processes

Article
Impact of Self-Fabricated Graphene–Metal Oxide Composite
Anodes on Metal Degradation and Energy Generation via
a Microbial Fuel Cell
Akil Ahmad 1, * , Mohammed B. Alshammari 1 and Mohamad Nasir Mohamad Ibrahim 2

1 Department of Chemistry, College of Science and Humanities in Al-Kharj, Prince Sattam bin Abdulaziz
University, Al-Kharj 11942, Saudi Arabia
2 Materials Technology Research Group (MaTRec), School of Chemical Sciences, Universiti Sains Malaysia,
Minden, Penang 11800, Malaysia
* Correspondence: [email protected]

Abstract: Microbial fuel cells (MFCs) are thought to be ecologically friendly, despite electron transport
and generation challenges. In order to address this, the efficiency of MFCs was investigated using
two different anode electrodes made from biomass: graphene oxide (GO) and graphene oxide-metal
oxide (GO-MO) (GO-ZnO). After 18 days of operation, the maximum power density for GO was
0.69 mW/m2 , whereas the maximum power density for GO-ZnO was 1.05 mW/m2 . Furthermore,
the ability of MFCs to transform the soluble metal ions (Cd2+ , Cr3+ , Pb2+ , and Ni2+ ) into an insoluble
state was investigated, which is a secondary use of MFCs with significant benefits. In the soluble
state of metal ion transformation into an insoluble state, the rate of GO-ZnO was higher (92.71%)
than that of GO (81.20%). The outcomes of material, analytical, and biological tests undertaken to
validate the efficiency of anodes are presented. It has been shown that using innovative materials
as electrodes in MFCs is a potential method for improving electron transport. Furthermore, as an
organic substrate, food waste seems to be a viable alternative to more traditional options. In light
of these discoveries, we investigate various unanswered issues and possibilities for MFCs. Organic
Citation: Ahmad, A.; Alshammari, substrate evaluation trials were also included in the present results to demonstrate that organic waste
M.B.; Ibrahim, M.N.M. Impact of may be a reliable source of MFC performance. This article also has a thorough discussion of food
Self-Fabricated Graphene–Metal
waste oxidation, as well as challenges and future recommendations.
Oxide Composite Anodes on Metal
Degradation and Energy Generation
Keywords: energy transportation; microbial fuel cell; biomass; electrodes; wastewater treatment
via a Microbial Fuel Cell. Processes
2023, 11, 163. https://doi.org/
10.3390/pr11010163

Academic Editors: Georgios Bampos, 1. Introduction

Georgia Antonopoulou, Zacharias
A microbial fuel cell, also known as an MFC, is a developed type of bio-electrochemical
Frontistis and Qunjie Xu
system that generates electrical power by efficiently oxidizing organic substrates [1]. The
Received: 29 November 2022 bacterial species that have been introduced act as a catalyst, which speeds up the process
Revised: 1 January 2023 of oxidizing the organic substrate [2]. An MFC may be used in a wide variety of contexts,
Accepted: 3 January 2023 and research on MFCs has become more popular in recent years [3]. Modern society faces
Published: 5 January 2023 wastewater treatment and energy problems. Thus, an MFC is becoming a popular method
in the present era. Recent research has shown that metal ion degradation via MFCs is the
most successful because electron acceptors can be used to boost energy production [4,5]. It
is better to use MFCs for efficient metal ion recovery or remediation.
Copyright: © 2023 by the authors.
According to the literature, low energy performance makes the MFC unsuitable for
Licensee MDPI, Basel, Switzerland.
industrial use. The low energy performance emphasized electron generation and electron
This article is an open access article
transportation from anode to cathode electrodes. MFCs perform poorly due to insuffi-
distributed under the terms and
conditions of the Creative Commons
cient electron transport from bacteria to electrodes [6]. The transportation of electrons is
Attribution (CC BY) license (https://
a major challenge. Anode electrodes directly influence electron production and transfer
creativecommons.org/licenses/by/
capability [7]. To perfect this process, the problem must be addressed by meeting the
4.0/).

Processes 2023, 11, 163. https://doi.org/10.3390/pr11010163 https://www.mdpi.com/journal/processes

Processes 2023, 11, 163 2 of 19

aforementioned requirements [8]. Typically, carbon and pure metal materials were em-
ployed as electrodes, but during operation, the metal-based anodes demonstrated corrosion
problems that harmed long-term stability as well as the development of bacteria [9]. The
mild conductivity, large surface area, and easy availability of carbon materials led to their
selection. The performance of MFCs derived from traditional carbon-based materials was
insufficient [10].
According to a previous literature survey, it was proposed that graphene derivatives
are the potential source for anode development [11,12]. The surfaces of waste-derived GO
materials are strongly oxygenated, which may drastically modify Van der Waals interac-
tions and result in a variety of water solubilities [13]. The edge plane nanobands on the
heterogeneous surfaces of GO are primarily electroactive, while the basal plane islands
are electrochemically inert [14]. The outstanding conductivity, surface area, biocompati-
bility, and mechanical stability of graphene derivatives were revealed. As a result, when
compared to other materials, they may be regarded as great performers [15]. Graphene
derivatives also have several flaws, including high manufacturing costs and a lack of
resources. One solution to this problem is to synthesize graphene oxide (GO) from forest
waste, notably oil palm material. This method is both affordable and readily accessible.
It was previously reported that the GO made from forest debris was biocompatible, had
a large surface area, and was an excellent conducting material [16]. Additionally, the
GO generated from forest waste must be modified with metal oxide to boost the electron
transfer efficiency (ETE), as this will increase its conductivity. It is preferred that the metal
oxide be readily accessible, inexpensive, highly conductive, biocompatible, and exhibit
excellent ETE. According to several studies, metal oxides were added to electrodes as
modifiers to improve the performance [17,18]. Forest biomass-derived GO was enhanced
with environmentally-friendly synthetic commercial zinc oxide (ZnO) nanoparticles (NPs)
to increase the electron transport rate. According to previous evidence, ZnO was thought
to be the most promising of all the metal oxides because of its exceptional electrical, semi-
conductor, and optical activities as well as its high electrocatalytic ability [19]. For example,
Shakeel et al. [20] investigated the ZnO-coated polyindole-functionalized multiwalled
CNTs and their use as a biofuel cell anode. The resulting output was high, but the biofilm
was damaged due to the CNTs’ cellular toxicity.
Another issue is the generation of electrons since there is not enough organic substrate
available. Energy efficiency and metal reduction directly rely on the rate of electron
production, making it amongthe key challenges [21]. While the production rate was
determined to be low, some investigations suggested that carbon derivatives successfully
transfer electrons from anode to cathode [22]. The high rate of electron production is
inhibited by the bacterial species’ limited access to organic substrate. Recent research by
Fadzli et al. [21] shown that using highly carbohydrate-based organic substances is essential
for enhancing energy efficiency. Additionally, they stated that using organic substrates
obtained from waste, such as waste food, fruit waste, and sugar waste, is the best approach.
Organic matter, which is generated by a variety of sources, includes food waste. Annual
increases in food waste were observed [23]. As waste management grows more challenging,
food waste is seen as a major concern. Thus, several studies supported more efficient food
waste treatment and its advantages. Food waste as an organic substrate in MFCs is used
in this study. Food waste serves as a source to generate energy and degrade the mixture
of Cd2+ , Cr3+ , Pb2+ , and Ni2+ ions in synthetic wastewater via MFCs. This research also
provides new research for other researchers by investigating the effect of waste organic
substrate with self-fabricated electrodes on MFC performance.

2. Methodology Details
2.1. Material and Reagent
Oil palm biomass waste (received a gift from Malaysia), sulfuric acid (97%, Sigma-
Aldrich, Berlin, Germany), potassium permanganate (Sigma-Aldrich), hydrogen peroxide
(30–32%,—Sigma-Aldrich), hydrochloric acid (37–38%,—Sigma-Aldrich), ethanol (95%,
Processes 2023, 11, 163 3 of 19

QRec), zinc oxide (Sigma-Aldrich), polysulfones (PSF, Sigma-Aldrich), lead, cadmium,

chromium, nickel nitrate (Sigma-Aldrich), dichloromethane (QRec), sodium nitrate (Sigma-
Aldrich), commercial graphite rod (FUDA Lead, New York, NY, USA), glucose (AR grade)
and distilled water (DW) were used in this research.

2.2. Preparation and Synthesis of Oil Palm Biomass-Derived GO

Carbon powder was produced by carbonizing the oil palm biomass. The dried
oil palm biomass goes through carbonization by being heated up (800 ◦ C) at a rate of
20 ◦ C per minute while being continuously surrounded by argon gas for 3 h. The resulting
carbonized flakes were put through a mortar and processed quickly to produce powder.
The produced oil palm biomass-carbon is then subjected to the Hummer method to produce
GO [24]. The oil palm biomass-carbon (10 g) and NaNO3 (15 g) were mixed in 300 mL
of H2 SO4 with constant stirring. The mixture was then placed in an ice bath (0–5 ◦ C). At
this temperature, the mixture was agitated for two hours before KMnO4 (25 g) was gently
added to the suspension. After the stirring was finished, the mixture was taken out of the
ice bath and left to stir continuously for two days so that the oxidation process could be
completed at room temperature. When the color became violet, brown, DW (140 mL) was
gradually added and stirred once more at 85 to 90 ◦ C until a dark brown color was visible.
By swirling continuously while 210 mL of DW was added, this solution was further diluted.
A total of 15 mL of H2 O2 is then added to the solution to cease the reaction. The stored
mixture was further cleaned with ethanol and water by centrifugation, before being dried
at 45 ◦ C to produce the GO.

2.3. GO-ZnO Composites Preparation via Solvothermal Method

The solvothermal approach was used to produce the GO-ZnO composites. To make
the GO-ZnO composite, 250 mL of ethanol and 5 g of GO along with 2.5 g of commercial
ZnO were mixed at room temperature and agitated for 3 to 4 h. The particle size of the ZnO
was less than 120 nm according to the manufacturing company. At 210 ◦ C, the finished
product was heated until it became powder (GO-ZnO composite).

