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Graphene Oxide-ZnO Nanorods for Efficient Dye Degradation, Antibacterial

and In-silico Analysis

Article  in  Applied Nanoscience · January 2022

DOI: 10.1007/s13204-021-02251-2

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Applied Nanoscience
https://doi.org/10.1007/s13204-021-02251-2

ORIGINAL ARTICLE

Graphene oxide‑ZnO nanorods for efficient dye degradation,

antibacterial and in‑silico analysis
Saira Shaheen1 · Azhar Iqbal1 · Muhammad Ikram2   · Muhammad Imran3 · Sadia Naz4 · Anwar Ul‑Hamid5 ·
Anum Shahzadi6 · Walid Nabgan7 · Junaid Haider4 · Ali Haider8

Received: 31 July 2021 / Accepted: 13 November 2021

© King Abdulaziz City for Science and Technology 2021

Abstract
Numerous concentrations of graphene oxide (GO)-doped into ZnO nanostructures were synthesized with co-precipitations
method. The characterization of the designed composite was carried out using a number of techniques, and their photocata-
lytic ability was also evaluated. The parameters such as plane orientation, interplanar distance as confirmed using XRD
were consistent with HRTEM results. The PL analysis revealed a blueshift in the energies associated with NBEs, which
verified the impact of dopant quenching. The findings indicate that ZnO was loaded uniformly on the GO surface due to
an efficient interface coupling. Due to such interface coupling between ZnO and GO, electrons can be passed directly from
the ZnO (VB) to the GO. The facile nanocomposites (NCs) exhibited excellent photoactivity and are considered to offer a
new path for designing next-generation graphene-based semiconductor composites. Furthermore, as-synthesized samples
exhibited promising bactericidal potential towards gram-positive (G + ve) and gram-negative (G − ve) bacteria in S. aures
and E. coli media, respectively for bacterium. Furthermore, molecular docking studies was employed to unveil the mystery
behind bactericidal activity of GO-ZnO NRs and suggested inhibition of β-lactamase and DNA gyrase as mechanism behind
these in vitro findings.

Keywords  Graphene · ZnO · Nanorods · Antimicrobial activity · HRTEM · Molecular docking

Introduction woodworking, leather, paper, inks, paints, and fabric tex-

tiles have been discharging a wide variety of synthetic dyes
Water is the basis of life and a necessary component for and pigments into the environment (Wang et al. 2018; Khan
the survival of all living creatures. From the 71% of water et al. 2019a, b). According to research, the leather and tex-
present on the surface of the earth, only 0.03% water is tile industries are significant contributors to the produc-
useable for drinking purposes. Since the last four decades, tion of large quantities of textile effluent, which accounts
industries such as medicine, pharmaceuticals, food, fibers, for approx. 20% water contamination, as assessed by the

* Muhammad Ikram Catalysts, Beijing University of Chemical Technology,

[email protected] Beijing 100029, China
4
* Anwar Ul‑Hamid Tianjin Institute of Industrial Biotechnology, Chinese
[email protected] Academy of Sciences, Tianjin 300308, China
5
* Walid Nabgan Core Research Facilities, King Fahd University of Petroleum
[email protected] and Minerals, Dhahran 31261, Saudi Arabia
6
1 Faculty of Pharmacy, University of the Lahore, Lahore,
Department of Physics, School of Science, University
Pakistan
of Management and Technology, Lahore 54000, Pakistan
7
2 School of Chemical and Energy Engineering, Faculty
Solar Cell Applications Research Lab, Government College
of Engineering, Universiti Teknologi Malaysia,
University, Lahore 54000, Pakistan
81310 Skudai, Johor, Malaysia
3
State Key Laboratory of Chemical Resource Engineering, 8
Department of Veterinary Medicine, University of Veterinary
Beijing Advanced Innovation Centre for Soft Matter Science
and Animal Sciences, Lahore 54000, Punjab, Pakistan
and Engineering, Beijing Engineering Center for Hierarchical

