Arabian Journal of Chemistry (2021) 14, 103415

King Saud University

Arabian Journal of Chemistry

www.ksu.edu.sa
www.sciencedirect.com

ORIGINAL ARTICLE

Bionanocomposite of Au decorated MnO2 via

in situ green synthesis route and antimicrobial
activity evaluation
M. Rahmat a, H.N. Bhatti a, A. Rehman b, H. Chaudhry c, M. Yameen d,*, M. Iqbal e,*,
S.H. Al-Mijalli f,*, N. Alwadai g, M. Fatima h,*, M. Abbas i

a
Department of Chemistry, University of Agriculture, Faisalabad, Pakistan
b
National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan
c
Environmental Science Department, Lahore College for Women University, Lahore, Pakistan
d
Department of Biochemistry, Government College University, Faisalabad, Pakistan
e
Department of Chemistry, The University of Lahore, Lahore, Pakistan
f
Biology Department, College of Science, Princess Nourah bint Abdulrahman University (PNU), Riyadh, Saudi Arabia
g
Department of Physics, College of Science, Princess Nourah bint Abdulrahman University (PNU), Riyadh 11671, Saudi Arabia
h
Department of Physics, Deanship of Educational Services, Qassim University, Buraydah, Saudi Arabia
i
Department of Biochemistry, University of Veterinary and Animal Sciences, Lahore, (Jhang Campus), Jhang 35200, Pakistan

Received 17 May 2021; accepted 30 August 2021

Available online 06 September 2021

KEYWORDS Abstract Green synthesis gaining a significant importance for the preparation of nanoparticles
Bionanocomposites; (NPs) and NPs-based biocomposites gained much attention in biological applications. In the cur-
Cinnamon extract; rent study, gold (Au) nanoparticles were prepared via green approach using cinnamon extract.
Manganese oxide nanofibers; The Au nanocomposite (NC) was prepared with MnO2 nanofiber mesh structure. The NC was
Gold nanoparticles; characterized by XRD, SEM, FT-IR, EDX, UV–visible and DLS techniques. The MnO2 nanofibers
Green approach; diameter was in 10–25 nm range, which was arranged in a mesh form and Au NPs was combined
Antimicrobial activity with nanofibers randomly. The MnO2-Au NC antimicrobial activity was measured against Escher-
ichia coli, Pseudomonas aeruginosa and Staphylococcus aureus strains. The antimicrobial activity of
MnO2-Au NC was highly promising against tested microorganisms in comparison to control (cipro-
floxacin, a standard drug). The antimicrobial activity of MnO2-Au NC was found in following
order; > S. aureus > E. coli > P. aeruginosa with the zones inhibition of 22, 18 and 15 (mn),
respectively. The MIC (minimum inhibitory concentration) values were 316, 342 and 231 (mg/mL)

* Corresponding authors.
E-mail addresses: [email protected] (M. Yameen), [email protected] (M. Iqbal), [email protected] (S.H. Al-Mijalli),
[email protected] (M. Fatima).
Peer review under responsibility of King Saud University.

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https://doi.org/10.1016/j.arabjc.2021.103415
1878-5352 Ó 2021 Published by Elsevier B.V. on behalf of King Saud University.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
2 M. Rahmat et al.

for E. coli, P. aeruginosa and S. aureus, respectively. In view of promising antimicrobial activity, the
MnO2-Au NC prepared via green approach could have potential applications in medical field and
future study can be engrossed on the biocompatibility evaluation of MnO2-Au NC using bioassays.
Ó 2021 Published by Elsevier B.V. on behalf of King Saud University. This is an open access article under
the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction The Au NPs deposition of metal oxide have been demonstrated