2.4. Material Characterizations

Multiple approaches were used to characterize the synthesized materials (GO and
GO/ZnO composite). The functional groups and morphology of the synthesized material
were investigated using the Fourier Transform Infrared (FTIR; Perkin Elmer model System
2000) and scanning electron microscope (SEM) (SEM-Zeiss, model DSM-960, Germany). The
atomic force microscopy (AFM-SII Sciko Instrument INC SPI 3800N Probe station), which
was also used to assess the material’s surface roughness. Most often, in material science,
the X-ray diffraction (XRD; Philips PW 1710 X-ray diffractometer) method is employed to
determine the crystalline structure of produced material. The primary purpose of the XRD
is to determine a material’s structure from its diffraction pattern.

2.5. Fabrications of Self-Fabricated Anodes

For the fabrication of two different kinds of electrodes in MFC operations, the prepared
materials were used. The performances of two electrodes were investigated using various
electrochemical methods. A 2.2 mm graphite rod served as the substrate and current
collector for each experiment while fabricating anodes. To produce the GO anode, PSF
solution (3.5 g solution in 10 mL dichloromethane), 5 g of GO powder was thoroughly
mixed. The graphite rod was wrapped in the resulting paste-like substance, which was
then heated at 45 ◦ C for two to three days to reinforce it mechanically. Similar to this,
to produce the paste-like materials, 5 g of GO-ZnO composites was mixed individually
with 3.5 g PSF solution. To produce the rod-shaped anodes, the paste-like ingredients
were applied all around the graphite rods. Next, both electrodes were heated at 400 ◦ C for
24 h. Again, both electrodes were briefly immersed in PSF solution prior to MFC operations.
The PSF serves as a supporting and protecting material in electrode fabrication. The PSF
 then heated at 45 °C for two to three days to reinforce it mechanically. Similar to this, to
produce the paste-like materials, 5 g of GO-ZnO composites was mixed individually with
3.5 g PSF solution. To produce the rod-shaped anodes, the paste-like ingredients were
applied all around the graphite rods. Next, both electrodes were heated at 400 °C for 24 h.
Processes 2023, 11, 163 Again, both electrodes were briefly immersed in PSF solution prior to MFC operations. 4 of 19
The PSF serves as a supporting and protecting material in electrode fabrication. The PSF
not only helps to bind the synthesized material, but it also reduces the Zn leakage during
the
not MFC operation
only helps [25].the
to bind Allsynthesized
the prepared anodesbut
material, were 8 cmreduces
it also in length,
the1Zn
cmleakage
in radius, and
during
had 56.52 cm 2 of surface area. The biomass conversion to GO-ZnO composite for anode
the MFC operation [25]. All the prepared anodes were 8 cm in length, 1 cm in radius, and
electrode
had 56.52fabrication is shown
cm2 of surface area.in Scheme
The 1. conversion to GO-ZnO composite for anode
biomass
electrode fabrication is shown in Scheme 1.

Scheme 1. The conversion of oil palm biomass to GO results in the formation of a GO-ZnO composite
Scheme 1. The conversion
for the fabrication of anodeofelectrodes
oil palm biomass to GO results
for single-chamber in the formation of a GO-ZnO com-
MFCs.
posite for the fabrication of anode electrodes for single-chamber MFCs.
2.6. Organic Substrate, Inoculation, Assembly and Operation of Microbial Fuel Cell
2.6. Organic
The foodSubstrate, Inoculation,
waste was collectedAssembly
from theand Operation
local of Microbial
restaurants. For theFuel Cell
experiment, 500 g of
foodThe
waste
food waste was collected from the local restaurants. For the experiment, 500were
was used. Fish, rice, sweet cake, and curry were among the foods that g of
thrown
food away
waste was asused.
food Fish,
waste. Thesweet
rice, foodcake,
wasteandwascurry
separated from the
were among theplastic
foods and
thatbones
were
before being
thrown away processed for 5 The
as food waste. min food
in a blender.
waste was Forseparated
the electrical
frompulverizer
the plastictoand
grind the
bones
food waste to perfection, distilled water (50 mL) was supplied. To minimize
before being processed for 5 min in a blender. For the electrical pulverizer to grind the clogging
issues,
food the food
waste waste wasdistilled
to perfection, subsequently
water filtered
(50 mL)using a stainless-steel
was supplied. sieve toclogging
To minimize remove theis-
coarse elements. The concept of gravity helped to remove any oil that was
sues, the food waste was subsequently filtered using a stainless-steel sieve to remove floating on the
the
surfaceelements.
coarse of the filtered liquid. The
The concept wastewater
of gravity helpedwas collectedany
to remove from
oil the
thatlocal
was pond,
floatingand
onthen
the
a concentration of 10 mg/L of Cd2+ , Cr3+ , Pb2+ , and Ni2+ was added to the wastewater.
surface of the filtered liquid. The wastewater was collected from the local pond, and then
On the basis of suggestions from the literature, a supplement of 10 mg/L was added to
a concentration of 10 mg/L of Cd2+, Cr3+, Pb2+, and Ni2+ was added to the wastewater. On
the effluent [26]. In this research work, the wastewater that has metal ions added to it is
the basis of suggestions from the literature, a supplement of 10 mg/L was added to the
referred to as “synthetic wastewater”. Table 1 presents a comparison of the characteristics
effluent [26]. In this research work, the wastewater that has metal ions added to it is re-
of local pond water with those of synthetic wastewater.
ferred to as “synthetic wastewater”. Table 1 presents a comparison of the characteristics
of local pond water with those of synthetic wastewater.
Table 1. Recorded characteristics of local pond and synthetic wastewater.

Parameters Local Pond Wastewater Synthetic Wastewater

pH 6.97 6.40
Color Light yellow Light black yellowish
Temperature 24 ± 1 ◦C 24 ± 1 ◦ C
Metal ions O mg/L 10 mg/L
Odor Bad smell Bad smell
Processes 2023, 11, 163 5 of 19

In this study, an MFC with a single chamber was used. A single-chamber MFC has
the following components: a container with a capacity of 1000 mL for water; graphite rods
serving as the cathode; GO, GO-ZnO serving as the anode; resistance; and crocodile clips.
The graphite rod’s length was measured at 9.5 cm, and its width was determined to be
1 cm. The anode-cathode were split by a distance of 4.5 cm. The chamber of the MFC had a
total weight of 1 kg worth of food waste put into it. In addition to the food waste, there was
a total of 700 mL of synthetic wastewater. The anode was immersed into the food waste,
which served two purposes: it was the location where the bacteria attached themselves,
and it was also the location where electrons were generated. However, the cathode rod was
placed into the topmost layer of wastewater. The graphite rods were outfitted with crocodile
clips to secure various items. The external resistance was 1000 Ω. Internal and external
resistance should normally be equivalent. Due to the formation of the biofilm surrounding
the anode at the beginning of the operation, the internal resistance steadily decreased. As
a result, using a certain external resistance is not a strict rule. Anode to cathode distance
and surface area are two factors that often affect a cell’s internal resistance [27]. The
1000 external resistor is preferred for MFC operation while operating at room temperature
and a pH of 6 to 7, according to a prior research on the influence of external resistance
that was published by Igboamalu et al. [28]. Additionally, 1000 was used prior to the
operation for a few days to validate the accurate selection of the external resistance, and
it was noted that the closed-circuit voltage was lower than the open circuit voltage when
loaded by the external resistance, but that it later slowly recovered. In general, larger
resistance should be used if it cannot be recovered, and lower resistors should be used
in open circuit voltage if there was no appreciable voltage drop [29]. In the Microbiology
Teaching Laboratory, the temperature was allowed to remain at room temperature. A mix
of cultural bacterial species were present in the wastewater. To always keep the appropriate
amount of oxygen in the reactor, an air pump was put there. During this technique,
2 mL of a glucose solution (15 g in 250 mL of liquid) was infused daily to make up for the
water that was lost as a result of evaporation. The glucose has a dual use: it replaces the
water that was lost, and it also provides the nutrients that are necessary to stimulate the
activity of the bacteria. The glucose also functions as an electron donor when the substance
is subjected to electrochemical oxidation. It is possible that the MFC’s performance will
improve significantly as a result of this modification. The whole experiment took place in a
continuous trend at room temperature for a period of 30 days. In addition, the experiment
was carried out not once, not twice, but three times to guarantee that the outcomes would
be the same each time.

2.7. Analytical Measurements

To evaluate the anodes’ efficiency in energy production, electrochemical characteriza-
tions were carried out. Various external resistance values (100 to 5000 Ω) were connected
to monitor the voltage production trend or polarization behavior. A digital multimeter was
employed to record the measurements, and the Ohm law was then used to compute the cur-
rent density (C.D.) and power density (P.D.) The P.D, C.D. and current calculation formulas
are provided elsewhere [30]. In each experiment, the first measurement was made using a
new supply of inoculum with a constant concentration. Each resistance value fluctuation
needed 30 min to settle before the next resistance measurement could be taken. To examine
the redox potential of the cells, a cyclic voltammogram (CV) using a three-electrode setup
was used. The electrodes were composed of Pt wire, glassy carbon electrodes, and reference
electrodes made of Ag/AgCl (counter electrode). Up until the reaction was finished, the
CV analysis was carried out every 10 days. Within the potential range of +0.8 to 0.8 V, the
pace of the CV scan was 10 mV/s. The CV peaks at various time intervals were further
analyzed using specific capacitance (Cp ) to assess the bioelectrochemical performances
of the bacterial population during MFC operation. Cp is often defined as the integration
Processes 2023, 11, 163 6 of 19

across the whole set of statistics per unit area of both the cathode and the anode. Cp is
measured in F/g. To determine Cp , the following Equation (1) was used.

A
Cp = (1)
2mk(∆V )

where A = CV area, m = load, k = scan speed, and V is the whole voltage range of the
CV analysis.

2.8. Biological Characterizations and Parameter Optimizations

The degradation of metal ions was measured using a PerkinElmer AAnalyst
400 atomic absorption spectrometer after 5 days, as per the anticipated timetable, using a
sample of 2 mL liquid taken from the MFC. From the findings of the AAS, the degradation
efficiency rate was evaluated using Equation (2).