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 Applied Nanoscience

World Bank (Kant 2012; Anliker 1979). With the rise of water and air (Nazari 2019). Catalytic performance of ZnO
industrialization, the issue of water pollution has become an can be very low because of recombination of excitons and
everyday concern. Numerous methods are currently used to resulting low photo reacting sequence. As a result, vari-
purify wastewater, including membrane filtration, adsorp- ous approaches are used to improve the absorption range
tion, reverse osmosis, distillation, electrochemistry, phy- of ZnO or to inhibit electron–hole recombination (Mad-
toremediation, and photocatalysis (Siripireddy and Mandal husudhana Reddy et al. 2017). These techniques include
2017). In the past, many scientists have worked successfully structural modeling, noble metal doping, ion doping, and
to produce new semiconductor photocatalysts. Numerous semiconductor coupling. On the other hand, ZnO has certain
low-cost, efficient, and advanced photocatalytic materials drawbacks, including uneven dispersion, extreme resistiv-
were developed and used in photochemical sewerage treat- ity, and shallow mixture of photoengraved electron–hole
ment. Various metal oxides were extensively used for this pairs (Han et al. 2014). Additionally, ZnO is activated only
purpose including ZnO (Hong et al. 2018; Pruna et al. 2018; by UV irradiation, which has low light quantum efficiency.
Pradhan et al. 2012), ­TiO2 (Nagamine and Inohara 2018), Numerous studies have been performed to test the photo-
sulfides, and oxynitrides (Soignard et al. 2005). catalytic efficiency in the NCs (Kumar et al. 2021). ZnO
Graphene is a single-layer carbon material, with remark- can be synthesized using various methods such as sol–gel
able electrical and mechanical properties. It possesses lay- (Haan et al. 2018), precipitation (Moharram et al. 2014; Lin
ered structure, extraordinary electro-transport, good elec- et al. 2020), solvothermal (Atchudan et al. 2016) and hydro-
trical properties and high photocatalytic activity. Graphene thermal (Kavitha et al. 2015). Mostly ZnO and GO can be
and its derivatives play a significant role in semiconductor used a photocatalyst. Recently, these two nanocomposites
industry owing to the absence of a large band gap (Mututu (GO-ZnO) attracted scientists’ attention due to their high
et al. 2019). It also combines with inorganic nanoparticles adsorption capacity and the presence of graphene oxide
such as ­TiO2, ­SnO2 and ZnO to display useful properties with a narrow band. This results in active catalysis under
such as high efficiency of solar and fuel cells and photocata- visible light (Hashim et al. 2016). It is also observed that
lytic reduction. Carbon or carbonaceous materials are highly the photocatalytic activity of GO-ZnO NCs is raised ZnO
desirable due to their special properties, which include a compared to ZnO in methylene orange (Toporovska et al.
porous structure, satisfactory electronic properties, and 2020), methylene blue (Zarrabi et al. 2018), methylene violet
adsorptive capability. Activated carbon, carbon nanotubes, (Punithavathy et al. 2017) and bromophenol blue (Moorthy
and graphene are all included in this category (Zhang et al. et al. 2017). This occurs due to electron recombination with
2015). graphene oxide. Further, it exhibits very little transfer resist-
From literature search, it can be concluded that ZnO and ance, high diffusion and high electron transfer rate on ZnO.
graphene represent one of the best combination for improved It plays an important part in GO-ZnO material for wastewa-
photocatalytic efficiency. This is due to the graphene’s con- ter treatment.
jugated structure, moreover, it provides separation of charges Taking into account the aforementioned attributes of
during photocatalysis method (Ahmad et al. 2015). Corre- GO and ZnO, the aim was to combine these two to develop
spondingly, graphene oxide (GO) contains hydroxyl and car- facile, safer and inexpensive GO-doped ZnO NRs compos-
boxyl groups that are dispersible in solvents, allowing for the ites with reduced toxicity and more stability than employed
fabrication of GO-based nanohybrids composites (Raizada separately for two specific practical applications. Synthesis
et al. 2019). Photogenerated electrons traveling toward the was undertaken followed by antimicrobial potential evalu-
surface interact with the layer of hybrid GO, thus increasing ation against Escherichia coli (E. coli) and Staphylococcus
photocatalytic ability. This reduces the chances of recom- aureus (S. aureus). Significant catalysis application against
bination of photogenerated excitons pairs, thus as a result methylene blue, a common industrial dye, was also achieved
electrons and holes (excitons) are separated (Bu et al. 2013). successfully in the presence ­NaBH4. In addition, in silico
Specifically, for the degradation of organic dyes, GO con- molecular docking studies were carried out against selected
tributes significantly to the photodegradation productivity of enzyme targets namely β-lactamase and DNA gyrase from
photocatalysts, owing to heavy adsorption between GO and both E. coli and S. aureus.
a variety of organic dyes (Zhang et al. 2012).
A large number of metal oxides are employed as photo-
catalysts to purify water. T ­ iO2, ZnO, F­ e2O3, ­ZrO2, ­V2O5, Experimental part
­Nb2O5, and ­WO3 are commonly used (Raizada et al. 2017a,
b). Among these, ZnO is an important photocatalyst because Materials
of its high stability and bandgap energy (Eg) of 3.3 eV. It
is non-toxic and exhibits strong photocatalytic activity that Zinc nitrate tetra hydrate-Zn(NO3)2∙4H2O, 99%, sodium
enables it to remove different waste material from both the borohydride ­(NaBH4, 98%) and sodium hydroxide (NaOH,