to promote the oxygen evolution in a reaction . Recently, Sun
The resistance of microbes against antibiotics is one of main et al also prepared MnO2 composites with good catalytic activ-
factors for the inefficacy of antibiotics against infectious ity (Sun et al., 2017) and Gorlin et al reported the deposition of
microorganisms, which drawn a significant consideration of Au NPs on MnOx by sputtering, which showed higher activity
researchers to develop new antimicrobial agents. In this versus MnOx (Gorlin et al., 2014). As mention in Table 1, a lot
regard, the nanotechnology meets this challenge and furnished of studies have been performed for Au NPs preparation by
NPs having typical properties and diverse applications in every green for different applications. However, Cinnamon extract
life sphere. The noble metals-based NPs gained much attention has not been utilized to fabricate the Au NPs. Hence, the
especially for biological applications due to their ideal therag- present study was designed to prepare the Au NPs using
Cinnamon extract and its composite with MnO2 was also
nostic aptitudes (Nazir et al., 2021; Safardoust-Hojaghan
prepared. The as-synthesized material was studied by XRD,
et al., 2021; Ghanbari and Salavati-Niasari, 2021) and Au
SEM, EDX, FTIR, UV–visible and DLS approaches. The
NPs are ideal for biomedical applications. At current, biocom-
MnO2-Au NC antimicrobial efficacy was studied against panel
posites are gaining importance in view of their promising prop-
of strains by disc diffusion method along with MIC.
erties versus NPs prepared via chemical synthesis. The
biocomposites have been employed successfully in the biomed-
ical field. The Au NPs have been fabricated by employing dif- 2. Material and methods
ferent techniques, i.e., chemical, physical and biological routes
(Karami et al., 2021; Mortazavi-Derazkola et al., 2017; 2.1. Chemicals and reagents
Salavati-Niasari et al., 2009). Among synthesis approaches,
the green route is an eco-benign and sustainable approach 2.1.1. Extract preparation
being used for the designing of nanomaterials since no haz- The chemical and reagents were acquired form Sigma-Aldrich.
ardous chemical is involved during green processing versus The Cinnamon extract was prepared and used for the fabrica-
chemically routes (Masjedi-Arani and Salavati-Niasari, 2017; tion of Au NPs. Briefly, 0.4 g of dried Cinnamon powder was
Beshkar et al., 2017; Zinatloo-Ajabshir et al., 2016; Yapaoz dispersed in 100 mL of deionized water and heating was per-
and Attar, 2019). Different bioresources have been used to fab- formed at 100 °C for 2 h. Then, the contents are filtered and
ricate the nanomaterial, which furnished the NPs of different resultant extract was kept at 4 °C for further use.
sizes and morphologies (Tavakoli et al., 2015; Ansari et al.,
2018; Bulut Kocabas et al., 2020). Plants are the rich source 2.2. MnO2-Au NC preparation
of bioactive compounds like, sugars, vitamins and polypheno-
lics that can severe as an agent for the reduction of ions to Manganese oxide nanofibrous mesh was fabricated following
atom and bioactive in the extract also facilitates the capping previously reported protocol (Tehseen et al., 2018). In a typical
and stabilization of NPs and green route is virtuous substitute synthesis scheme, MnO2 was dispersed in 100 mL deionized
of chemical synthesis approach where templates and solvents water and incubated at 80 °C with stirring at 600 rpm. Then
are used (Amer and Awwad, 2021; Awwad et al., 2020; during stirring, 5 mL of HAuCl4 (1 M) was added and incu-
Awwad et al., 2020). bated for 30 min. Now, 5 mL of Cinnamon extract was supple-
Transition metals, i.e., Ni, V, W, Mn and their oxide are mented and the content was incubated at 80 °C with stirring at
widely used for synthesis of composite materials due to their 600 rpm. The mixture was stirred for 12000 rpm for 10 min for
physical and electrochemical properties. Among Transition the separation of particle, which was washed thoroughly with
metals, the MnO2 is one of the attractive and promising mate- deionized water several time. Finally, the sample was dried at
rial for electrochemical, magnetic and catalytic application due 60 °C for 12 h and placed in a desiccator at room temperature
to its natural abundant in earth, low cost, wide potential range, till further analysis. The scheme for the synthesis of MnO2-Au
nontoxic and environmentally nature. MnO2 is also considered NC is portrayed in Fig. 1.
as promising oxidants and act as antibacterial agent, used in
sensing, photocatalysis and in pharmaceuticals applications 2.3. Characterization
(Rahmat et al., 2019; Sanda et al., 2021; Awwad et al.,
2020). The composite/doped materials facilitate the charge The XRD spectra was recorded using X’pert Pro PAN
transmission from metals to the electronegative metal, result- analytical (Germany) diffractometer using CuKa radiation
ing the formation of more stabilized oxidation state, which (k = 1.5406A), operated at 40 kV and 30 mA in 10-80° degree
have been used for different applications, i.e., Seitz el al. range. The morphology was studied by SEM (FESEM-
(Seitz et al., 2015) prepared doped MnOx for photocatalysis. JSM-7500F-JEOL, Japan). Sample was prepared by dispersing
Also, Au NPs decorated with metal showed higher catalytic powder in water (1 mg/mL) and subjected to analysis. DLS
activity for alcohol oxidation (Alhumaimess et al., 2014). experiment was performed for determination of particle size
Bionanocomposite of Au decorated MnO2 3

