Green Synthesis of Coppercopper Oxide Nanoparticle

GREEN CHEMISTRY LETTERS AND REVIEWS
2022, VOL. 15, NO. 1, 187–215

https://doi.org/10.1080/17518253.2022.2025916
REVIEW
Green synthesis of copper/copper oxide nanoparticles and their applications:

a review
Nilanjan Chakraborty a, Jishnu Banerjee b, Pallab Chakraborty c, Anuron Banerjee a
,
Sumeddha Chanda d, Kasturi Ray d, Krishnendu Acharya e,f and Joy Sarkar b
a
Department of Botany, Scottish Church College, Kolkata, India; bDepartment of Botany, Dinabandhu Andrews College, Kolkata, India;
c
Department of Botany, Acharya Prafulla Chandra College, Kolkata, India; dDepartment of Botany, Lady Brabourne College, Kolkata, India;
e
Molecular and Applied Mycology and Plant Pathology Laboratory, Centre of Advanced Study, Department of Botany, University of Calcutta,
Kolkata, India; fCenter for Research in Nanoscience & Nanotechnology, University of Calcutta, Kolkata, India
ABSTRACT ARTICLE HISTORY

Copper and Copper oxide nanoparticles have garnered a lot of attention among the metal oxide Received 21 September 2021
nanoparticles, especially because of their many characteristics and applications in many Accepted 31 December 2021
disciplines, notably nanomedicine and biomedical sciences. We have covered all of the
KEYWORDS
conceivable green production techniques of copper/copper oxide nanoparticles in this review. Anticancer activity;
This manuscript also diagrammatically depicts the exact mechanism of all conceivable Antimicrobial activity;
biosynthetic routes. We also look at the antibacterial, antifungal, antiviral, and anticancer Copper/Copper oxide
properties of biosynthesized copper/copper oxide nanoparticles, as well as their effects on plant nanoparticles; Green
growth, nutrition, and defense mechanism. synthesis; Plant defense
Abbreviations: Cu/CuO NPs: Copper/Copper Oxide nanoparticles; DLS: Dynamic Light Scattering;
EDS: Energy Dispersive X-ray Spectroscopy; EDX: Energy dispersive X-Ray; FFT: Fast Fourier
transform; FTIR: Fourier transform infrared spectroscopy; HR-TEM: High-resolution transmission
electron microscopy, HeLa: Henrietta Lacks; IR: infrared; LSV: Linear sweep voltammetry; NTA:
Nanoparticle tracking analysis; PPE: Personal protective equipment; PSA: Particle size analyzer;
SEM: Scanning electron microscope; SPR: Surface plasmon resonance; TEM: Transmission
electron microscopy; VEGF: Vascular endothelial growth factor; XRD: X-ray diffraction.
1. Introduction
to synthesize nanomaterials and the synthesized nanoma-
In the sphere of research, nanotechnology is a relatively terials are simultaneously used to improve the quality of
new approach. This technology is now widely used (1) in life of the organisms (2). Biological synthesis uses the bio-
diversified fields. The smaller dimension of nanomater- logical principle of oxidation and reduction by microbial
ials, ranges from 1–100 nanometers (nm) (1), alters enzymes or plant phytochemicals (3). In recent times phys-
their physicochemical properties like shape, size, and ical and chemical methods are mainly used for the syn-
chemical composition. In the twenty-first century, a thesis of inorganic NPs (2). Both physical and chemical
more in-depth investigation of metallic NPs was methods have some disadvantages like low-productivity,
carried out by several researchers (2). non-eco-friendly, toxic, and capital intensive. For these
Bionanotechnology is a rapidly growing field of nano- reasons, biological synthesis is trying to replace the chemi-
technology in which bio-organisms are extensively used cal methods of producing NPs (4–23).
CONTACT Joy Sarkar [email protected] Department of Botany, Dinabandhu Andrews College, Garia, Kolkata, West Bengal 700084, India
© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.
188 N. CHAKRABORTY ET AL.
Numerous reports and studies have demonstrated material for stabilization (37). Additionally, this synthesis
that this green synthesis process has already manufac- route is straightforward, cost-effective, dependable, sus-
tured a large number of metal/metal oxide nanoparticles tainable, and relatively repeatable, and results in more
such as silver (Ag), gold (Au), selenium (Se), platinum stable compounds. Thus, researchers have expressed
(Pt), zinc oxide (ZnO) and iron oxide (Fe2O3), graphene an interest in developing a variety of nanomaterials via
oxide etc. (4–22). Additionally, those studies reported this biosynthesis route, including metal/metal oxide
that several metal nanoparticles have a variety of bio- nanoparticles, hybrid materials, and bioinspired
logical and biochemical activities, but CuNPs have materials. As a result, green synthesis is widely regarded
recently gained attention. Copper plays a variety of as a necessary tool for mitigating the negative conse-
roles in humans, including serving as a cofactor for quences of conventional nanoparticle synthesis
numerous enzymes involved in neuropeptide pro- methods used in laboratories and industries (38). In
duction, cell signaling pathway regulation, antioxidant that context, traditional nanoparticle producing tech-
defence, and immune cell function (24, 25). Copper is niques like chemical and physical synthesis are found
needed by plants for a variety of metabolic and physico- to be costly, hazardous, and unfriendly to the environ-
chemical processes. It’s one of the most crucial trace ment (39). Not only that chemical synthesis of nano-pro-
elements for plant growth (26). It is present in both duction may sometime affect biological activities by
humans and plants in a very small amount and helps some factors like size distribution, morphology, surface
to regulate different metabolic and biological activities charge, surface chemistry, capping agents, etc. (40, 41).