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98%) of analytical grade were acquired from Sigma-Aldrich, 90 °C. As a result, ultra-fine powder of GO-doped ZnO
Germany. Sulfuric acid ­(H2SO4) was purchased from Analar nanocomposites was obtained as shown in Fig. 1a, b.
(USA).
Materials characterization
Synthesis of (GO‑ZnO)
X-ray diffraction with a powder diffractometer (PAN Analyt-
Chemical precipitation was used to synthesize ZnO ical X' pert PRO model) and monochromatic Cu-Kα radia-
nanorods. Initially, Zn ­(NO3)2∙4H2O (0.1 M) was mixed in tion (λ = 1.5418 Å) at a scan rate of 5°/min for 2θ (5°–80°)
deionized water (DI water) under continuous stirring to form was utilized to analyze structural and crystalline character-
a solution. NaOH was then added drop by drop to retain the istics of synthesized powder. On the Excalibur 3100 spec-
solution's pH at 12. After 2 h at 90 °C, precipitates were trometer, Fourier Transform Infrared (FTIR) spectroscopy
observed. For the synthesis of GO from refined graphite was employed within a spectral range of 4000–400 ­cm−1.
powder, a modified Hummers method was used. Subse- FE-SEM (JEOL JSM-6460LV) and HR-TEM (JEOL JEM
quently, various concentrations (2, 4 and 6%) of prepared 2100F) were utilized to analyze interlayer, spacing morphol-
GO were dispersed ultrasonically in DI water for 2 h. Sub- ogy and particle size. Using a Genesys 10S UV–vis spec-
sequently precipitated solution of GO was transferred to the trophotometer in the range of 180–900 nm, optical proper-
above-mentioned solution of ZnO (0.1 M). It was then cen- ties were investigated. Spectra of photoluminescence (PL)
trifuged at 4000 rpm for 20 min and washed with DI water. were recorded with a spectrofluorometer for as-prepared and
Eventually, oven was used to dry the product overnight at doped samples (JASCO, FP-8300).

Fig. 1  Schematic representation of the preparation of a graphene oxide b GO/ZnO nanorods

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Results and discussion photocatalysis and with an ultrasonic bath called sonocataly-

sis independently and simultaneously (sonophotocatalysis),
Catalysis as shown in Fig. 3. The sonocatalytic and sonophotocatalytic
degradation rates were determined using ultrasonic baths
The mechanism of dye degradation is shown in Fig.  2. with a rectangular shape (XUB6; Grant Co., Ltd., Eng-
­BH4− delivers electron to MBCF through GO/ZnO NRs fol- land) operating at a constant frequency of 35 kHz at 200 W
lowing dissociation of ­NaBH4. The most significant function applied power. The glass beaker was positioned at the center
in dye degradation is played by NRs, which absorbs and sub- of ultrasonic bath.
sequently transmits the electron. Reduction in redox poten- A Genesys 10S UV–vis spectrophotometer was used
tial actually happens with the rise in Fermi potential, which to regulate photocatalysis and sonophotocatalysis of GO-
probably is controlled by the size of NRs. Particle size plays ZnO NRs under a mercury lamp (400 W) that was used to
a very critical role in the interfacial potential barrier, for produce visible light with a λ of 400–700 nm. 10 mg of
small-sized particles the barrier is small while the surface
area is large, and vice versa. We explored the catalytic activ-
ity (CA) of GO-ZnO product by combining 3.0 ml MB solu-
tion with 800 μl freshly made ­NaBH4 solution. Following
that, 400 μl of diluted synthesized ZnO sample was applied
to the above solution. The blue color of MB was removed,
indicating that the dye had degraded. At different intervals,
the absorption spectroscopy was performed using a UV–Vis
spectrophotometer set to a wavelength range of 200–800 nm.
The obtained results of MB degradation are shown in Fig.
S1 (see supplementary material).