Table 1 Comparison of antimicrobial activity of MnO2-Au NC with different nanomaterials and NCs.
S. No NPs and NCs Tested strains ZOI (mm) References
1 MnO2 and Ag NPs E. coli, E. faecalis 5–16 (Mahlangeni et al., 2020)
2 MnO2 nanocolloids P. aeruginosa, E. coli, S. typhi 32–35 (Rahmat et al., 2019)
3 Mn NPs S. aureus, E. coli 16–24.5 (Kamran et al., 2019)
4 Zeolite\ZnO-CuO B. subtilis 18.9 (Alswat et al., 2017)
NCs
5 TiO2/ZnO NCs S. aureus 19.5 (Suo et al., 2019)
E. coli 23
6 Ag NPs P. aeruginosa 10 (Ajaypraveenkumar et al., 2015)
7 MnO2-Au NC E. coli 24 Present work
S. aureus 30
P. aeruginosa. 14

Fig. 1 Scheme adopted for the synthesis of MnO2-Au NC.

as well as surface charge of the prepared martial (Nano-ZS; ture. The Patri plates were kept at room temperature till the
Malvern Instruments). FT-IR was used to determine the func- media was solidified. With the help of forcep, disc containing
tional groups using Perkin Elmer System 2000 FT-IR spec- 500 lL MnO2-Au NC (50 mg/mL) were placed flat on the
trometer. UV–visible was performed in 200–700 nm domain medium (growth media) and Petri plates were incubated at
(CECIL (Aquarius) 7000 series). For EDX analysis Hitachi 37 °C for 24 h. The NC inhibited the bacterial growth, which
S-4800 Japan was used. was recorded with zone reader in mm. All the run was per-
formed in triplicate and data obtained was averaged. For the
2.4. Antimicrobial activity measurement of MIC, the method reported elsewhere was fol-
lowed (Sarker et al., 2007).
The antimicrobial activity was evaluated as reported elsewhere
(Rahmat et al., 2019). Antimicrobial activity was examined for 3. Results and discussion
S. aureus; P. aeruginosa and E. coli strains. The media (Nutri-
ent agar, a growth media for bacteria) was autoclaved at 3.1. XRD analysis
121 °C for the period of 15 min. The wicks paper discs
(9 mm) were sterilized. A 20 mL media (Nutrient agar) was The structural analysis was performed by XRD study and
added in Patri plates along with 20 lL of fresh bacterial cul- response thus obtained is shown in Fig. 2. The peaks recorded
4 M. Rahmat et al.

were of 220, 300, 211, 420, 521, 202 and 312 indices and
matched precisely with JCPDS card no. 44-0141. No extra
peaks were observed, which indicate the purity of prepared
material. The composite showed a peak of Au in addition to
the MnO2, which is an indication of MnO2 composite forma-
tion with Au. Also, these findings are in line with reported
studies, i.e., a hydrothermal treatment was adopted for the
fabrication of MnO2 by (Hlaing and Win, 2012) and a very
uniform MnO2 nanorods with diameters of 30–50 nm were
obtained under control of conditions (temperature and reac-
tion time). Also, PLAL has been used for the fabrication of
MnO2 NCs. The PLAL furnished high crystalline MnO2
NCs with crystallite size of 52 nm (Rahmat et al., 2019).