as it acts as cofactors for different enzymes. It is respon- To avoid the harmful effect, researchers have discovered
sible for the normal functioning of different essential the precise green pathways, or naturally occurring
proteins/enzymes such as amino oxidase, cytochrome sources and their products, that may be utilized to syn-
c oxidase, and plastocyanin since it is a cofactor for mul- thesize nanoparticles, to address these issues. To circum-
tiple enzymes (27, 28). Copper oxide, on the other hand, vent these negative consequences, researchers have
has antimicrobial, antibacterial, antifungal, antimicro- identified the precise green pathways, or naturally
bial, antifungal, magnetic phase change, gas sensing, occurring sources and their products, that can be
biocidal, superconductive, catalytic, and optical proper- employed to manufacture nanoparticles.
ties (29). CuO has a bandgap of 1.7 eV and is a p-type Nanoparticle synthesis can be divided into two cat-
semiconductor. The application of biologically syn- egories (Figure 1): (i) Top down method and (ii)
thesized copper nanoparticles (CuNPs) was found to bottom up method. The physical route of nanoparticle
be a promising bioactive agent in this context. synthesis is emphasized in the top down method,
CuONPs are made by different physicochemical whereas the chemical and biological methods are
methods like sol–gel technique (30), sonochemical (31), emphasized in the bottom up method.
electrochemical method (32), microwave irradiations Physical synthesis route consists of pulsed laser abla-
(33), solid-state reaction method (34), alkoxide based tion, arc discharge, spray pyrolysis, ball milling, vapor
route (35) etc. Likewise, these nanoparticles are also gen- and gas phase, pulsed wire dischare, lithography etc.
erated by the algal, fungal, plant and other biosynthesis On the other hand, chemical synthesis route consists
routes. In this review, we have focused on almost all the of chemical reduction, sonochemical, micoemulsion,
biosynthetic routes of CuNPs/CuONPs. The detailed photochemical, electrochemical, pyrolysis, microwave,
mechanism of all the possible biosynthetic routes is dia- solvothermal, coprecipitation etc.
grammatically represented in this manuscript. On the Additionally, green nanoparticle synthesis can be
other hand, high catalytic and chemical reactivity, divided into the following categories (Figure 1):
large surface area, and ability to interact with microbe’s
cells are some of the attributes of CuNPs/CuONPs which (a) Phyto routes like utilization of plants and plant
enables their application in different fields like agricul- extracts
tural, biomedical, textile, and environmental sectors (b) Microbial routes like the utilization of microorgan-
(36). Our review also focuses on the potential appli- isms such as fungi, yeasts (eukaryotes), bacteria,
cations of biosynthesized CuNPs/CuONPs. and actinomycetes
(c) Bio-template routes like the utilization of mem-
branes, viruses, and diatoms as templates
2. Green synthesis of nanoparticles
Green synthesis can be defined as the derivation of Both extracellular and intracellular biological
materials from green or eco-friendly resources by the approaches have been employed to synthesize nanopar-
use of solvent, good reducing agent, and harmless ticles. Although the precise method for the creation of
GREEN CHEMISTRY LETTERS AND REVIEWS 189
in the sample extract serve as a capping agent and

also aid in the stabilization of the produced nanoparti-
cles (43).
2.2. Green synthesis of Cu/CuO-NPs by plant

extracts
The environmentally accepted ‘green chemistry’ idea
has been applied to the biosynthesis of nanoparticles
for the creation of clean and environmentally friendly
nanoparticles, which incorporates bacteria, fungi,
plants, actinomycetes, and other organisms, and is
referred to as ‘green synthesis’. Biosynthesis of nanopar-
ticles utilizing the organisms mentioned above exem-
plifies a green alternative for the creation of
nanoparticles with novel characteristics. Unicellular and
multicellular organisms are permitted to respond in
these syntheses.