Tintial − Tfinal
RE = × 100 (2)
Tintial

where RE = degradation efficiency, Tfinal = final concentration and Tinitial = initial concentration.
SEM evaluates bacterial species’ compatibility with anode electrodes. SEM images
revealed the presence of bacteria on the anode. Only organic substrate factors, using GO
and GO-ZnO as anodes, were the focus of our research as we examined the direct influence
these parameters had on the performance of MFCs. In general, it is necessary to tune the
settings for GO and GO-ZnO anodes since these factors directly impact electron transport
and the development of bacteria. To get the most accurate results possible for both the
GO and the GO-ZnO, several experimental runs were carried out. Other factors, such as
external load, temperature, and electrodes, were held constant throughout the experiment.
In this study, commercial glucose and food waste were used as comparisons to demonstrate
that food waste is superior as an effective organic source of sugar. After ten days, measures
of energy consumption and the efficiency of deterioration were recorded. Throughout
the process of optimizing the organic substrate, we continued to use the same inoculation
source for operation.

3. Results and Discussion

3.1. Synthesized Material Analysis
The investigation of FTIR spectroscopy shows the GO and GO-ZnO formation in
Figure 1. FTIR spectroscopy clearly showed that material oxygen functional groups were
representative GO’s. These functional groups were ascribed to C–O–H deformation, C–
O vibrations, C–OH stretching vibrations, and the C=O stretching of COOH groups by
peaks at 1230, 1090, 1400, and 1700 cm−1 . The 1050 cm−1 band represented C-O stretching
vibrations combined with C–OH bending. Bands near 1690 cm−1 , which were related
to the C=C skeletal vibrations of GO, were also observed with ease. In addition to this,
a wide peak appeared at approximately 3400 cm−1 , and it was thought that this peak
corresponded to the C–OH groups; hydroxyl stretching vibrations exist [31]. These findings
are comparable with those that have been reported in the literature, which have demon-
strated that the existing oxygen groups significantly increase the hydrophilic properties of
GO as well as the interlayer spacing. These findings are supported by these results. The
presence of another strong peak at approximately 560 cm−1 , which can be attributed to
the characteristic stretching vibration modes of Zn-O and which confirmed the attachment
of ZnO nanoparticles to GO following solvothermal treatment, was evidence that ZnO
nanoparticles had been successfully attached to GO [32].
 another strong peak at approximately 560 cm−1, which can be attributed to the character-
istic stretching vibration modes of Zn-O and which confirmed the attachment of ZnO na-
Processes 2023, 11, 163 7 of 19
noparticles to GO following solvothermal treatment, was evidence that ZnO nanoparticles
had been successfully attached to GO [32].

Figure 1. FTIR
Figure 1. FTIR spectra
spectra of
of the
the synthesized
synthesized material.
material.

In
In addition
addition to
to this,
this, the
the crystalline
crystalline nature
nature ofof the
the synthesized
synthesized composite,
composite, which
which waswas
shown
shown by XRD examination as presented in Figure 2, provided◦further evidence that
by XRD examination as presented in Figure 2, provided further evidence that GO
GO
was
wassynthesized.
synthesized.TheThereason
reasonfor forthis
thiswas
wasaastrong
strongpeak
peakat at2θ
2θ== 10
10°.. This
This peak
peak was
was precisely
precisely
matched ◦
matched with
with GO’s
GO’s (001)
(001) reflection.
reflection. AA diffraction
diffraction peak
peak at at 2θ
2θ== 24.2
24.2° showed
showed successful
successful
◦ indicated the
graphite oxidation [33]. According to the literature, the strong peak at 2θ = 10
graphite oxidation [33]. According to the literature, the strong peak at 2θ = 10° indicated
successful synthesis of the GO material [33–35]. At the same time, the GO-ZnO XRD model
the successful synthesis of the GO material [33–35]. At the same time, the GO-ZnO XRD
was compatible with the hexagonal wurtzite ZnO structure [36]. The most prominent
model was compatible with the hexagonal wurtzite ZnO structure [36]. The most promi-
peaks were found at 2θ values of 33.70◦ , 36.101◦ , 38.141◦ , 47.02◦ , 56.602◦ , 59.761◦ , and
nent peaks were found at 2θ values of 33.70°, 36.101°, 38.141°, 47.02°, 56.602°, 59.761°, and
68.02◦ , respectively. These values corresponded to the lattice planes (001), (100), (002),
Processes 2022, 10, x FOR PEER REVIEW 8 of(101),
20
68.02°, respectively. These values corresponded to the lattice planes (001), (100), (002),
(102), (110), and (112); additionally, the XRD results showed that the ZnO particles were
(101), (102), (110), and (112); additionally, the XRD results showed that the ZnO particles
successfully embedded on GO [37].
were successfully embedded on GO [37].

XRD
Figure2.2.XRD
Figure spectraofof(i)(i)GO
spectra GOand,
and,(ii)
(ii)GO-ZnO
GO-ZnOcomposite.
composite.

Ascan
As canbebeseen
seenininFigure
Figure3,3,SEM
SEMwas
wasapplied
appliedtotoinvestigate
investigatethe
thesurface
surfacemorphology
morphology
of both graphene oxide (GO) and the GO-ZnO composite. SEM pictures demonstrate
of both graphene oxide (GO) and the GO-ZnO composite. SEM pictures demonstrate the the
simple form of the GO that may have been exfoliated as well as the particle size range
between 100 and 200 nm. The SEM scan reveals that the average size of the ZnO was less
than 120 nm. In addition, a disk-like structure of ZnO NPs in the form of a hexagon was
seen in the SEM picture. This structure was surrounded by six uneven edges. The review
 Figure 2. XRD spectra of (i) GO and, (ii) GO-ZnO composite.
Processes 2023, 11, 163 8 of 19

As can be seen in Figure 3, SEM was applied to investigate the surface morphology
of both graphene oxide (GO) and the GO-ZnO composite. SEM pictures demonstrate the
simple
simple form
form ofof the
theGOGOthat
thatmay
mayhave
havebeen
beenexfoliated
exfoliated as
aswell
wellasasthe
theparticle
particlesize
sizerange
range
between
between100100and
and200 200nm.
nm.The
TheSEM
SEMscan
scanreveals
revealsthat
thatthe
theaverage
averagesize
sizeof
ofthe
theZnOZnOwas
wasless
less
than 120 nm. In addition, a disk-like structure of ZnO NPs in the form of
than 120 nm. In addition, a disk-like structure of ZnO NPs in the form of a hexagon was a hexagon was
seen
seenin
inthe
theSEM
SEMpicture.
picture.This
Thisstructure
structurewaswassurrounded
surroundedby bysix
sixuneven
unevenedges.
edges.TheThereview
review
of
of the relevant prior literature led to this conclusion [38]. The ZnO particles presenceon
the relevant prior literature led to this conclusion [38]. The ZnO particles presence on
the
theGO
GOsurface
surfacewaswasshown
shownby bythe
theGO-ZnO
GO-ZnOcomposite.
composite.The Thecomposite
compositesurface
surfaceshowed
showedthethe
presence
presenceofofhexagonal
hexagonalcrystal
crystalZnO
ZnOparticles,
particles,along
alongwith
withaaparticle
particlesize
sizeof
of less
less than
than 200
200 nm.
nm.
The
The SEM from this investigation were compared to previously published data, andthe
SEM from this investigation were compared to previously published data, and the
results
resultswere
wereinterpreted
interpretedappropriately
appropriately[38].
[38].

Figure 3. SEM analysis of (i) GO and, (ii) GO-ZnO composite.

Processes 2022, 10, x FOR PEER REVIEW
Figure 3. SEM analysis of (i) GO and, (ii) GO-ZnO composite. 9 of 20

Figure44presents
Figure presentsthe
theresults
resultsof
ofananinvestigation
investigationperformed
performedwith withan
anAFM
AFMon onGO
GOand
and
GO-ZnO.
GO-ZnO. The picture obtained
The pictureofobtained by
by AFMAFM of GO revealed a few layers of carbon with an
uneven composition GO particles with of GO revealed
a smooth a The
surface. few GO-ZnO
layers of composite
carbon with AFMan
uneven composition
picture showed, of GO
without particles
a doubt, thewith a smooth
influence thatsurface.
the ZnO The GO-ZnO
particles had composite AFM
on the surface
picture showed, without a doubt, the influence that the ZnO particles had on the
of the GO sheet. This indicates that the GO in the composite exhibited positive exfoliation surface of
the GO sheet. This indicates that the GO in the composite exhibited positive
behavior. According to the research that was performed in the past, the surface roughness exfoliation
behavior. According
that was shown to GO-ZnO
by the the research that wasmay
composite performed
serve asinconfirmation
the past, thethat
surface roughness
the material is
that was shown by the GO-ZnO composite may serve as confirmation that the material is
an appropriate location for the development of bacteria [39].
an appropriate location for the development of bacteria [39].

Figure 4. AFM
Figure 4. AFM image
image of
of (i)
(i) GO
GO and,
and, (ii)
(ii) GO-ZnO
GO-ZnO composite.
composite.