Photocatalytic, sonocatalytic
and sonophotocatalytic activity

The RhB degradation (dye pollutant) in the existence

of GO-ZnO was studied through visible light known as
Fig. 3  Photocatalysis mechanism of prepared samples

Fig. 2  Schematic representa-
tion of catalysis mechanics of
(GO-ZnO)

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prepared nanocatalyst was submerged in MB dye solution MSA plates of 6-mm wells created using a sanitized well
of 60 mL and it was stirred rigorously for 10 min, to get the borer. Antibacterial strength was determined using a Ver-
dye and catalyst into equilibrium. A set duration of visible nier scale after plates were incubated for 12 h at 37 °C. The
light was used to irradiate the beaker. Throughout the expo- microbicidal effect of GO-doped ZnO NRs was determined
sure period, 4-ml samples were obtained for UV analysis statistically using one-way variance analysis (ANOVA)
at regular time intervals. The resulting dye concentration (Ahmad et al. 2021).
was determined by measuring the shift in the λmax = 665 nm
of MB. The % degradation was measure with the equation Molecular docking studies
(Zarrabi et al. 2018).
c0 − ct Disrupting the cell wall biosynthesis pathway has been
%Degradation = × 100 (1) proposed as an appealing target for antibiotic exploration
c0
and as a potential mechanism underlying the bactericidal
where C0 MB solution initial concentration and Ct final function of various nanostructures. Enzyme belonging to
concentration after a fixed irradiation time at λ = 665 nm. peptidoglycan synthesis have huge significance for the dis-
Photocatalytic degradation of organic pollutants, semi- covery of new antibacterial agents as their inhibition cause
conductive optical stimulation occurred at the catalyst sur- rapture of cell wall and ultimately bacterial death (Lim and
face, accompanied by the creation of electron–hole pairs. Strynadka 2002; Buxeraud and Faure 2016). Similarly,
The catalyst's high oxidative ability enables the direct oxida- enzymes belonging to nucleic acid biosynthesis especially,
tion of pollutants (insecticide). Besides this, when the hole DNA gyrase has huge significance for antibiotics discovery
reacts with hydroxyl radicals (–OH) resulting from water (Mullis et al. 2019). Here, we selected β-lactamase and DNA
breakdown, highly reactive radicals are formed. These gyrase enzyme from both E. coli and S. aureus to evaluate
radicals are strong nonselective oxidants that speed up the inhibitory potential of GO-ZnO NRs against them.
oxidation of organic pollutants. The oxygen molecules are The 3D-crystal structures were obtained from RCSB PDB
converted to anion peroxides by the electrons in the conduc- (www.​rcsb.​org) with PDB code: 4KZ9 (Res: 1.72 Å) (Bare-
tion band (CB) of the catalyst surface. In organic pollutants, lier et al. 2014) and 5MMN (Resolution: 1.90 Å) (Panchaud
these radicals form organic peroxides or hydrogen peroxide. et al. 2017) for β-lactamase and DNA gyrase from E. coli.
Additionally, electrons in the CB are responsible for the for- Similarly, PDB ID: 1MWU (Resolution: 2.60 Å) (Lim and
mation of –OH, (Atchudan et al. 2016) which is the primary Strynadka 2002) and 4URO (Resolution: 2.59 Å) (Lu et al.
agent in organic compound mineralization (Fig. 3). Effects 2014) for β-lactamase and DNA gyrase, respectively from
of photocatalytic, sonocatalytic and sonophotocatalytic pro- S. aureus.
cesses are shown in Figs. S2–S4 (Supplementary file). The ICM Molsoft v3.8-4a or above software was engaged
for molecular docking predictions (Abagyan and Totrov
1994). The protein structures were prepared using ICM Mol-
Pathogen isolation and characterization soft that involve stepwise process, i.e., water molecules and
native ligand’s removal, adding polar hydrogen atoms, and
Ovine mastitic milk samples were procured from various optimization of protein structure using default parameters.
farms in Punjab, Pakistan and analyzed using the Surf Field Monomer unit structure of GO-ZnO NRs was built using
Mastitis Test (SFMT). Initially, materials were cultured on ligedit tool of ICM. Finally, binding pocket was specified
5% blood agar and then on specified source (MacConkey within 10 Å region of co-crystallized ligand and ten docked
agar (MCA) and mannitol salt agar (MSA) for the purpose of conformations were produced (each case). Binding interac-
sorting selective Staphylococcus aureus (SA) and Escheri- tion analysis and 3D visualization of top docked conforma-
chia coli (EC). Gram's staining, catalase, and coagulase tests tions were carried out using Discovery studio visualizer and
were used to perform taxonomic and biochemical recogni- Pymol software.
tion (Ikram et al. 2021). The XRD study of GO-doped ZnO samples revealed that
the characteristic peaks at 2θ° = 31.7°(100), 34.4°(002),
Antibacterial evaluation 36.2°(101), 47.5°(102), 56.7°(110), 63.0°(103), and
68.1°(112) observed were well matched with the literature
The well diffusion method was devised for in-vitro antibac- (Moorthy et al. 2017; Prema et al. 2019). This indicates the
terial validation of GO-doped ZnO NRs with swabbing 0.5 existence of ZnO with hexagonal wurtzite phase (JCPDS
Macfarland of distinct gram –ve and + ve segregates. In ref- No. 361451). Diffraction peaks intensity enhanced upon
erence to DIW (50 μl) and Ciprofloxacin (5 μg/50 μl), spe- GO doping into ZnO (Fig. 4a). XRD patterns suggest the
cific GO-doped ZnO NRs ratios of 500 and 1000 μg/50 μl crystalline nature of formed NRs. Observed intensity varia-
were placed as a low and high dose onto sanitized MA and tions seemed to possess no periodicity correspondence with