Plants are renowned as nature’s chemical factories
since they are low-cost and low maintenance. Because
extremely minute quantities of these heavy metals are
hazardous even at very low concentrations, plants
have shown exceptional potential in heavy metal detox-
ification as well as accumulation, through which
Figure 1. Various routes of nanoparticles synthesis (Created
with BioRender.com). environmental contaminants may be overcome (44–
50). Nanoparticle synthesis using plant extract has
benefits over other biological synthesis methods, such
nanoparticles utilizing biological agents has yet to be as microorganisms, because the rate of metal nanoparti-
discovered, it is suggested that distinct biomolecules cle synthesis with the help of plant extract is more per-
are involved for nanoparticle synthesis. Furthermore, sistent (51), significantly faster (52, 53), and extremely
the methods for intracellular and extracellular nanopar- mono-dispersive (54) in respect to other biological
ticle formation differ, and it appears that the cell wall of methods (55). The main challenges for using microor-
microorganisms plays a key role in intracellular nanopar- ganisms include the toxicity of certain bacteria, the iso-
ticle synthesis whereas extracellular enzymes play a key lation procedure of microorganisms, and the tedious
role in extracellular nanoparticle synthesis. Extracellular incubation procedure which make them unsuitable for
nanoparticle synthesis has surpassed intracellular nano- many researchers. Plant extracts are therefore a remark-
particle synthesis in popularity due to its faster pro- able source of synthesis of metal and metal oxide nano-
duction rate and simpler synthesis procedure (42). particles (55–57). Additionally, the reaction kinetics of
plant-assisted nanoparticle synthesis is much faster
than other biosynthetic methods that are comparable
2.1. Probable mechanism of biosynthesis of Cu/
to chemical nanoparticle production. Plant components
CuO-NPs
such as fruit, leaf, stem, and root have been frequently
Cu ions come in a variety of oxidation values, including employed for the green route of nanoparticle pro-
Cu(I), Cu(II), and a few Cu(III) ions. In terms of plant duction due to the high-quality phytochemicals they
extract, fungal extract, algal extract, bacterial extract, generate.
precursor concentration, pH, and temperature, the syn- Here, for the mentioned reason, Copper oxide nano-
thesis technique reported for CuO, Cu2O, and Cu4O3 is particles have been widely synthesized using various
the same to date. These variables, however, have the plant extracts (58–60). In this plant-based manufacturing
greatest impact on the type of Cu particles generated process, the metal salt is mixed with the plant extracts,
during green synthesis (43). During the green synthesis, and the reaction takes 1–3 h to complete at room temp-
the biomolecules present in the sample extract reduce erature (Figure 2). Plant extracts include a variety of bio-
the Cu2+ ion to Cu0 state and simultaneously oxidize it active metabolites, including flavonoids, phenols,
to form CuO nanoparticles. Certain biomolecules found proteins, terpenoids, and tannins, which serve as
Figure 2. Graphical representation of biomediated synthesis of copper oxide nanoparticles using plant extract (Created with
BioRender.com).
reducing and stabilizing agents, transforming metallic well established that when bacteria are introduced to
ions into nanoparticles (38, 44, 61). The plant extract pro- an environment containing high levels of hazardous
duces electrons, which cause copper salts to get metals, they can survive by converting harmful metal
reduced. Copper oxide nanoparticles are formed when ions to non-toxic metal oxides (128–131). Bacteria have
phytochemicals react with copper ions, resulting in been shown to generate a variety of essential thiol-con-
reduction. Table 1 summarizes the major contributions taining chemicals in response to oxidative stress. These
of biomediated synthesis of copper oxide nanoparticles molecules function as a capping agent in the bacterially
using various plants. driven production of nanoparticles, preventing metal
oxide nanoparticles from oxidizing (128, 132, 133). The
mechanism behind the nanoscale change isn’t fully
understood to date. Nanoparticle production also
2.3. Green synthesis of Cu/CuO-NPs by using
requires moderate experimental parameters such as
bacteria
pH, temperature, simple downstream processing, and a
Bacteria have been used to make a variety of nanoparti- short creation period (134). Some of the contributions
cles in recent years, including copper oxide nanoparti- of biomediated synthesis of copper oxide nanoparticles
cles (127). Different materials with fascinating shapes using different bacteria are shown in Table 2.
and nanoscale dimensions have been produced using
bacteria via an intracellular or extracellular route
(Figure 3). Bacteria have a great potential for nanoparti-
cles production. They offer benefits such as a short gen-
fungi
eration period, ease of culture, benign experimental
conditions, excellent stability, extracellular nanoparticle Various fungal species have been utilized to synthesize
synthesis, and ease of genetic modification (37). It is copper oxide and other metal nanoparticles in recent
known that when microorganisms are maintained in a years (127). Fungi, as compared to other microbes,
hazardous metal environment, they develop a method have a lot of potential for nanoparticle production. In
to live by converting poisonous metal ions into non- comparison to bacteria, fungi tolerate agitation, flow
toxic forms such as metal sulfide/oxides. It has been pressure, and other conditions in the bioreactor or any
Table 1. Biosynthesis of Cu/CuO nanoparticles by different plants.