3.2. Energy Generation Analytical Measurement Studies

Experiments using the GO and GO-ZnO composites were carried out in MFCs for
the purpose of analyzing the trends in voltage production at the anodes. Each experiment
lasted for a period of 30 days. During the first few days of operation, a considerable vari-
ation in the voltage generation was seen, with GO producing 1 mV and the GO-ZnO com-
Processes 2023, 11, 163 9 of 19

3.2. Energy Generation Analytical Measurement Studies

Experiments using the GO and GO-ZnO composites were carried out in MFCs for
the purpose of analyzing the trends in voltage production at the anodes. Each experiment
lasted for a period of 30 days. During the first few days of operation, a considerable
variation in the voltage generation was seen, with GO producing 1 mV and the GO-ZnO
composite anode delivering 2 mV, respectively. The voltage that was produced by GO was
111 mV, whereas the voltage that was generated by the GO-ZnO composite was 137 mV
on day 18. The next day, the GO-ZnO composite produced 137 mV, which was more than
GO’s previous output of 111 mV. In both instances, there was a consistent upward trend in
voltage; however, the GO-ZnO composite produced a far higher voltage than GO. In the
beginning, the bacterial colony slowly oxidized the organic substrate that was fed, and as a
result, the voltage steadily grew. According to the findings of the GO-ZnO composite, the
addition of ZnO improved electron transport and made the material biocompatible since it
did not have any adverse effects on living bacteria. Up until the point when the voltage
production stopped, both electrodes were functioning normally. After 18 days, there was a
clear decreasing trend in the voltage generation of both electrodes, and later it was very low.
The initial fresh growth of bacterial species in the anode chamber caused this continuous
rise when the bacteria quickly began oxidation and electron transport was strong. During
the process of oxidation of the organic substrate, the newly constructed anode made it
possible for electrons that were produced by bacteria to be transported more quickly. Later,
voltage dropped as a result of the fact that all bacterial species ultimately reach the death
phase of their life cycle and that both electrodes began to operate less effectively after a
particular point in time. This result implies that the biocatalyst was only useful for a limit
voltage production of 30 days before reaching a state of low oxidation, at which time the
creation of electrons was at a low level. According to earlier studies, organic substrates
lead to a voltage drop during the operation. If the bacteria are to survive for a period
of time, the organic source should be enough to offer energy for their activities [21,40].
Because electrodes are what are utilized to transport the electrons, the two electrodes that
were employed in this investigation both exhibited great performance. During operation,
the capacity to generate electrons without experiencing a decreasing trend is contingent
on bacterial activity as well as the stability of organic substrates [41]. Figure 5 demon-
strates how the voltage trend increased gradually before starting to fall on the 30th day of
the MFC.
In order to shed light on the connection that exists between current and resistance in
MFCs, this study considered polarization behavior. To analyze the polarization study and
find the ideal current–resistance bond. On day 18, when the voltage was high, polarization
behavior was analyzed. On the other hand, a low exterior resistance suggested a high
voltage production but imprecise voltage stability, while a high external resistance indicated
a restricted capacity to generate voltage. At 1000 Ω in the GO anode, the maximum CD
was measured at 28.94 mA/m2 , whereas the highest PD was measured at 0.69 mW/m2 .
The internal resistance of the solution was measured at 581 Ω in that particular anode. The
GO showed a growing trend in CD as it went from higher to lower external resistance;
for instance, its high external resistance corresponded with 5.23 mA/m2 at 5000 Ω, and
28.94 mA/m2 at 100 Ω. In a manner analogous, the current density (CD) of the GO-ZnO
composite anode was high (38.15 mA/m2 ) when the external resistance was 1000 Ω and
the internal resistance effect was 497 Ω. Figure 6 demonstrates that the highest PD for
the composite anode was 1.05 mW/m2 . A comparative profile with previous literature is
presented in Table S1. Energy efficiency at the anode was lower because of the high external
resistance, but it may now be allowing electrons to flow to the cathode. In addition, since
electrons could not get through the bacteria to the anodes, the internal resistance was lower
than it would have been otherwise. Experiments on polarization have shown a pattern that
has been compared to this one before [42,43].
 Processes 2022, 10, x FOR PEER REVIEW 10 of 20

trend is contingent on bacterial activity as well as the stability of organic substrates [41].
Processes 2023, 11, 163 Figure 5 demonstrates how the voltage trend increased gradually before starting 10 of 19
to fall on
the 30th day of the MFC.

Processes 2022, 10, x FOR PEER REVIEW 11 of 20

Figure 5. Voltage generation trend through the MFC operation in the presence of self-fabricated
lower than it would havethe
been otherwise. Experiments on polarization have shown a pat-
Figure 5. Voltage generation trend through MFC operation in the presence of self-fabricated anodes.
anodes.
tern that has been compared to this one before [42,43].
In order to shed light on the connection that exists between current and resistance in
MFCs, this study considered polarization behavior. To analyze the polarization study and
find the ideal current–resistance bond. On day 18, when the voltage was high, polarization
behavior was analyzed. On the other hand, a low exterior resistance suggested a high
voltage production but imprecise voltage stability, while a high external resistance indi-
cated a restricted capacity to generate voltage. At 1000 Ω in the GO anode, the maximum
CD was measured at 28.94 mA/m2, whereas the highest PD was measured at 0.69 mW/m2.
The internal resistance of the solution was measured at 581 Ω in that particular anode. The
GO showed a growing trend in CD as it went from higher to lower external resistance; for
instance, its high external resistance corresponded with 5.23 mA/m2 at 5000 Ω, and 28.94
mA/m2 at 100 Ω. In a manner analogous, the current density (CD) of the GO-ZnO compo-
site anode was high (38.15 mA/m2) when the external resistance was 1000 Ω and the inter-
nal resistance effect was 497 Ω. Figure 6 demonstrates that the highest PD for the compo-
site anode was 1.05 mW/m2. A comparative profile with previous literature is presented
in Table
Figure
Figure 6. Polarization curves S1. Energy
6. Polarization
of GO efficiency
curves
and atanode
of GO and
GO-ZnO the anode
GO-ZnO wasthe
anode
during lower
during because of the high external re-
the operation.
operation.
sistance, but it may now be allowing electrons to flow to the cathode. In addition, since
electrons
Next, we could not get through
the rate the bacteriatransport
to the anodes, the internal
CV resistance
throughoutwas
Next, we determined thedetermined
rate of electron of electron
transport by performing
by performing CV throughout a
potential window range of 0.8 V in both positive and negative directions. The examination
a potential window of therange of 0.8 intervals
CV at various V in both positivetheand
demonstrated negative
bacterial biomass directions. The
catalytic activity ex-
that is
amination of the connected
CV at various intervals demonstrated the bacterial biomass
to the electrodes. The redox reactions that occurred throughout the process rep- catalytic
resented this
activity that is connected activity.
to the Figure 7 demonstrates
electrodes. the CV behavior
The redox reactions that atoccurred
different intervals
throughout while
the MFC is operating.
the process represented this activity. GO’s CV curves were scanned both forward
Figure 7 demonstrates the CV behavior at differ-(FS) and reversely (RS);
the FS was 1.0 × 10.0–5 mA on day 10, 2.6 × 10.0–5 mA on day 20 and 4.3 × 10.0–5 mA on day
ent intervals while the MFC is operating. GO’s CV curves were scanned both forward
30. However, the RS on days 10, 20, and 30 was –0.4 × 10.0–5 mA, 0.8 × 10.0–5 mA and –1.4
–5 –5
(FS) and reversely× 10.0 (RS); therespectively.
–5 mA, FS was 1.0 Days×10,10.0
20, andmA 30 wereonused
dayto10, 2.6 FS×and
test the 10.0
RS formA on
the GO-
day 20 and 4.3 ×ZnO –5
10.0composite;
mA on theday 30. However,
FS findings were 1.7 × 10.0the–6 mA,
RS on2.4 ×days
10.0–5 10,
mA and20, 5.9
and 30–5was
× 10.0 mA,
–0.4 × 10.0–5 mA, respectively,
0.8 × 10.0while –5 mA theand
RS values
–1.4 were
× 10.0 –5× mA,
–0.5 10.0–5 respectively.
mA, –1.8 × 10.0–5 Days
mA, and 10,–2.4
20,× 10.0
and–5
mA. The high RS for the GO-ZnO composite was –2.4 × 10.0–5 mA, while the FS was 5.9 ×
30 were used to test–5the FS and RS for the GO-ZnO composite; the FS findings were
10.0 mA.–5 The FS or RS may be used to depict the oxidation and reduction rates at various
1.7 × 10.0–6 mA, 2.4times
× 10.0 mA and
throughout the MFC’s 10.0–5 mA,
5.9 ×operation. Dayrespectively,
30 found strong while the RS
oxidation values
peaks were
for both GO
–0.5 × 10.0–5 mA, and –1.8the 10.0–5 mA,
×GO-ZnO and –2.4
composite, × 10.0
and that
–5 mA. The high RS for the GO-ZnO
day reverse electron transport accounted for most
composite was –2.4 × 10.0
of the –5 mA,
decrease in rate. On day
while the30,
FSoxygen
was 5.9 in the 10.0–5 mA.
× cathode chamber Theincreased
FS or RS electron
maydis-be
charge and neutralized electrons before reaching the
used to depict the oxidation and reduction rates at various times throughout the MFC’s anode, boosting oxidation and re-
duction. GO’s biological process took longer than GO because in ZnO, there were no oxi-
operation. Day 30dation
foundorstrong
reduction oxidation
currents on peaks
the 10thforday,
both GO to
leading andpoor the GO-ZnO
metal composite,
ion degradation. The
and that day reverse electron transport accounted for most of the
analysis of the pattern of biofilm formation at the GO and GO-ZnO composite decrease in rate. On
anodes
during the reaction took into account a wide range of other factors. Utilizing the CV curve
and the corresponding Equation 1, Cp may be determined. Biofilm strength during the
reaction was reflected by the high Cp values. If the Cp value was high, the biofilm ap-
peared to be well formed and stable. On day 30, GO displays 0.00006 F/g, whereas GO-
ZnO displays 0.00009 F/g. The results of these and other studies provide an explanation
for and suggest the presence of substantial biofilm growth. It has been observed that sim-
Processes 2023, 11, 163 11 of 19

day 30, oxygen in the cathode chamber increased electron discharge and neutralized elec-
trons before reaching the anode, boosting oxidation and reduction. GO’s biological process
Processes 2022, 10, x FOR PEER REVIEW
took longer than GO because in ZnO, there were no oxidation or reduction currents 12 ofon
20

the 10th day, leading to poor metal ion degradation. The analysis of the pattern of biofilm
formation at the GO and GO-ZnO composite anodes during the reaction took into account
aTable
wide2.range of other
Cp values of GOfactors. Utilizing
and GO-ZnO theCV
anode CVcurves.
curve and the corresponding Equation (1),
Cp may be determined. Biofilm strength during the reaction was reflected by the high Cp
values.Days
If the Cp value was high, the biofilm Specific Capacitance
appeared (F/g)formed and stable. On
to be well
GO
day 30, GO displays 0.00006 F/g, whereas GO-ZnO displays 0.00009 GO-ZnO
F/g. The results of
these and10other studies provide 0.00001 0.00004 of substantial
an explanation for and suggest the presence
20
biofilm growth. 0.00004that similar patterns exist for
It has been observed 0.00005
other materials [44].
Other days
30 results are shown in Table
0.000062. 0.00009

Figure 7.
Figure 7. CV
CV study
study of
of the
the GO
GO and
and GO-ZnO.
GO-ZnO.