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Fig. 4  a XRD pattern b FTIR

spectra of ZnO, GO-ZnO (2%,
4% and 6%)

dopant concentration. However, the size of crystallite varied respectively. When the EA is in the ZnO, the excited elec-
with different concentrations of dopant while carrying vari- trons migrate through π electrons to the GO layer, emptying
ous capping and reducing agents. Crystallite size of samples the CB. As a result, the Fermi energy level is lowered and
was measure from the XRD patterns with Scherrer formula. bent toward the valence band (VB), resulting in a decrease
Crystallite size of ZnO was measured at 28 nm estimated in the Eg. In short, the narrowing of Eg in the GO-ZnO com-
using Scherrer formula. With increasing amount of GO (2, posite is due to the transition of photo-excited electrons from
4 and 6%), respectively, the measured crystallite size was the CB to GO surface through carboxyl groups on the GO-
reduced gradually down to an average of (22.5, 19.6, and ZnO interface, which offers a shorter electron transport route
16 nm), respectively. to the GO. In Fig. 5c, PL spectra of doped and undoped
Figure 4b illustrates FT-IR spectra of ZnO and GO-doped ZnO at room temperature were recorded using wavelength
NR’s. The peak intensities of the O–H and epoxy group were of 250 nm. A blueshift was observed upon GO doping into
significantly reduced in the spectra of GO-ZnO NRs, proba- ZnO NRs. PL graphs evaluate the recombination capacity
bly as a result of the interaction with GO and ZnO NRs. The of electron–hole pair after being produced by photon in pre-
above finding indicates that ZnO NRs bonded successfully pared samples. A less electron–hole recombination rate is
to the GO sheets’ surface. The peaks at 1365–1352 ­cm−1 associated with low intensity of PL. As a result, lifetime of
in the spectrum of ZnO and GO-ZnO correspond to func- charge carrier can become promisingly longer.
tional groups C = O (amide), aromatic C = C, and stretching Figure 5c illustrates the PL spectra of synthesized prod-
vibration of C–OH groups, respectively (Suresh and Yusoff ucts. For hybrid materials, a quenching emission of ZnO
2020). GO present in the material showed peak at 870 nm PL is observed. Additionally, when GO was introduced, the
(Durmus et al. 2019). GO-ZnO showed peaks at < 500 ­cm−1 intense emission of ZnO at 432 nm accompanied blueshift
(440, 458 ­cm−1) that corresponded to the vibrations of Zn–O to 405 nm and intensity decreased in comparison to the other
bonds (Nagaraj et al. 2020; Trinh et al. 2018) and the peak emission features (Chanda et al. 2020). In GO-ZnO, the near
at ~ 412 nm was assigned to ZnO stretching vibrations (Dur- band edge (NBE) of ZnO (432 nm) shifted to a shorter wave-
mus et al. 2019). length (405 nm). In contrast to pure ZnO, GO-ZnO exhib-
Figure 5a illustrate absorbance spectra of prepared NRs. ited reduced UV and blue emission and suppressed defect
The observed absorption peaks of undoped and GO-doped level emission. The reduction in the PL intensity of GO-ZnO
ZnO were found in the UV region and intensity increased implied that there were fewer excitons recombination, which
upon doping. The band gap energy (Eg) of prepared sam- is favorable for photocatalysts. The electron transfer from
ples was calculated using Tauc’ plot. Extracted values of the CB of ZnO (a strong EDGO-ZnO) to the VB of GO was
Eg from absorption spectra of ZnO and GO (2%, 4% and ascribed as the cause of the quenching (EA). This charge
6%) doped ZnO were 3.07, 3.15, 3.24 and 3.32 eV, respec- transfer, which is dependent on appropriate interactions at
tively (Fig. 5b) (Afzal et al. 2019; Alamdari et al. 2019). the interface with ZnO and GO, reduces electron–hole pair
This reduction in Eg could be the result of partial absorp- recombination. As a result, the inclusion of charge carriers
tion of light by heterogeneous bands of Zn–O–C and car- in GO-ZnO hybrids is predicted to improve ZnO’s photo-
bon structure in the samples. Chemical reaction between catalytic efficiency (Chanda et al. 2020; Víctor-Román et al.
ZnO and GO, or a joint electronic structure between them, 2020).
results in a shrinking Eg. In GO-ZnO NRs, ZnO and GO The atomic and weight percentages of different elements
behave as electron donor (ED) and electron acceptor (EA), contained in GO-ZnO NRs were measured using EDS

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Fig. 5  a UV vis. spectra of