3.2. UV–Vis analysis

The MnO2-Au NC UV–Vis was performed in 200 to 800 nm

range and response this observed is presented in Fig. 3. In
UV–Vis absorption spectrum, two characteristic peaks were Fig. 3 UV–VIS spectrum of MnO2-Au NC.
observed at 245 nm and 490 nm, which are corelated with
MnO2 and Au, respectively. The visual appearance of ruby
3.4. FT-IR analysis
red color also reveals the formation of Au nanoparticles.
The absorption bands in 197–270 nm and 450–785 nm range
were recorded in case of MnO2, while in case of MnO2-Au FT-IR spectra of MnO2 nanofiber mesh and MnO2-Au NPs
NC, the intensity of the absorption peak in 450–785 nm range NC are shown in Fig. 5. In the spectra, a characteristic peak
reduced significantly, which is indication of interaction Au of MnO is observe at 570 and 690 (cm 1). In case of MnO2-
with MnO2 in NC formation. Au NPs NC, a band at 3321 cm 1 was observed due to OH
stretching vibration. The band observed at 1053 cm 1 was
3.3. DLS analysis assigned to the ACOA stretching. A band at 1634 cm 1 is
due to AC‚CA stretching and bending at 2127 cm 1 is due
to CAH functional group. The analysis revealed that Cinna-
The dynamic light scattering (DLS) is an analytical tool used
mon extract comprises a variety of phytochemicals, which
to measure the hydrodynamic diameter of NPs in liquid med-
are responsible for the reduction, stabilizing of Au atom
ium. It also provides information about the surface charge of
(Fig. 6). Moreover, in comparison to the MnO2 nanofiber, a
the fabricated materials. The zeta potential of the MnO2 nano-
band at 570 and 690 (cm 1) are due to Mn-O bending vibra-
fibers mesh was 26.3 mV (Fig. 4a). The negative values
tion. While for MnO2-Au NPs NC, the absorption bands are
revealed the higher stability of the NC (Liu et al., 2014). More-
shifted slightly (blue shift) to 668 and 707 (cm 1), respectively,
over, a reduction in value of zeta potential was observed in
which is due to interaction between Au NPs and MnO2 nano-
MnO2-Au, i.e., 12.2 mV (Fig. 4b), which is also an indication
fibers. These results are also in line with reported studies,
of NC formation of MnO2 and Au NPs.
where the interaction of the hydroxyl group (OAH) with Mn
atoms is documented. The peaks appeared at 665, 641, and
611 (cm 1) were corelated with MnAO bonding. With the
growth of Au NPs on the surface of MnAO, the absorption
bands shifted to 1682, 670, 645 (cm 1) revealed the successfully
incorporation of Au atoms on the surface of MnO2 nanofibers.

3.5. Surface morphology

The properties of the NPs depend on the morphology and size

of the particles. Hence, the morphology was studied by FE-
SEM and outcome is portrayed in Fig. 7a. The MnO2 formed
a nanofiber structure and the fibers diameter was in 10–25 nm
range and length was several microns. The length to diameter
ratio was found to be enough high, which reveals the fibrous
morphology. Moreover, the MnO2 forms a mesh like structure
and the pores of the meshes were in 10–500 nm range. The
fibers are highly flexible due to their extra-long length, which
makes the loops and curves in the fibers. The mesh structure
has various advantages like it has high surface area, mesh pre-
cludes the fibers from aggregation, which resultantly may offer
high activity during catalytic and photocatalytic activity. The
Fig. 2 XRD pattern of MnO2-Au NC.
Bionanocomposite of Au decorated MnO2 5

Fig. 4 (a) Zeta potential of MnO2 and (b) MnO2-Au NC.