Sl. Size of NPs Shape / structure /
No. Plants/ plant extract Precursor Part used (in nm) morphology Characterization techniques used Reference
1. Carica papaya L. Copper Chloride (CuCl2) Leaves 20 (in avg.) Nearly spherical and Crystalline UV-Vis, XRD, FTIR, SEM, and TEM (62)
2. Citrus medica L. Copper (II) Sulfate Pentahydrate (Cu Fruit 20 (in avg.) Crystalline UV-Vis, NTA and XRD (63)
SO4.5H2O)
3. Euphorbia esula L. Copper Sulfate (CuSO4) Leaves 20–110 Spherical UV-Visible spectroscopy, XRD, FTIR (64)
4. Pistacia sp. Cupric Chloride Dihydrate Leaves 9 (in avg.) Crystallized nanowires with 10 XRD, FTIR and SEM (65)
nm diameter
5. Nerium oleander L. Copper Sulfate (CuSO4) Leaves 21 Spherical UV-Vis and FTIR (66, 67)
6. Syzygium aromaticum (L.) Merr. & Copper Sulfate (CuSO4) Flower 14–50 Spherical and granular nature UV-visible spectroscopy, XRD, TEM and (66, 68)
L.M.Perry SEM
7. Ocimum sanctum L. Copper (II) Sulfate Pentahydrate Leaves 77 (in avg.) Spherical XRD and FTIR (66)
(CuSO4.5H2O)
8. Ocimum tenuiflorum L. Copper (II) Sulfate Pentahydrate Leaves 72 Hexagonal wurtzite crystal UV-visible spectroscopy, XRD, FTIR, SEM (66)
(CuSO4.5H2O) structure and EDAX
9. Citrus limon (L.) Osbeck Copper Chloride (CuCl2) Fruits 60–100 Nearly spherical UV-Vis, FTIR, XRD, SEM and TEM (69, 70)
10. Malva sylvestris L. Copper Chloride Dihydrate Leaves 14 (in avg.) Spherical XRD, FTIR and SEM (66)
(CuCl2.2H2O)
11. Albizia lebbeck (L.) Benth. Copper Sulfate (CuSO4) Leaves 100 (in avg.) Roughly spherical UV-Vis, SEM, TEM, EDS and XRD (66)
12. Datura metel L. Copper (II) Sulfate Pentahydrate Leaves 15–20 Spherical UV-Vis, PSA, TEM, EDX and FTIR (71)
(CuSO4.5H2O)
13. Magnolia kobus DC. Copper (II) Sulfate Pentahydrate Leaves 37–110 Spherical UV-Vis, ICP, EDS, XPS, and HR-TEM (72)
(CuSO4.5H2O)
14. Ginkgo biloba L. Cupric Chloride Dihydrate Leaves 15–20 Spherical UV-Vis, TEM, EDS, and FTIR (73)
(CuCl2.2H2O)
15. Artabotrys odoratissimus R.Br. Copper (II) Sulfate Pentahydrate Leaves 135 (in avg.) Spherical PSA (74)
(CuSO4.5H2O)
16. Capparis zeylanica L. Copper Sulfate (CuSO4) Leaves 50–100 Cubical structure UV-Vis, FTIR, XRD, SEM, EDX and TEM (75)
17. Aloe vera (L.) Burm.f. Copper (II) Acetate Dihydrate (Cu Flowers 40 (in avg.) Spherical UV-Vis, FE-SEM, and FTIR (76)
(CH3COOH) .2H2O)
18. Cinnamum sp. Copper Sulfate (CuSO4) Leaves, flowers 18.17-91.28 Spherical UV-Vis and DLS (77)
and roots
19. Inula helenium Copper Sulfate (CuSO4) Leaves, flowers 32.41 Spherical UV-Vis and DLS (77)

and roots
20. Matricaria chamomilla L. (Vernacular Copper Sulfate (CuSO4) Leaves, flowers 58.77 Spherical UV-Vis and DLS (77)
name: Chamomile) and roots
21. Urtica sp. Copper Sulfate (CuSO4) Leaves, flowers 6.5 Spherical UV-Vis and DLS (77)
and roots
22. Glycyrrhiza glabra L. Copper Sulfate (CuSO4) Leaves, flowers 28.21 Spherical UV-Vis and DLS (77)
and roots
23. Schizandra chinensis (Turcz.) Baill. Copper Sulfate (CuSO4) Leaves, flowers 32 Spherical UV-Vis and DLS (77)
and roots
24. Eucalyptus sp. Copper Sulfate (CuSO4) Leaves 38.62 (in avg.) Spherical UV-Vis, FTIR, XRD and SEM (78)
25. Zingiber officinale Roscoe Copper Sulfate (CuSO4) Rhizome 25–40 Spherical UV-Vis and XRD (79)
26. Eupatorium glandulosum Michx. Cupric Nitrate Leaves 55.91 (in avg.) Spherical UV-Vis, FTIR, PSA, TEM and AFM. (66)
27. Psidium guajava L. Cupric Chloride Dihydrate (Cu Leaves 13.13 ± 0.19 Spherical UV-Vis (66)
Cl2.2H2O)
28. Glycine max (L.) Merr. Copper Sulfate (CuSO4) Seeds 20 (in avg.) Spherical UV-Vis, TEM and DLS (66)
29. Bacopa monnieri (L.) Wettst. Copper (II) Sulfate Pentahydrate Leaves 20–50 Spherical UV-Vis, FTIR and HR-TEM (80)
(CuSO4.5H2O)
191
(Continued)
Table 1. Continued.