NRs, b band gap energy and c
PL spectra of prepared samples

Fig. 6  EDS Spectrum of a ZnO, b GO/ZnO (2%), c (4%) and d (6%) NRs

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(Fig. 6a–d). Zn and O peaks verified the formation of ZnO, concentrations. Ciprofloxacin had an inhibition zone of 7.15
whereas GO signals in doped samples indicate effective inte- and 9.10 mm against S. aureus and E. coli, accordingly, in
gration of graphene atoms into the lattice. Additional peaks comparison to DI water (0 mm). At both dosages, doped
of sodium were seen in all samples since NaOH was used to GO (2, 4 and 6 wt%) showed substantial antibacterial action
maintain the pH of solutions. towards G + ve compared to G − ve.
Figure 7a–d displays HR-TEM images of ZnO, GO-ZnO The mechanism of antimicrobial reaction is shown in
(2%, 4% and 6%, respectively) that confirmed the forma- Fig. 8 for synthesized product. Because the antibacterial
tion ZnO and GO-doped NRs. To study further the crystal potency of nanoparticles is scale-dependent, the oxidative
structure and microstructural differences in ZnO NRs doped stress of developed nanomaterials is scale and concentration-
with graphene oxide, HR-TEM was employed at 100–10 nm dependent (Ahmed et al. 2019). When bacteria come into
scale. HR-TEM image disclosed high degree of agglomer- contact with nanoscale structures, reactive oxygen species
ation among NRs, which are 50 nm in size, as shown in are produced, which are fatal for cells (Ahmed et al. 2018;
Fig. 7. HR-TEM rendered lattice fringes were measured at Haider et al. 2019). Oxygen reactive species (ROS) sur-
0.28, 0.26, 0.25 and 0.23 nm for pure ZnO, GO-ZnO 2%, 4% round the bacterium's outer membrane, and bacteria die as a
and 6%, which ascertained the formation of highly crystal- result of protrusion and bulging of intracellular components
line nanostructures. (Ikram et al. 2020). Micro-pathogens also wreak havoc when
The in-vitro bactericidal potency of undoped and GO- cations form strong interactions with the negative parts of
doped ZnO NRs was determined by calculating inhibition microbial cells. Cations impair microbial ribosome func-
zones (mm) against E. coli and S. aureus with agar well dif- tions, resulting in their collapse through enzymatic degrada-
fusion technique, as shown in Table 1. For GO-ZnO NRs, tion (Altaf et al. 2020). Two responses were recognized as
the enormous inhibition domains of G + ve were verified possible for the bactericidal mechanism of nanostructures:
to be (1.85–4.7 mm) and (3.25–5.15 mm) and similarly, one entails strong interaction between cations ­Zn2+ and
(1.30–2.45  mm) and (3.45–5.75  mm) for G –ve at both microbial, resulting in the formation of negativized sections