3.3. Durability
Table Study
2. Cp values of GO
of GO and and GO-ZnO
GO-ZnO Electrode
anode CV curves.
The durability of MFC electrodes is important for energy generation and wastewater
treatment. The present study examined the anodes’ Specific Capacitance
stability (F/g)
(biocompatibility and dura-
Days
bility). SEM images of anode biofilm after 30 GOdays of MFC operation showed GO-ZnObiocompat-
ible and stable10anodes. Durability was tested by cycling the anodes three
0.00001 times at 1000 Ω.
0.00004
Figure 5 reveals 20
that the high voltage for GO
0.00004
was observed on the 18th day, while the
0.00005
maximum voltage for GO-ZnO was achieved on the same day. The sudden increasing
30 0.00006 0.00009
trend in voltage could be attributed to the introduction of new inoculum as well as the
quick electron transfer. After that phase, the voltage continued to drop in two scenarios,
3.3.
and Durability
the GO-ZnO Study of GO
anode and GO-ZnO
revealed a moreElectrode
gradual decline than the GO anode. The GO an-
ode wasThe the controlofinMFC
durability bothelectrodes
scenarios. isThis indicates
important forthat the GO-ZnO
energy generation anode provided a
and wastewater
surface area
treatment. Thethat was more
present studyconducive
examinedto thethe growth
anodes’ of bacteria
stability in comparison
(biocompatibility andto durabil-
the GO
ity).
anode.SEM Theimages
biofilm ofthat
anodewasbiofilm
found after 30 days had
on GO-ZnO of MFC operation
a higher densityshowed
than the biocompatible
biofilm that
and
was stable
detectedanodes.
on GO.Durability was testedthat
This demonstrated by the
cycling the anodes
continual feeding three 1000 Ω.
times atsubstrate
of organic
Figure 5 reveals that the high voltage for GO was observed on the
gave the bacterial colony the ability to produce electrons. Both electrodes demonstrated18th day, while the
maximum voltage for GO-ZnO
constant performance was achieved
throughout on thecycles,
the operating same day. withThe
nosudden
evidence increasing trend
of biofilm or
in voltage
other could
material be attributed
being destroyed.toThe theperformance
introductionof ofthe
new inoculum
GO-ZnO as wellwas
electrode as the quick
superior
electron
to that of transfer.
the GOAfter that phase,
electrode due to thethe
voltage continued
fact that to drop in two
ZnO contributed scenarios, electron
to enhanced and the
GO-ZnO
transit and anode revealed
minimized a more
the buildupgradual decline that
of elements thanwere
the GO notanode. The GO
acceptable. anode was
Nevertheless,
the
bothcontrol
producedin both scenarios.
anodes exhibitedThis indicates
excellent that the for
endurance GO-ZnO
a periodanode
of 30provided
days. Several a surface
stud-
area that was more conducive to the growth of bacteria in comparison
ies also investigated the durability via electrochemical methods of the prepared electrodes to the GO anode.
The
duringbiofilm that was
continuous found on
operation GO-ZnO
[45–47]. hadother
On the a higher
hand, density than theetbiofilm
Senthilkumar al. [48] that was
revealed
detected on GO. This demonstrated that the continual feeding of organic
that waste-derived anodes had a durability span of 15 days. Table 3 provides a compari- substrate gave
the
sonbacterial colonysystems
of the several the ability
thattohave
produce
beenelectrons.
suggested. Both electrodes demonstrated constant
Processes 2023, 11, 163 12 of 19

performance throughout the operating cycles, with no evidence of biofilm or other material
being destroyed. The performance of the GO-ZnO electrode was superior to that of the GO
electrode due to the fact that ZnO contributed to enhanced electron transit and minimized
the buildup of elements that were not acceptable. Nevertheless, both produced anodes
exhibited excellent endurance for a period of 30 days. Several studies also investigated
the durability via electrochemical methods of the prepared electrodes during continuous
operation [45–47]. On the other hand, Senthilkumar et al. [48] revealed that waste-derived
anodes had a durability span of 15 days. Table 3 provides a comparison of the several
systems that have been suggested.

Table 3. A profile comparison of prepared anodes with previous work.

Anode Electrode Inoculation Durability (Days) Current Density (mA/m2 ) References

Mix culture
Fe3 O4 /Cp e 16.5 - [49]
bacteria species
Cp e /HAPNWf Shewanella loihica 2.1 2.90 mA/cm2 [50]
Fe3 O4 /AC 1/ SSM 2 Pre-acclimated effluent 5.0 1.90 mA/m2 [51]
AC 1 /PANI 3 /D-CSA 4 / GF 5 Mix culture bacteria 6.2 110 mA/m2 [52]
CNT 6 /CC 7 Mix culture bacteria 8–9 65 mA/m2 [53]
NGNS 8 Escherichia coli 4.1 - [54]
Graphene/PEDOT 9/ Cp 10 Escherichia coli 24 3.59 mA/m2 [55]
PEI 11 /GF 12 Saccharomyces cerevisiae 5 - [56]
GO Mix culture bacteria 30 28.94 mA/m2 Present study
GO-ZnO Mix culture bacteria 30 38.15789 mA/m2 Present study
1Activated carbon. Stainless steel meshes. Polyaniline. D-camforsulfonic acid. Graphite felt. 6 Carbon
2 3 4 5

nanotubes. 7 Carbon cloth. 8 N-doped graphene nanosheets. 9 Poly(3,4-ethylenedioxythiophene). 10 Carbon

paper. 11 Polyethyleneamine. 12 Hydroxylated and aminated polyaniline nanowire.

3.4. Degradation and Biological Studies

The most recent method for expanding MFC applications is to incorporate MFCs into
the degradation of inorganic contaminants in water. For this reason, we set out to utilize
MFCs to get rid of the bulk of the Cd2+ , Cr3+ , Pb2+ , and Ni2+ in synthetic wastewater, since
their presence might induce diseases that can be lethal to people. The bacterial species sur-
rounding the electrodes were fed by the food waste, and the biofilm was solely accountable
for the high rate of degradation. According to Table 4, the electrodes significantly aid in
degradation due to the thick biofilm that coats the anodes. The GO-ZnO composite anode
was superior to GO in terms of degradation efficiency (81.20% vs. 92.71%). The degradation
efficiency of the GO-ZnO composite increased in both situations. ZnO demonstrated better
biocompatibility than GO did. However, the organic substrate ran out of energy too quickly
for the bacteria to continue respiration after being used for a while. Bacterial species entered
the death phase due to a lack of organic substrate, which also inhibited the metal ions from
transforming into an insoluble form. Previous research by Singh et al. [57] examined the
impacts of different organic sources and concluded that a deficiency of access to such a
source might reduce the effectiveness of MFCs. That said, although the organic sources
used are sufficient for bacterial activity in the sense of metal ion degradation, more force
is necessary for high electricity production. For the sole purpose of monitoring metal
concentrations, AAS has no further applications. Previous studies have shown that once an
MFC operation is performed, the soluble metal ions become insoluble (sludge develops).
The mechanisms of MFCs (electrons generated and transported) have been well described
in prior research [26,58]. In an electrochemical reaction, soluble metal ion forms are reduced
to solids by accepting electrons created throughout the process (precipitates) [26,58].
Processes 2023, 11, 163 13 of 19

Table 4. Overview of the degradation of metal ions with the GO and GO-ZnO via MFCs.
Processes 2022, 10, x FOR PEER REVIEW 14 of 20

Electrode
Organic Matter Concentration of Metal Ions Operational Days Degradation Efficiency (%)
Anode Cathode
Table 4. Overview of the degradation of metal ions with the GO and GO-ZnO via MFCs.
0 0.00
Organic Concentration Electrode Operational
10
Degradation
40.45
Matter of MetalGO
Ions Graphite rod
Anode Cathode Days Efficiency (%)
20 76.10
0 0.00
Food waste 10 30 81.20
10 40.45
GO Graphite rod 0 20 0.00
76.10
10 30 43.89
81.20
Food waste 10
GO-ZnO Graphite rod
20 0 0.00
79.04
10 43.89
GO-ZnO Graphite rod 30 92.71
20 79.04
30 92.71
The anode-treated surfaces were analyzed by SEM to determine the kind of bacterial
coloniesThethatanode-treated
had colonized themwere
surfaces (Figure 8). After
analyzed 30 days,
by SEM many distinct
to determine the kindbacterial colonies
of bacterial
were
colonies that had colonized them (Figure 8). After 30 days, many distinct bacterial colonies they
seen on the surface of the treated anodes (GO and GO-ZnO composite), and
were
werethought
seen ontothebesurface
responsible
of the for the anodes
treated generation of energy
(GO and GO-ZnO and metal ionand
composite), degradation.
they
were thought to be responsible for the generation of energy and metal
The findings suggested that a controlled amount of biofilm growth is beneficial to power ion degradation.
The findings
production. suggested
Sasaki that aalso
et al. [59] controlled
showed amount
that a of biofilm
small growth
amount ofisbiofilm
beneficial
onto thepower
surface of
production. Sasaki et al. [59] also showed that a small amount of
anodes improves energy generation and degradation rates. The composition of the EPS biofilm on the surface of was
anodes improves energy generation and degradation rates. The composition
impacted by the increased biomass on the surface of the anode, which resulted in decreased of the EPS
was impacted by the increased biomass on the surface of the anode, which resulted in
energy generation. Therefore, it seems that MFCs result in clean energy technologies that do
decreased energy generation. Therefore, it seems that MFCs result in clean energy tech-
not release harmful emissions or waste products. Furthermore, in accordance with previous
nologies that do not release harmful emissions or waste products. Furthermore, in accord-
studies
ance [60,61], the treated
with previous studiesanodes’
[60,61], SEM imageanodes’
the treated showed rod-shaped
SEM image showedfilament appendages,
rod-shaped
which indicates
filament the existence
appendages, of conductive
which indicates pili types
the existence of bacterial
of conductive species.
pili types This means
of bacterial
thatspecies.
they used conductive
This means piliused
that they to transfer the pili
conductive electrons to the
to transfer theanode electrode.
electrons The most
to the anode
prominent
electrode.conductive pili types
The most prominent of bacterial
conductive species
pili types are Escherichia,
of bacterial species areG. bacter, Klebsiella,
Escherichia, G.
Bacillus,
bacter,and Actinobacillus.
Klebsiella, Therefore,
Bacillus, and the present
Actinobacillus. SEMthe
Therefore, images
presentandSEMprevious
imagesevidence
and pre- show
thatvious evidence
there show
is a great that therethat
possibility is a great
therepossibility that there
are conductive aretypes
pili conductive pili types
of bacterial of
species, as
bacterial species,
mentioned above. as mentioned above.