192
30. Ocimum basilicum L. Copper (II) Sulfate Pentahydrate Leaves 40–60 Spherical UV-Vis, FTIR and HR-TEM (80)
N. CHAKRABORTY ET AL.
(CuSO4.5H2O)
31. Asparagus adscendens Roxb. Copper (II) Sulfate Pentahydrate Leaves 10–15 Spherical UV-Vis, FTIR and HR-TEM (80)
(CuSO4.5H2O)
32. Withania somnifera (L.) Dunal Copper (II) Sulfate Pentahydrate Leaves 50–60 Spherical UV-Vis, FTIR and HR-TEM (80)
(CuSO4.5H2O)
33. Phyllanthus emblica L. Copper Sulfate (CuSO4) Fruits 15–30 Spherical UV-Vis, FTIR, XRD, SEM, EDAX
34. Sterculia urens Roxb. (Vernacular name: Cupric Chloride Dihydrate Gum 4.8 ± l.6 Spherical XRD, FTIR, TEM, SEM and XPS (66)
Gum karaya)
35. Gymnema sylvestre (Retz.) R.Br. ex Sm. Copper Sulfate (CuSO4) Leaves 65–302 Spherical UV-Vis, FTIR and SEM (81)
36. Ixora coccinea L. Copper Sulfate (CuSO4) Leaves 80–110 Spherical XRD, FTIR, SEM, TEM, (82)
37. Moringa oleifera Lam. Copper Sulfate (CuSO4) Leaves 6–61 Spherical XRD, FTIR, SEM, TEM, (83)
38. Tridax procumbens (L.) L. Copper Sulfate (CuSO4) Leaves 16 (in avg.) Spherical UV-Vis, XRD, FTIR, SEM, TEM (84)
39. Camellia sinensis (L.) Kuntze Copper Sulfate (CuSO4) Leaves 50–100 Spherical UV-Vis, FTIR, XRD, SEM, (85)
40. Juglans regia L. Copper Sulfate (CuSO4) Leaves 80 (in avg.) Spherical UV-Vis, FTIR, XRD, SEM, (86)
41. Acalypha indica L. Copper Sulfate (CuSO4) Leaves 26–30 Spherical UV-Vis, FTIR, XRD, SEM, EDX, and TEM (87)
42. Punica granatum L. Copper Aacetate Monohydrate [Cu Peel extract 40 (in avg.) Spherical UV-Vis, XRD, FTIR, SEM (88)
(CH3COO)2.H2O]
43. Musa acuminata Colla Copper Nitrate Trihydrate Solution (Cu Peel extract 60 (in avg.) Spherical XRD, EDX, FE-SEM, FTIR (89)
(NO3)2.3H2O)
44. Cordia sebestena L. Copper (II) Nitrate Trihydrate Solution Flowers 20–40 Spherical FESEM-EDX, XRD, FTIR, SEM, TEM (90)
(Cu(NO3)2.3H2O)
45. Hibiscus rosa-sinensis L. Copper Acetate Solution (Cu Flowers 26.54 Spherical UV-Vis, XRD, FTIR, and SEM (91)
(CH3COO)2.H2O)
46. Caesalpinia pulcherrima (L.) Sw. Copper (II) Nitrate (Cu(NO3)2.XH2O) Flowers 20 (in avg.) Spherical UV-Vis, FTIR, XRD, EDAX and SEM. (92)
47. Rheum palmatum L. Copper Chloride (CuCl2) Roots 30 (in avg.) Spherical UV-Vis, EDX, XRD, FTIR, SEM, TEM (93)
48. Desmodium gangeticum (L.) DC. Copper Sulfate (CuSO4) Roots 12 (in avg.) Spherical UV-Vis, TGA, XRD, FTIR, TEM, and SEM (94)
49. Phaseolus vulgaris L. Copper Sulfate (CuSO4) Whole plant 26.6 (in avg.) Spherical XRD, Raman, FTIR, TEM, XPS, DLS, SEM, (95)
SAED, and EDX
50. Coffea arabica L. Copper Sulfate (CuSO4) Whole plant 262 (in avg.) Crystalline UV-Vis, FTIR, XRD, SEM (96)
51. Quercus sp. Copper Sulfate (CuSO4) Whole plant 20–30 Quasi-spherical FE-SEM, XRD, FTIR (97)