Fig. 7  HR-TEM and interlayer

d-spacing images of ZnO and
GO-ZnO

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Table 1  Antimicrobial activity Sample S. aureus E. coli

of GO-ZnO with different
concentrations of extract Inhibition zone (mm) Inhibition zone (mm)
0.5 mg/50 μl 1.0 mg/50 μl 0.5 mg/50 μl 1.0 mg/50 μl

Reference (ZnO) 2.2 5.8 1.05 2.08

2% 1.85 3.25 1.30 3.45
4% 3.25 4.4 2.15 4.10
6% 4.7 5.15 2.45 5.75
Ciprofloxacin 7.15 7.15 9.10 9.10
DI water 0 0 0 0

Measurements of inhibition zones against G –ve

Inhibition values (mm) for G + ve

Fig. 8  Antimicrobial mecha-
nism of synthesized GO-ZnO
nanorods

and eventual collapse; the other includes electronic excita- coli. The binding interaction pattern with key amino acid
tion of the ZnO valance band surface via irradiation. Moreo- residues are depicted in Fig. 9a, b representing H-bonding
ver, the electrical O ­ 2− radicals, leading
­ 2 reaction produces O with His186 (1.8 Å), Asp229 (2.2 Å), Glu228 (2.0 Å) and
to the formation of H ­ 2O2. The subsequent O ­ 2− species are Thr226 (2.0 Å). A similar trend was observed for GO-
required for the destruction of lipid or protein molecules on ZnO NRs binding inside β-lactamase S. aureus enzyme’s
the exterior cell membrane of microbes (Haider et al. 2020; active pocket with overall binding energy −8.759 kcal/
Rajan et al. 2017). mol. Key interactions observed were H-bond with Thr600
Molecular docking predictions being an effective tech- (2.2 Å), Ser403 (2.8 and 2.7 Å) and Asn464 (2.4 Å) along-
nique to unravel the mystery behind various biological activ- side metal–contact interaction with Tyr519 as shown in
ities has attracted significant attention in the last couple of Fig. 9c, d.
decades. The significance of cell wall synthesis (i.e., pepti- The molecular docking predictions of GO-ZnO NRs
doglycan synthesis) and nucleic acid biosynthetic pathway against DNA gyrase of E. coli showed H-bonds with
for antibiotic discovery is well documented (Mullis et al. important amino acid residues such as Asp73 (2.2  Å),
2019; Higgins et al. 2005; Shaikh et al. 2015). Although Thr165 (1.7 Å) and Gly77 (2.4 Å) having total binding
antibacterial activity of various nanostructures has been score of −6.774 kcal/mol (given in Fig. 10a, b). Similarly,
reported in the last few years, the exact mechanism of their binding pattern of GO-ZnO NRs with DNA gyrase of S.
activities need to be explored further (Bury and Wood 1999; aureus are depicted in Fig. 10c, d having a binding score
Arsalan and Younus 2018). of −7.169 kcal/mol showing H-bond with Gly185 (2.3 Å),
The GO-ZnO NPs showed good binding energy Thr173 (1.8 Å) and Asp81 (2.4 and 2.8 Å).
(−7.881 kcal/mol) inside binding pocket of β-lactamase E.