Figure 8. SEM
Figure images
8. SEM imagesofoftreated
treated anodes.
anodes.

3.5. Oxidation of Food Waste

The action of microorganisms is necessary for the high performance of MFCs in the
transportation of electrons and the degradation of contaminants. In the scientific litera-
ture [60,61], several microbial species, such as Escherichia, G. bacter, Klebsiella,
 3.5. Oxidation of Food Waste
The action of microorganisms is necessary for the high performance of MFCs in the
Processes 2023, 11, 163 transportation of electrons and the degradation of contaminants. In the scientific literature 14 of 19
[60,61], several microbial species, such as Escherichia, G. bacter, Klebsiella, Bacillus, and Ac-
tinobacillus, have been revealed to be exoelectrogens. In this experiment, several species of
microorganisms were grown have
Bacillus, and Actinobacillus, on food waste,
been which
revealed served
to be as an organic
exoelectrogens. Insubstrate. Food
this experiment,
waste was a disaccharide in its original form. However, it was subsequently changed
several species of microorganisms were grown on food waste, which served as an organic into
glucose and then oxidized by bacteria, which caused the waste to produce electrons.
substrate. Food waste was a disaccharide in its original form. However, it was subse- The
investigation’s
quently changed findings on the –processes
into glucose are summarized
and then oxidized as follows
by bacteria, which(Equations
caused the(3)–(5)):
waste to
produceOxidation:
electrons. The investigation’s
Food waste findings on the –processes
C6H12O6 + 6H2O are summarized
6CO2 + follows
as
(Equations (3)–(5)): (3)
24H+ + 24e−
+ −
Oxidation: Food24H
Reduction: waste → −C+6 H
+ + 24e 6O122O6 + 6H2 O → 6CO2 + 24H12H
+ 24e
2O (4)(3)
General: C6H 12O6 + 6O2 24H+ + 24e− + 6O 6CO 2 + 6H
Reduction: 2 → 12H 2 O 2O + Electricity (5)(4)
Moving electrons from C
General: the
6 Hanode
12 O6 +electrode
6O2 → 6CO(where they
2 + 6H 2 Oare generated) to the cathode(5)
+ Electricity
electrode (where they are reduced) is a key step in the oxidation process. Since there was
Moving electrons from the anode electrode (where they are generated) to the cathode
only a single chamber in the MFC employed in this work, moving the protons from the
electrode (where they are reduced) is a key step in the oxidation process. Since there was
anode to the cathode chamber was significantly easier [62,63]. The electrons are trans-
only a single chamber in the MFC employed in this work, moving the protons from the
ferred via the provided outer circuit that is located outside. Before electrons can be moved
anode to the cathode chamber was significantly easier [62,63]. The electrons are transferred
to the cathode electrode in bacterial cells, they must first go to the cathode electrode
via the provided outer circuit that is located outside. Before electrons can be moved to the
through the anode electrode. A systematic presentation has been shown in Figure 9, it was
cathode electrode in bacterial cells, they must first go to the cathode electrode through the
modified according to the present study to show how electrons are generated and trans-
anode electrode. A systematic presentation has been shown in Figure 9, it was modified
ferred to the anode electrode.
according to the present study to show how electrons are generated and transferred to the
anode electrode.

Figure 9. Food waste microbiologically oxidized in a single-chamber MFC (figure modified from [64]
Figure 9. Food waste microbiologically oxidized in a single-chamber MFC (figure modified from
with Elsevier’s permission).
[64] with Elsevier’s permission).
3.6. Assessment of Food Waste as an Organic Substrate
When trying to maximize the performance of MFCs, among the most important factors
to take into account is the organic substrate. Numerous studies have produced distinctive
findings by making use of a wide variety of natural organic substrates [65]. Any source
of carbohydrate that is used as an organic substrate in the process of operating MFCs has
the potential to provide bacteria with the energy necessary to carry out their oxidative
respiration activity. However, in the previous research that has been conducted, there have
only been a very small number of attempts to use natural organic substrates in MFCs [66].
Processes 2023, 11, 163 15 of 19

These substrates include rotten rice, biomass waste, vegetables, chocolate waste, and fruits.
Among these researchers, Salvin et al. [67] used mangroves as an organic substrate in MFCs
in order to increase the devices’ energy performance. In contrast, food waste was used as an
organic substrate in our experiment alongside glucose. This was performed to examine the
relative performance of the food waste in comparison to the other commercial substrates
even though its carbohydrate-based organic substrates contain a maximum concentration
of bacterial nutrients than food waste. Table 5 summarizes the performance of all of the
organic substrates, and it reveals that the best performance was achieved by the food waste.
This means that glucose is a simple sugar that is easily and rapidly oxidized, which reduces
the operation’s durability as well as the transportation rate.

Table 5. Performance of commercial glucose and food waste as an organic substrate.

Electrode Organic Substrate Voltage (mV) Operation Duration (Days) Degradation Efficiency (%)
GO Glucose 24 10 18.00
GO-ZnO Glucose 38 10 31.20
GO Food waste 49 10 40.45
GO-ZnO Food waste 64 10 43.89

4. Comparative Profile with Pervious Literature

A recent subject in MFCs is the preparation of the high-quality, low-cost base material
for anode fabrication. The development of anodes considers several variables to improve
their conductivity, durability, mechanical, thermal, and chemical biocompatibility toward
the operating microorganism in MFCs. The current findings indicate that both anodes
exhibit high mechanical, chemical, and biological compatibility as described in the pre-
ceding sections. The chemical compatibility for bacterial growth is evident in the SEM
photos of the biofilm around both anodes. Similarly, Figure 5 shows the rate of electron
transit, demonstrating the good results. Both anodes were found to last 30 days when used
continuously. Within 18 days, each electrode produced a maximum voltage of 110 mV (GO)
and 130 mV (GO-ZnO). According to the most recent research, as shown in Table 3, the rate
of electron transmission from anode to cathode was determined to be excellent. The highest
CD was 28.94 mA/m2 (GO) and 38.15 mA/m2 (GO-ZnO), while the PD was 0.692 mW/m2
(GO) and 1.059 mW/m2 (GO-ZnO), respectively. The maximum CD and PD indicated that
electron transportation was efficient for 30 days, which is consistent with the longevity
of the anodes. Several studies used the same strategy to explain the durability and com-
parative profile of the self-fabricated anodes [7,8,45,46,54]. For example, Senthil et al. [48]
fabricated the waste-derived 3D-carbon aerogel-ceria-nitrogen-doped reduced graphene
oxide-based anode electrode. Their anode was advertised as having a 360-h continuous
batch life. Similar to this, Table S2 presents a comparative profile on the degradation of
toxic metal. In comparison to earlier literature, the current archival degradation efficiency
was 92.71%, which is much higher. The presence of a conductive, biocompatible electrode
and a very advantageous organic substrate in the form of food waste is what accounts for
the high degradation efficiency and energy generation/transport.

Present Challenges and Future Perfectives

The potential for MFCs to be employed in a wide range of applications, including the
bioremediation of wastewater, is now receiving a lot of attention [68]. Despite this, MFCs
are still struggling to overcome limitations brought on by the design and composition of the
electrodes. The generation of highly conductive electrodes and subsequent modifications
to those electrodes have the potential to raise the standing of MFC technology in the
context of the biodegradation of contaminants and the treatment of wastewater. The MFC
is a revolutionary technology that could provide clean, safe, sustainable energy and help
preserve the environment [40]. However, MFCs are a very new topic of research in the
Processes 2023, 11, 163 16 of 19

scientific world; for this reason, it will need a considerable amount of effort and time
before it can be regarded as commercially feasible. The developing problem with MFCs is
that the organic substrate and electrode material are both prone to instability [21]. In the
current investigation, favorable outcomes were achieved; nevertheless, it was challenging
to maintain the system throughout the course of the whole research project. The scaling
up of MFCs requires a long-lasting organic substrate. Recently, food waste has been
used as a substrate in MFCs, which has extended the duration of the process to 30 days.
To contemplate the scaling up of MFC dimensions, the requirements for a functioning
organic substrate over the long term include a high level of stability as well as a significant
quantity of carbohydrates. Another crucial aspect of the MFC that must be taken into
account is the material that will be used for the electrodes. The MFC was not capable of
producing an appropriate quantity of energy because of the material that was used for the
electrodes. Anode material should transfer electrons effectively and create a biocompatible
environment for bacteria to grow a biofilm on the anode surface [69,70]. Due to their
affordability and high performance, electrode materials made from biomass have attracted
a lot of interest. Many studies have already been conducted on the conversion of biomass
to produce the electrodes [71]. A potentially fruitful strategy for the future may include the
alteration of material obtained from biomass using polymers coupled with metal oxide.

5. Conclusions
This article shows how waste derived MFC anodes transport the electrons and degrade
the hazardous metal ions in synthetic wastewater. Our results show that an anode made
from oil palm tree material may improve electron transport with GO and GO-ZnO, leading
to superior energy performance. The MFC’s use of waste food as an organic source is an
ideal situation. A little study on the use of natural organic substrates has investigated the
effectiveness of MFCs in producing electricity and treating wastewater. In our research,
it was found that the prepared electrodes were extraordinarily biocompatible and stable
for the operation of MFCs, as shown by several material characterizations and results. GO
and GO-ZnO anodes generated a solid biofilm, proving their safety and chemical stability
for 30 days. The metal ion degradation also occurred, and significant progress was made
in a short amount of time. As an organic substrate, food wastes were used to energize
the bacterial activities in MFCs. It took 30 days to finish each MFC operation, but only
18 days to reach peak energy efficiency. The composite anode was superior to the GO in
terms of energy efficiency and electron transport. The MFC’s organic substrate was still not
highly stable, but the produced electrodes improved electron transportation. Therefore,
future studies should concentrate on the durability of organic substrates in MFCs at the
industrial scale.