52. Ziziphus mauritiana Lam. Copper Sulfate (CuSO4) Whole plant 20–45 Spherical XRD, SEM, EDX, TEM (81)
53. Ferulago angulata (Schltdl.) Boiss. Copper (II) Acetate Monohydrate (Cu Whole plant 44 (in avg.) Spherical XRD, SEM, EDX, TEM (98)
(CH3COO)2.H2O)
54. Gloriosa superba L. Copper Sulfate (CuSO4) Whole plant 5–10 Spherical XRD, SEM, EDX, TEM (99)
55. Syzygium alternifolium (Wight) Walp. Copper Sulfate (CuSO4) Bark 17.2 (in avg.) Spherical UV-Vis, XRD, FTIR, DLS, Zeta, TEM (100)
56. Zea mays L. Copper Sulfate (CuSO4) Dry husk 36–73 Spherical XRD, HR-TEM, EDX, FTIR (101)
57. Caesalpinia bonducella (L.) Fleming Copper Sulfate (CuSO4) Seeds 13.07 (in avg.) Spherical UV-Vis, XRD, FTIR, and SEM (102)
58. Vitis vinifera L. (Vernacular name: Copper Sulfate (CuSO4) Fruits 25–50 Uniform spherical UV-Vis, FTIR, XRD and SEM (103)
Erzincan Cumin)
59. Cedrus deodara (Roxb. ex D.Don) G.Don Copper Sulfate (CuSO4) Aqueous extract 16 Spherical FTIR, UV-Vis, XRD, TEM (104)
60. Abutilon indicum (L.) Sweet Copper (II) Nitrate Trihydrate (Cu Leaves 16.78 Spherical XRD, EDX, UV-Vis, SEM (105)
(NO3)2.3H2O)
61. Aloe barbadensis Mill. Copper Sulfate (CuSO4) Leaves 20 (in avg.) Spherical UV-Vis, SEM, TEM, XRD, FTIR (106)
62. Ficus religiosa L. Copper Sulfate (CuSO4.5H2O) Leaves 577 (in avg.) Spherical FE-SEM, UV-Vis, XRD, FTIR, DLS (107)
63. Phoenix dactylifera L. Copper Sulfate (CuSO4) Leaves 20–28 Spherical UV-Vis, FTIR, XRD, SEM, and EDAX (108)
64. Centella asiatica (L.) Urb. Copper Chloride (CuCl2.2H2O) Leaves 5 (in avg.) Spherical UV-Vis, IR, EDX (109)
65. Azadirachta indica A.Juss. Copper Chloride (CuCl2·2H2O) Leaves 38 (in avg.) Spherical UV-Vis, FTIR, XRD, SEM, EDX, DLS and (110)
TEM
(Continued)
Table 1. Continued.
66. Drypetes sepiaria (Wight & Arn.) Pax & Cupper Nitrate (Cu(NO3)2.3H2O) Leaves 298 (in avg.) Spherical UV-Vis, IR, XRD, and TEM, FTIR (111)
K.Hoffm.
67. Enicostemma littorale Blume Copper Sulfate (CuSO4) Leaves 30 (in avg.) Spherical UV-Vis, IR, XRD, and TEM, FTIR (112)
68. Cordia myxa L. Copper Sulfate (CuSO4.5H2O) Leaves 20–106 Spherical XRD, TEM, FTIR (113)
69. Arachis hypogaea L. Copper (II) Aetate Monohydrate Leaves 30–50 Spherical UV-Vis, XRD, FTIR, and SEM (114)
70. Leucaena leucocephala (Lam.) de Wit Copper Acetate Monohydrate (Cu Leaves 10–15 Spherical UV-Vis, XRD, FTIR, and SEM, BET (115)
(CH3COO)2.H2O)
71. Piper betle L. Copper Sulfate (CuSO4) Leaves 50–100 Spherical XRD, SEM, EDX, TEM (116)
72. Tabernaemontana divaricata (L.) R.Br. ex Copper Sulfate (CuSO4) Leaves 48 (in avg.) Spherical UV-Vis, FTIR, XRD, EDAX, TEM and SEM (117)
Roem. & Schult.