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Fig. 9  Binding interaction pat-

tern with active site residues of
β-lactamase enzyme from (a),
(b). E. coli, (c), (d) S. aureus 

Fig. 10  Binding interaction pat-

tern with active site residues of
DNA gyrase enzyme from a, b
E. coli, c, d S. aureus 

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Binding tendency of GO-ZnO NRs revealed through Ahmad SOA, Ikram M, Imran M, Naz S, Ul-Hamid A, Haider A,
molecular docking predicted these NPs as potential inhibi- Shahzadi A, Haider J (2021) Novel prism shaped C ­ 3N4-doped
Fe@Co 3O 4 nanocomposites and their dye degradation and
tor of β-lactamase as well as DNA gyrase enzyme that are bactericidal potential with molecular docking study. RSC Adv
suggested to be further confirmed by in vitro enzyme inhibi- 11(38):23330–23344
tion techniques. Ahmed B, Hashmi A, Khan MS, Musarrat J (2018) ROS mediated
destruction of cell membrane, growth and biofilms of human
bacterial pathogens by stable metallic AgNPs functionalized
from bell pepper extract and quercetin. Adv Powder Technol
Conclusions 29(7):1601–1616
Ahmed B, Solanki B, Zaidi A, Khan MS, Musarrat J (2019) Bacterial
toxicity of biomimetic green zinc oxide nanoantibiotic: insights
One-pot precipitation technique was followed effectively into ZnONP uptake and nanocolloid–bacteria interface. Toxicol
to synthesize ZnO and GO-ZnO NRs. We investigated the Res (camb) 8:246–261. https://​doi.​org/​10.​1039/​C8TX0​0267C
phase structure, morphologies, elemental composition, and Alamdari S et al (2019) Preparation and characterization of GO-ZnO
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and HRTEM analyses verified the samples' crystalline com- 041
position. The plane orientation and interplanar distance Altaf S, Haider A, Naz S, Ul-Hamid A, Haider J, Imran M, Shahzadi
determined by XRD was corroborated with HRTEM. The A, Naz M, Ajaz H, Ikram M (2020) Comparative study of sele-
nides and tellurides of transition metals (Nb and Ta) with respect
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used as a highly effective photocatalyst for MB dye degra- tion of enzymatic activity via nanoparticles. Int J Biol Macromol
dation and has bactericidal behavior against gram-positive 118:1833–1847. https://​doi.​org/​10.​1016/j.​ijbio​mac.​2018.​07.​030
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tary material available at https://d​ oi.o​ rg/1​ 0.1​ 007/s​ 13204-0​ 21-0​ 2251-2. Biol 9(7):1528–1535. https://​doi.​org/​10.​1021/​cb500​1636
Bu Y, Chen Z, Li W, Hou B (2013) Highly efficient photocatalytic
Acknowledgements  Authors are thankful to HEC, Pakistan through performance of graphene-ZnO quasi-shell-core composite mate-
project 21-1669/SRGP/R&D/HEC/2017. We are extremely apprecia- rial. ACS Appl Mater Interfaces 5(23):12361–12368. https://​doi.​
tive of the support offered by the Core Research Facilities at KFUPM org/​10.​1021/​am403​149g
in Dhahran, Saudi Arabia. Bury NR, Wood CM (1999) Mechanism of branchial apical silver
uptake by rainbow trout is via the proton-coupled Na+ channel.
Declarations  Am J Physiol Regul Integr Comp Physiol 277(5):R1385–R1391.
https://​doi.​org/​10.​1152/​ajpre​gu.​1999.​277.5.​r1385
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Conflict of interest  Authors confirm no conflict of interest. 55(558S):1–5. https://​doi.​org/​10.​1016/j.​actpha.​2016.​06.​001
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