3.7. Antimicrobial activity

The MnO2-Au NC antimicrobial activity was assessed against

three microbial strains, which was compared with standard
antibiotic ciprofloxacin. The antimicrobial activity of MnO2-
Au NPs NC was perfumed versus S. aureus, P. aeruginosa
and E. coli strains and responses are presented Fig. 9. The
ciprofloxacin was used as a standard to compare the antimi-
crobial activity of MnO2-Au NPs. The MnO2-Au NPs showed
excellent antimicrobial activity against selected microbes and
the zones of inhibition were in following order; S.
aureus > E. coli > P. aeruginosa. The zones of inhibition in
case of S. aureus, E. coli and P. aeruginosa were recorded to
be 22, 18 and 15 (mm), respectively, whereas this value was
32–36 mm in case of ciprofloxacin in case of S. aureus,
E. coli and P. aeruginosa. Previous studies also supports the
findings of present investigation that composite based are good
Fig. 5 FT-IR spectra of MnO2 NFs mesh and MnO2-Au NC. antimicrobial agents, i.e., E. coli growth was significantly
inhibited by the Ag/MnO2 NCs (Rahmat et al., 2019). Also,
Ag doped MnO2 showed promising activity against S. aureus,
average diameter of Au NPs was estimated to be 14 nm (Fig. 7- S. epidermis, B. subtilis, E. coli, S. abony and K. pneumonia
a-d), which are randomly distributed over the surface of the strains (Kunkalekar et al., 2014). Similarly, CS-MnO2 also fur-
MnO2 nanofibers. Au NPs with high electronegativity can nished auspicious antimicrobial activity against panel of
affect the physicochemical attributes of MnO2 nanofibers microbial strain (Anwar, 2018) and ternary nanocomposite
due to ideal surface-to-volume ratio (Liu et al., 2014). of MnO2 also showed auspicious activity against E. coli and
S. aureus strains (Venkatesh et al., 2017). Also, present inves-
3.6. Elemental analysis tigation revealed that the MnO2-Au NC is highly active
antimicrobial agent, which could have potential biological
The elemental composition of the MnO2-Au NC was studied application as an antimicrobial agent (Table 1). Previous stud-
by EDX analysis and response this obtained is depicted in ies showed that TiO2/ZnO NCs and ZnO NPs, the MnO2-Au
Fig. 8. The EDX analysis furnished data, which contains peaks NC showed higher antimicrobial activity. In comparison to Ag
corresponding to the elemental composition of the sample NPs along, the MnO2-Au NC also showed higher efficiency.
being analyzed. EDX spectrum of MnO2-Au NC depicts the The MIC was also studied of MnO2-Au NC against S. aur-
presence of C, O, Mn and Au elements. No peak for impurity eus, P. aeruginosa and E. coli and results are portrayed in
was observed ensuring purity of the prepared composite mate- Fig. 10. The MIC value is the considered the lowest concentra-
rial. It can be concluded that a successful decoration of Au tion of the tested compound, which inhibited the microbial
NPs was achieved on MnO2 nanofiber mesh structure. The growth (no growth observed at MIC value). The MIC values
Mn and Au proportion in the NC was 10.14 and 1.94 (%), were found to be in line with the antibacterial activity. The
respectively. The findings revealed that the MnO2-Au NC fab- MIC value was 231 mg/mL (lowest) in case of S. aureus,
ricated are in highly pure form since no additional peaks are whereas this value was 316 mg/mL for E. coli and 342 mg/mL
detected in the EDX spectrum. in case of P. aeruginosa. The standard antibiotic (ciprofloxa-
6 M. Rahmat et al.

Fig. 6 FT-IR spectra of extract used for the preparation of Au NPs.