73. Ailanthus altissima (Mill.) Swingle Copper Sulfate (CuSO4) Leaves 20 (in avg.) Spherical UV-Vis, SEM, TEM, FTIR (118)
74. Saraca indica L. Copper Chloride (Cu Cl2.2H2O) Leaves 40–70 Spherical UV-Vis, XRD, EDX, FTIR, XPS, SEM, HR- (119)
TEM, TEM and SAED
75. Spinacia oleracea L. Copper Sulfate (CuSO4.5H2O) Leaves 1–12 Spherical XRD, TEM (120)
76. Eclipta prostrata (L.) L. Copper acetate Leaves 23–57 Spherical UV-Vis, FTIR, XRD, SEM, HR-TEM, EDS (121)
77. Cassia auriculata L. Copper Sulfate (CuSO4) Leaves 23 (in avg.) Spherical FTIR, UV-Vis, XRD, TEM, DLS (122)
78. Solanum lycopersicum L. Copper Sulfate (CuSO4.5H2O) Leaves 20–40 Spherical UV-Vis, FTIR, FE-SEM, HR-TEM, XRD, DLS (123)
79. Populus ciliata Wall. ex Royle Copper Nitrate Hexahydrate (Cu Leaves 50–60 Spherical FTIR, UV-Vis, EDX, SEM, XRD, TEM (124)
(NO3)3.6H2O)
80. Bauhinia tomentosa L. Copper Sulfate (CuSO4) Leaves 22–40 Spherical UV-Vis, XRD, TEM, EDX, FTIR (81)
81. Alternanthera sessilis (L.) R.Br. ex DC. Copper Sulfate (CuSO4) Leaves 22.6-25.2 Spherical SEM- EDAX, FTIR, XPS (81)

82. Citrofortunella microcarpa (Bunge) Copper Sulfate (CuSO4) Leaves 54–68 Spherical UV-Vis, FTIR, XRD, SEM and EDS (125)
Wijnands
83. Olea europaea L. Copper Sulfate (CuSO4) Leaves 20–50 Spherical UV-Vis, FTIR, XRD, TEM, and SEM (125)
84. Adiantum lunulatum Burm. f. Copper Sulfate (CuSO4) Whole Plant 1–20 Quasi-spherical UV-Vis, DLS, Zeta Potential, FTIR, XRD, (54)
TEM and EDX
85. Sida acuta Burm.f. Copper Sulfate (CuSO4) Leaves 50 Crystalline SEM,TEM, FTIR,Single beam (126)
spectroscopy,UV-vis
193
Figure 3. Graphical representation of biomediated synthesis of copper oxide nanoparticles using bacteria (Created with
BioRender.com).
other growth chamber. For the biogenic production of room temperature, which was then screened for
nanoparticles, cell-free extracts of microorganisms, func- copper extraction from integrated circuits and produced
tion as reducing, catalytic, or capping agents (127). Tri- in nano form. Dead biomass of Hypocrea lixii recovered
choderma species produce a wide range of bioactive from the metal mine was used to manufacture spherical
metabolites, including pyrones, polyketides, terpenes, CuNPs with an average size of 24.5 nm, and an infrared
diketopiperazine, glycolipids, and a large number of spectroscopy investigation was conducted, they discov-
reductive enzymes that aid in the production of not ered that amide groups in proteins were responsible for
just CuO nanoparticles but also Ag and ZnO nanoparti- the CuNPs stability and capping agents (158). Some of
cles (152–154). Fungi use both internal and external the contributions of biomediated synthesis of copper
routes to create various nanoparticles. The size of nano- oxide nanoparticles using different fungi are shown in
particles produced inside fungal species may be smaller Table 3 (Figure 4).
than those produced by the extracellular pathway, with
superior dispersity and dimensions. (155). Nanoparticle
fabrication via the extracellular route provides several
advantages. The nanoparticles created might be
algae
devoid of cell components. The extracellular route of
fungus has mostly been used to synthesize nanoparti- Algal members have gained importance in the synthesis
cles because fungi secrete several types of metabolites of CuONPs when these nanoparticles with sizes ranging
that function as reducing and stabilizing agents for from 5 to 45 and 6 to 7.8 nm have been effectively pro-
nanoparticle formation (154). Metal oxide nanoparticles, duced by utilizing a boiling aqueous extract from the
particularly copper oxide nanoparticles, have been syn- brown algae Bifurcaria bifurcata (170) and Cystoseira tri-
thesized using several fungus strains. nodis (171) respectively. Ramaswamy et al. (172) also
So, in this endeavor, several different species of employed an aqueous extract from brown seaweed (Sar-
fungus were studied, and it was discovered that fungi gassum polycystum) to make CuONPs. An autoclaved
are excellent candidates since they release huge aqueous extract from the green microalgae Botryococcus
amounts of enzymes and are easier to work within the braunii generated CuONPs with sizes ranging from 2–
laboratory. Penicillium aurantiogriseum, Penicillium citri- 10 nm (173). Alternatively, Bhattacharya et al. (174)
num, and Penicillium waksmanii all produce CuNPs extra- used a slightly different technique, heating the extract
cellularly (156). Majumder (157) described the at 50°C rather than boiling it, to get an aqueous
production of CuNPs from Fusarium oxysporum at extract from the microalgae, Anabaena cylindrica and
Table 2. Biosynthesis of Cu/CuO nanoparticles by bacteria.