Supplementary Materials: The following supporting information can be downloaded at: https:
//www.mdpi.com/article/10.3390/pr11010163/s1, Table S1: Comparative analysis of previously
published research and the current work in MFC. Table S2: A comparison of the degradation of metal
ions using various anodes in MFC.
Author Contributions: A.A., conceptualization, methodology, writing—original draft preparation,
visualization, investigation, material synthesis and characterizations. M.B.A. and M.N.M.I. results
interpretation and writing—reviewing and editing. A.A., supervision and funding acquisition. This
article has been read and approved by all listed authors. All authors have read and agreed to the
published version of the manuscript.
Funding: The authors extend their appreciation to the Deputyship for Research & Innovation,
Ministry of Education in Saudi Arabia for funding this research work through the project number
(IF2/PSAU/2022/01/20629).
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: All the data have been included in the text.
Processes 2023, 11, 163 17 of 19

Acknowledgments: The authors extend their appreciation to the Deputyship for Research & Inno-
vation, Ministry of Education in Saudi Arabia for funding this research work through the project
number (IF2/PSAU/2022/01/20629).
Conflicts of Interest: The authors have stated that there are no competing interests that might
influence their work.

References
1. Heilmann, J.; Logan, B.E. Production of electricity from proteins using a microbial fuel cell. Water Environ. Res. 2006, 78, 531–537.
[CrossRef] [PubMed]
2. Logan, B.E.; Regan, J.M. Microbial fuel cells—Challenges and applications. Environ. Sci. Technol. 2006, 40, 5172–5180. [CrossRef]
3. Khatoon, A.; Mohd Setapar, S.H.; Parveen, T.; Rafatullah, M. Outlook on the role of microbial fuel cells in remediation of
environmental pollutants with electricity generation. Catalysts 2020, 10, 819.
4. Parkhey, P.; Sahu, R. Microfluidic microbial fuel cells: Recent advancements and future prospects. Int. J. Hydrogen Energy 2021, 46,
3105–3123. [CrossRef]
5. Choudhury, P.; Ray, R.N.; Bandyopadhyay, T.K.; Basak, B.; Muthuraj, M.; Bhunia, B. Process engineering for stable power recovery
from dairy wastewater using microbial fuel cell. Int. J. Hydrogen Energy 2021, 46, 3171–3182. [CrossRef]
6. Ahmad, H.; Parveen, T.; Ahmad, A.; Oves, M.; Ismail, I.M.; Qari, H.A. Recent advances in metal decorated nanomaterials and
their various biological applications: A review. Front. Chem. 2020, 8, 341.
7. Kirubaharan, C.J.; Yoo, D.J.; Kim, A.R. Graphene/poly (3, 4-ethylenedioxythiophene)/Fe3 O4 nanocomposite–An efficient oxygen
reduction catalyst for the continuous electricity production from wastewater treatment microbial fuel cells. Int. J. Hydrogen Energy
2016, 41, 13208–13219.
8. Kirubaharan, C.J.; Kumar, G.G.; Sha, C.; Zhou, D.; Yang, H.; Nahm, K.S.; Raj, B.S.; Zhang, Y.; Yong, Y.-C. Facile fabrication of
Au@ polyaniline core-shell nanocomposite as efficient anodic catalyst for microbial fuel cells. Electrochim. Acta 2019, 328, 135136.
[CrossRef]
9. Han, D.; Wu, S.; Zhang, S.; Deng, Y.; Cui, C.; Zhang, L.; Long, Y.; Li, H.; Tao, Y.; Weng, Z. A Corrosion-Resistant and Dendrite-Free
Zinc Metal Anode in Aqueous Systems. Small 2020, 16, 2001736. [CrossRef]
10. Rodríguez-Couto, S. Development and modification of materials to build cost-effective anodes for microbial fuel cells (MFCs):
An overview. Biochem. Eng. J. 2020, 164, 107779.
11. Idris, M.O.; Noh, N.A.M.; Yaqoob, A.A. Sustainable microbial fuel cell functionalized with a bio-waste: A feasible route to
formaldehyde bioremediation along with bioelectricity generation. Chem. Eng. J. 2022, 1, 140781.
12. Idris, M.O.; Guerrero-Barajas, C.; Kim, H.-C. Scalability of biomass-derived graphene derivative materials as viable anode
electrode for a commercialized microbial fuel cell: A systematic review. Chin. J. Chem. Eng. 2022, in press.
13. Fu, C.; Zhao, G.; Zhang, H.; Li, S. Evaluation and characterization of reduced graphene oxide nanosheets as anode materials for
lithium-ion batteries. Int. J. Electrochem. Sci. 2013, 8, 6269–6280.
14. Kurra, N.; Jiang, Q.; Nayak, P.; Alshareef, H.N. Laser-derived graphene: A three-dimensional printed graphene electrode and its
emerging applications. Nano Today 2019, 24, 81–102. [CrossRef]
15. Thepsuparungsikul, N.; Ng, T.; Lefebvre, O.; Ng, H. Different types of carbon nanotube-based anodes to improve microbial fuel
cell performance. Water Sci. Technol. 2014, 69, 1900–1910. [CrossRef] [PubMed]
16. Cai, T.; Meng, L.; Chen, G.; Xi, Y.; Jiang, N.; Song, J.; Zheng, S.; Liu, Y.; Zhen, G.; Huang, M. Application of advanced anodes in
microbial fuel cells for power generation: A review. Chemosphere 2020, 248, 125985. [CrossRef]
17. Chiu, W.-T.; Chang, T.-F.M.; Sone, M.; Tixier-Mita, A.; Toshiyoshi, H. Roles of TiO2 in the highly robust Au nanoparticles-TiO2
modified polyaniline electrode towards non-enzymatic sensing of glucose. Talanta 2020, 212, 120780. [CrossRef]
18. Deng, C.; Ma, C.; Lau, M.L.; Skinner, P.; Liu, Y.; Xu, W.; Zhou, H.; Ren, Y.; Yin, Y.; Williford, B. Amorphous and crystalline TiO2
nanoparticle negative electrodes for sodium-ion batteries. Electrochim. Acta 2019, 321, 134723. [CrossRef]
19. Baratto, C.; Ferroni, M.; Rocco, D.; De Angelis, C.; Faglia, G. Chemical sensing with ZnO nano and microwires. Nano-M&D 2019,
2019, 28.
20. Shakeel, N.; Ahamed, M.I.; Kanchi, S.; Kashmery, H.A. Green synthesis of ZnO nanoparticles decorated on polyindole
functionalized-Mcnts and used as anode material for enzymatic biofuel cell applications. Sci. Rep. 2020, 10, 5052.
21. Fadzli, F.S.; Bhawani, S.A.; Adam Mohammad, R.E. Microbial fuel cell: Recent developments in organic substrate use and
bacterial electrode interaction. J. Chem. 2021, 2021, 4570388. [CrossRef]
22. Rafatullah, M.; Chua, Y.S.; Umar, K. Recent advances in anodes for microbial fuel cells: An overview. Materials 2020, 13, 2078.
23. Ghisellini, P.; Ulgiati, S. Circular economy transition in Italy. Achievements, perspectives and constraints. J. Clean. Prod. 2020,
243, 118360. [CrossRef]
24. Paulchamy, B.; Arthi, G.; Lignesh, B. A simple approach to stepwise synthesis of graphene oxide nanomaterial. J. Nanomed.
Nanotechnol. 2015, 6, 1.
25. Fahmey, M.S.; El-Aassar, A.-H.M.; Abo-Elfadel, M.M.; Orabi, A.S.; Das, R. Comparative performance evaluations of nanomaterials
mixed polysulfone: A scale-up approach through vacuum enhanced direct contact membrane distillation for water desalination.
Desalination 2019, 451, 111–116. [CrossRef]
Processes 2023, 11, 163 18 of 19

