpected
results as is the case of GO/ZnO, GO/TiO2 , GO/CuO, GO/AgO etc. The natural biomass-derived Gr‘ derivatives usually showed
low mechanical strength and high electric resistance, but metal oxides showed high mechanical strength and low electric
resistance. Therefore, the composite of such materials may trigger the significant outcomes. As it stands now, substantial
work is still required to apply the MFCs at large scale.
7. Conclusions
This review article extensively illustrates the role of the anode material in MFCs along with a discussion on the several
types of anode materials. The review clearly demonstrates that the anode has an essential role in MFCs implemented
for wastewater treatment and energy generation, which usually involves the oxidation of an organic substrate in the
anode. In previous decades, many conventional anode materials were introduced, but they have failed to meet the modern
demands. An ideal anode needs to present some basic properties that lead to the obtainment of significant results.
Among all the materials, the natural resources and their composite are good materials to fulfill the modern requirements
while minimizing other disadvantages. The natural biomass-derived materials exhibited excellent bacterial attachment
on surface as mentioned in Section 4 because the bacterial growth, their attachment and respiration processes are also a
key factor in MFCs operation. Therefore, for an appropriate MFCs operation, the anode should be both, technically feasible
and flexible. The natural biomass derived material composites are necessary to minimize some drawbacks such as the low
mechanical power and high electrical resistance. These problems should be addressed by using metal oxide composites
because they can offer high mechanical strength and minimize the electrical resistance. Finally, a more cost-effective and
high-performance MFCs technique is preferable in modern world to meet the present energetic and water pollution crises.
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have
appeared to influence the work reported in this paper.
Acknowledgments
This review article was supported by Universiti Sains Malaysia, (Malaysia).
Appendix A. Supplementary data
Supplementary material related to this article can be found online at https://doi.org/10.1016/j.eti.2021.101579.

16
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Environmental Technology & Innovation 23 (2021) 101579

Contents lists available at ScienceDirect

Environmental Technology & Innovation

Modern trend of anodes in microbial fuel cells (MFCs): An

2. MFCs: Mechanisms of energy generation and pollutant remediation

2.1. Mechanism of electron transfer from bacteria to anode electrode

2.2. Reduction mechanism of pollutant (Heavy metals)

Hg2 Cl2 (s) + 2e− −→ 2Hg(l) + 2Cl−

2Cu2+ + H2 O + 2e− −→ Cu2 O + 2H+

2.3. Biochemical cell reactions and electrochemical measurements

(a) If glucose is used as organic substrate

3.1. Conventional anode materials

3.2. Modern anode material

3.2.1. Natural biomass-based anode material

3.2.2. Composite-based anode material

(continued on next page)

3.2.3. Graphene-based anode material

4. Bacterial interaction on anode surface

BCC: biochar derived from forestry waste.

6. Problems and future perspective of anode material

Declaration of competing interest

This review article was supported by Universiti Sains Malaysia, (Malaysia).

Appendix A. Supplementary data

Supplementary material related to this article can be found online at https://doi.org/10.1016/j.eti.2021.101579.

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