Fig. 7 FE-SEM analysis, (a) MnO2 nanofibers and (b-d) MnO2-Au NC.

cin) showed the MIC values 105, 126 and 61 (mg/mL) against release of metabolites, metabolic functions and distribution in
E. coli, P. aeruginosa and S. aureus, respectively. The higher the cell. The NPs binds with the cell of microorganism,
antibacterial activity is due to the different interactive effect attached with the surface through active moieties and then, dif-
of MnO2-Au NC with microbial cell due to ions formation fused inside the cell and interactive with the cell different com-
and reactive oxygen species (ROS) generation, which collec- ponents. For example, if the NPs binds/interact with proteins,
tively leads to cell to death (Fig. 11). The antibacterial activity which is an important biomolecule in cell to perform different
of the NPs depends upon various factors, i.e., absorption rate, functions, i.e., it takes part in the formation of cell wall, ribo-
Bionanocomposite of Au decorated MnO2 7

Fig. 8 (a) EDX analysis of MnO2 nanofibers and (b) MnO2-Au NC.

Fig. 10 Minimum inhibitory concentration (MIC, mg/mL)

Fig. 9 Antimicrobial activity (zone of inhibition) of MnO2-Au
MnO2-Au NC.
NC.

mation of cell components (wall, membrane, ribosome and

somes, cell membrane and nucleic acids and if protein synthe-
nucleic acids) and if protein synthesis is inhibited inside the cell
sis is inhibited inside the cell and them, all activities related to
and them, all activities related to protein are stopped and
protein are stopped and resultantly, cell death occur. The over-
resultantly, cell death occur. The overall proposed antimicro-
all proposed antimicrobial mechanism is explained in Fig. 10.
bial mechanism is explained in Fig. 10. The binds with cell wall
The binds with cell wall of microbe, penetrates into cell mem-
of microbe, penetrates into cell membrane and cause mem-
brane and cause membrane destruction and disrupt (proteins,
brane destruction and disrupt (proteins, enzymes, and
enzymes, and DNA). The NPs produce ROS inside the cell and
DNA). The NPs produce ROS inside the cell and these ROS
these ROS induce oxidative stress and resultantly, metabolic
induce oxidative stress and resultantly, metabolic pathways
pathways are disturbed and cell occurred.
are disturbed and cell occurred. These ROS include hydroxyl
The antibacterial activity of the NPs depends upon various
radicals (OH), and superoxide ions O2  These free radicals
factors, i.e., absorption rate, release of metabolites, metabolic
interact with biomolecules, which disintegrate the plasma
functions and distribution in the cell. The NPs binds with the
membrane and cause lipid oxidation. The hydrogen peroxide
cell of microorganism, attached with the surface through active
produced cause toxicity to the cell and ultimately, leads to
moieties and then, diffused inside the cell and interactive with
the cell death (Sharma et al., 2021). The NPs and NC prepared
the cell different components. For example, if the NPs binds/
promising antibacterial activity versus related nanomaterials
interact with proteins, which is an important biomolecule in
prepared chemical methods. Hence, the adoption of green
cell to perform different functions, i.e., it takes part in the for-
route is more feasible to prepare the NPs (Awwad et al.,
8 M. Rahmat et al.

Fig. 11 Antimicrobial activity mechanism of MnO2-Au NC.

2020; Awwad et al., 2020; Awwad and Amer, 2020; Al Banna which need further studied for the evaluation of biocompatibil-
et al., 2020; Igwe and Nwamezie, 2018; Remya et al., 2017) and ity of MnO2-Au NC.
their composites for biomedical applications.
Declaration of Competing Interest
4. Conclusions
The authors declare that they have no known competing
The Au NPs was prepared by green route using cinnamon financial interests or personal relationships that could have
extract and nanocomposite of Au was prepared with MnO2 appeared to influence the work reported in this paper.
nanofibrous mesh. The characteristics of MnO2-Au NC was
studied by SEM, XRD, FT-IR, EDX, UV–visible and DLS Acknowledgement
techniques. The MnO2 nanofibers diameter was in 10–25 mm
range. The length to diameter ratio was found to be enough This research was funded by the Deanship of Scientific
high, which reveals the fibrous morphology. The MnO2 forms Research at Princess Nourah bint Abdulrahman University
a mesh like structure and the pores of the meshes were in 10– through the Fast-track Research Funding Program.
500 nm range and Au atom was distributed randomly on the
surface of nanofibers. The MnO2-Au NC showed promising Reference
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