26. Yaakop, A.S.; Ahmad, A. Application of microbial fuel cells energized by oil palm trunk sap (OPTS) to remove the toxic metal
from synthetic wastewater with generation of electricity. Appl. Nanosci. 2021, 11, 1949–1961.
27. Timmers, R.A.; Strik, D.P.; Hamelers, H.V.; Buisman, C.J. Characterization of the internal resistance of a plant microbial fuel cell.
Electrochim. Acta 2012, 72, 165–171. [CrossRef]
28. Igboamalu, T.E.; Bezuidenhout, N.; Matsena, M.T.; Chirwa, E. Microbial fuel cell power output and growth: Effect of pH on
anaerobic microbe consortium. Chem. Eng. Tran. 2019, 76, 1–16.
29. Katuri, K.P.; Scott, K.; Head, I.M.; Picioreanu, C.; Curtis, T.P. Microbial fuel cells meet with external resistance. Bioresour. Technol.
2011, 102, 2758–2766. [CrossRef]
30. Yaqoob, A.A.; Yaakop, A.S. Modified graphene oxide anode: A bioinspired waste material for bioremediation of Pb2+ with energy
generation through microbial fuel cells. Chem. Eng. J. 2021, 417, 128052. [CrossRef]
31. Durmus, Z.; Kurt, B.Z.; Durmus, A. Synthesis and characterization of graphene oxide/zinc oxide (GO/ZnO) nanocomposite and
its utilization for photocatalytic degradation of basic Fuchsin dye. ChemistrySelect 2019, 4, 271–278. [CrossRef]
32. Anandhavelu, S.; Thambidurai, S. Single step synthesis of chitin/chitosan-based graphene oxide–ZnO hybrid composites for
better electrical conductivity and optical properties. Electrochim. Acta 2013, 90, 194–202. [CrossRef]
33. Chen, C.-H.; Chung, T.-Y.; Shen, C.-C.; Yu, M.-S.; Tsao, C.-S.; Shi, G.-N.; Huang, C.-C.; Ger, M.-D.; Lee, W.-L. Hydrogen storage
performance in palladium-doped graphene/carbon composites. Int. J. Hydrogen Energy 2013, 38, 3681–3688. [CrossRef]
34. Feng, J.-X.; Zhang, Q.-L.; Wang, A.-J.; Wei, J.; Chen, J.-R.; Feng, J.-J. Caffeine-assisted facile synthesis of platinum@ palladium
core-shell nanoparticles supported on reduced graphene oxide with enhanced electrocatalytic activity for methanol oxidation.
Electrochim. Acta 2014, 142, 343–350. [CrossRef]
35. Bikkarolla, S.K.; Cumpson, P.; Joseph, P.; Papakonstantinou, P. Oxygen reduction reaction by electrochemically reduced graphene
oxide. Faraday Discuss. 2014, 173, 415–428. [CrossRef] [PubMed]
36. Li, X.; Sun, P.; Yang, T.; Zhao, J.; Wang, Z.; Wang, W.; Liu, Y.; Lu, G.; Du, Y. Template-free microwave-assisted synthesis of ZnO
hollow microspheres and their application in gas sensing. CrystEngComm 2013, 15, 2949–2955. [CrossRef]
37. Umar, K.; Adnan, R.; Rashid, M. Graphene oxide–ZnO nanocomposite: An efficient visible light photocatalyst for degradation of
rhodamine B. Appl. Nanosci. 2021, 11, 1291–1302.
38. Serrà, A. Self-assembled oil palm biomass-derived modified graphene oxide anode: An efficient medium for energy transportation
and bioremediating Cd (II) via microbial fuel cells. Arab. J. Chem. 2021, 14, 103121.
39. Rodríguez-Couto, S.; Ahmad, A. Preparation, characterization, and application of modified carbonized lignin as an anode for
sustainable microbial fuel cell. Process Saf. Environ. Prot. 2021, 155, 49–60.
40. Yaqoob, S.B.; Bhawani, S.A.; Ismail Abdulrahman, R.M. Utilization of Mangifera indica as Substrate to Bioremediate the Toxic
Metals and Generate the Bioenergy through a Single-Chamber Microbial Fuel Cell. J. Chem. 2021, 2021, 8552701. [CrossRef]
41. Daud, N.N.M. Application of rotten rice as a substrate for bacterial species to generate energy and the removal of toxic metals
from wastewater through microbial fuel cells. Environ. Sci. Pollut. Res. 2021, 28, 62816–62827. [CrossRef] [PubMed]
42. Sajana, T.; Ghangrekar, M.; Mitra, A. Application of sediment microbial fuel cell for in situ reclamation of aquaculture pond water
quality. Aquac. Eng. 2013, 57, 101–107. [CrossRef]
43. Sajana, T.; Ghangrekar, M.; Mitra, A. Effect of presence of cellulose in the freshwater sediment on the performance of sediment
microbial fuel cell. Bioresour. Technol. 2014, 155, 84–90. [CrossRef] [PubMed]
44. Fadzli, F.S.; Rashid, M. Electricity generation and heavy metal remediation by utilizing yam (Dioscorea alata) waste in benthic
microbial fuel cells (BMFCs). Biochem. Eng. J. 2021, 172, 108067. [CrossRef]
45. Senthilkumar, N.; Pannipara, M.; Al-Sehemi, A.G. PEDOT/NiFe2 O4 nanocomposites on biochar as a free-standing anode for
high-performance and durable microbial fuel cells. New J. Chem. 2019, 43, 7743–7750. [CrossRef]
46. Gnana Kumar, G.; Kirubaharan, C.J.; Udhayakumar, S.; Karthikeyan, C.; Nahm, K.S. Conductive polymer/graphene supported
platinum nanoparticles as anode catalysts for the extended power generation of microbial fuel cells. Ind. Eng. Chem. Res. 2014, 53,
16883–16893. [CrossRef]
47. Yaakop, A.S.; Rafatullah, M. Utilization of biomass-derived electrodes: A journey toward the high performance of microbial fuel
cells. Appl. Water Sci. 2022, 12, 99–112.
48. Senthilkumar, N.; Aziz, M.A.; Pannipara, M.; Alphonsa, A.T.; Al-Sehemi, A.G.; Balasubramani, A. Waste paper derived three-
dimensional carbon aerogel integrated with ceria/nitrogen-doped reduced graphene oxide as freestanding anode for high
performance and durable microbial fuel cells. Bioprocess Biosyst. Eng. 2020, 43, 97–109. [CrossRef]
49. Kim, J.R.; Min, B.; Logan, B.E. Evaluation of procedures to acclimate a microbial fuel cell for electricity production. Appl. Microbiol.
Biotechnol. 2005, 68, 23–30. [CrossRef]
50. Zhao, Y.; Nakanishi, S.; Watanabe, K.; Hashimoto, K. Hydroxylated and aminated polyaniline nanowire networks for improving
anode performance in microbial fuel cells. J. Biosci. Bioeng. 2011, 112, 63–66. [CrossRef]
51. Peng, X.; Yu, H.; Ai, L.; Li, N.; Wang, X. Time behavior and capacitance analysis of nano-Fe3O4 added microbial fuel cells.
Bioresour. Technol. 2013, 144, 689–692. [CrossRef] [PubMed]
52. Scott, K.; Rimbu, G.; Katuri, K.; Prasad, K.; Head, I. Application of modified carbon anodes in microbial fuel cells. Process Saf.
Environ. Prot. 2007, 85, 481–488. [CrossRef]
53. Tsai, H.-Y.; Wu, C.-C.; Lee, C.-Y.; Shih, E.P. Microbial fuel cell performance of multiwall carbon nanotubes on carbon cloth as
electrodes. J. Power Sources 2009, 194, 199–205. [CrossRef]
Processes 2023, 11, 163 19 of 19

54. Kirubaharan, C.J.; Santhakumar, K.; Senthilkumar, N.; Jang, J.-H. Nitrogen doped graphene sheets as metal free anode catalysts
for the high performance microbial fuel cells. Int. J. Hydrogen Energy 2015, 40, 13061–13070. [CrossRef]
55. Wang, Y.; Zhao, C.-e.; Sun, D.; Zhang, J.-R.; Zhu, J.-J. A graphene/poly (3, 4-ethylenedioxythiophene) hybrid as an anode for
high-performance microbial fuel cells. ChemPlusChem 2013, 78, 823. [CrossRef]
56. Christwardana, M.; Frattini, D.; Accardo, G.; Yoon, S.P.; Kwon, Y. Effects of methylene blue and methyl red mediators on
performance of yeast based microbial fuel cells adopting polyethylenimine coated carbon felt as anode. J. Power Sources 2018, 396,
1–11. [CrossRef]
57. Singh, S.; Songera, D.S. A review on microbial fuel cell using organic waste as feed. CIBTech J. Biotechnol. 2012, 2, 17–27.
58. Al-Zaqri, N.; Yaakop, A.S. Potato waste as an effective source of electron generation and bioremediation of pollutant through
benthic microbial fuel cell. Sustain. Energy Technol. Assess. 2022, 53, 102560.
59. Sasaki, D.; Sasaki, K.; Tsuge, Y.; Kondo, A. Less biomass and intracellular glutamate in anodic biofilms lead to efficient electricity
generation by microbial fuel cells. Biotechnol. Biofuels 2019, 12, 72. [CrossRef]
60. Li, M.; Zhou, M.; Tian, X.; Tan, C.; McDaniel, C.T.; Hassett, D.J.; Gu, T. Microbial fuel cell (MFC) power performance improvement
through enhanced microbial electrogenicity. Biotechnol. Adv. 2018, 36, 1316–1327. [CrossRef]
61. Idris, M.O.; Kim, H.C. Exploring the effectiveness of microbial fuel cell for the degradation of organic pollutants coupled with
bio-energy generation. Sustain. Energy Technol. Assess. 2022, 52, 102183.
62. Yaqoob, A.A.; Mohamad Ibrahim, M.N.; Umar, K.; Bhawani, S.A.; Khan, A.; Asiri, A.M.; Khan, M.R.; Azam, M.; AlAmmari, A.M.
Cellulose derived graphene/polyaniline nanocomposite anode for energy generation and bioremediation of toxic metals via
benthic microbial fuel cells. Polymers 2021, 13, 135. [CrossRef] [PubMed]
63. Yaqoob, A.A.; Idris, M.O.; Ahmad, A.; Daud, N.N.M.; Ibrahim, M.N.M. Removal of Toxic Metal Ions from Wastewater Through
Microbial Fuel Cells. In Microbial Fuel Cells for Environmental Remediation; Springer: Singapore, 2022; pp. 299–325.
64. Umar, M.F.; Rafatullah, M.; Ibrahim, M.N.M.; Ismail, N. Bioelectricity production and xylene biodegradation through double
chamber benthic microbial fuel cells fed with sugarcane waste as a substrate. J. Hazard. Mater. 2021, 419, 126469. [CrossRef]
65. Huang, L.; Chai, X.; Quan, X.; Logan, B.E.; Chen, G. Reductive dechlorination and mineralization of pentachlorophenol in
biocathode microbial fuel cells. Bioresour. Technol. 2012, 111, 167–174. [CrossRef] [PubMed]
66. Sharma, Y.; Li, B. The variation of power generation with organic substrates in single-chamber microbial fuel cells (SCMFCs).
Bioresour. Technol. 2010, 101, 1844–1850. [CrossRef] [PubMed]
67. Salvin, P.; Ondel, O.; Roos, C.; Robert, F. Energy harvest with mangrove benthic microbial fuel cells. Int. J. Energy Res. 2015, 39,
543–556. [CrossRef]
68. Fadzli, F.; Yaakop, A. Benthic microbial fuel cells: A sustainable approach for metal remediation and electricity generation from
sapodilla waste. Int. J. Environ. Sci. Technol. 2022, 14, 1–14.
69. Yaqoob, A.A.; Ibrahim, M.N.M.; Guerrero-Barajas, C. Modern trend of anodes in microbial fuel cells (MFCs): An overview.
Environ. Technol. Innov. 2021, 23, 101579. [CrossRef]
70. Serrà, A.; Bhawani, S.A.; Ibrahim, M.N.M.; Khan, A.; Alorfi, H.S.; Asiri, A.M.; Hussein, M.A.; Khan, I.; Umar, K. Utilizing
Biomass-Based Graphene Oxide–Polyaniline–Ag Electrodes in Microbial Fuel Cells to Boost Energy Generation and Heavy Metal
Removal. Polymers 2022, 14, 845.
71. Yaqoob, A.A.; Guerrero–Barajas, C.; Umar, K. Local fruit wastes driven benthic microbial fuel cell: A sustainable approach to
toxic metal removal and bioelectricity generation. Environ. Sci. Pollut. Res. 2022, 29, 32913–32928. [CrossRef]

Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual
author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to
people or property resulting from any ideas, methods, instructions or products referred to in the content.