Size of Shape /
Sl Gram Mode of NPs (in Structure / Characterization
No. Bacterial Culture Nature Precursor Synthesis nm) Morphology techniques used Reference
1. Escherichia coli (Migula) Gram- Copper Sulfate Extracellular 100– Quasi-spherical TEM, SEM, XRD, FTIR (135–137)
Castellani and Chalmers negative (CuSO4) 150
2. Mycobacterium Gram- Copper Sulfate Extracellular 15–20 Quasi-spherical SPR, FFT, UV-Vis, XPS, (138)
psychrotolerans Trujillo negative (CuSO4) cubic HR-TEM, LSV
and Morganella morganii
RP42 Winslow
3. Pseudomonas fluorescens Gram- Copper Sulfate Extracellular 20–80 Spherical and TEM, SEM, EDS, UV-Vis, (139)
Migula negative (CuSO4) hexagonal SPR
4. Pseudomonas stutzeri Gram- Copper Sulfate Extracellular 8–15 FCC, spherical TEM, SEM, EDS, UV-Vis, (140)
(Lehmann and Neumann) negative (CuSO4) SPR
Sijderius
5. Pseudomonas stutzeri Gram- Copper Sulfate Extracellular 50–150 FCC, cubic TEM, SEM, EDS, UV-Vis, (141)
(Lehmann and Neumann) negative (CuSO4) SPR
Sijderius
6. Serratia sp. Gram- Copper Sulfate Extracellular 10–30 Polydisperse TEM, SEM, EDS, UV-Vis, (142)
negative (CuSO4) SPR, EDX analysis
7. Streptomyces sp. Gram- Copper Sulfate Extracellular 100– Spherical TEM, SEM, EDS, UV-Vis, (143)
negative (CuSO4) 150 SPR, EDX analysis
8. Streptomyces cyaneus Gram- Copper (II) sulfate Extracellular 29.8 Spherical UV-Vis, DLS, FTIR, TEM, (136, 144)
(Krassilnikov) Waksman positive pentahydrate XRD
(CuSO4 ·5H2O)
9. Shewanella loihica PV-4 Gram- Copper (II) chloride Extracellular 6–20 FCC, cubic TEM, EDX, XRD, XPS (145)
negative dihydrate (CuCl2
·2H2O)
10. Salmonella typhimurium Gram- Copper (II) sulfate Extracellular 49 Spherical UV-Vis, DLS, SEM (146)
negative pentahydrate
(CuSO4 ·5H2O)
11. Bacillus cereus Gram- Copper (II) sulfate Extracellular 11–33 Spherical UV-Vis, DLS, Zeta, FTIR, (147)
positive pentahydrate EDX, SEM, TEM, AFM,
(CuSO4 ·5H2O) XRD
12. Streptomyces sp. MHM38 Gram- Copper (II) sulfate Extracellular 1.72– Spherical UV-Vis, EDX, TEM, XRD (148)
positive pentahydrate 13.49
(CuSO4 ·5H2O)
13. Lactobacillus casei subsp. Gram- Copper (II) sulfate Extracellular 30–75 Spherical FTIR, XRD, FESEM, TEM (149)
casei positive pentahydrate
(CuSO4 ·5H2O)
14. Morganella sp. Gram- Copper (II) sulfate Extracellular 15–20 Spherical UV-Vis, TEM, HR-TEM, (150)
negative pentahydrate XPS
(CuSO4 ·5H2O)
15. Actinomycetes Gram- Copper (II) sulfate Extracellular 61.7 Spherical UV-Vis, DLS, Zeta, FTIR, (151)
positive pentahydrate EDX, SEM, TEM, XRD
mycelial (CuSO4 ·5H2O)
effectively able to generate CuONPs from that extract using scanning electron microscopy, transmission elec-
with a particle size of 3.6 nm. tron microscopy, dynamic light scattering, particle analy-
Thus, the numerous algal members responsible for zers, and field emission scanning electron microscopy,
the reduction and stabilization process using copper as while UV–visible spectroscopy, X-ray diffraction, Fourier
a promotor, as well as their diverse organic components, transform infrared spectroscopy, surface plasmon reson-
have yet to be identified in detail (Figure 5). As a result, ance, and energy-dispersive X-ray spectroscopy were
it’s vital to focus research on the usage of biomolecules used to analyze the elemental chemical compositions
in green synthesis on CuONPs to expand their biological of Cu/CuONPs (Figure 6) (178).
applications. These are listed in Table 4.
4. Applications of copper nanoparticles

3. Characterization of CU/CUONPs (CUNPs)
To establish the demand for the generation of nanopar- CuNPs have diverse scientific applications. They are very
ticles one researcher should go through a series of effective against different pathogenic microbes. A high
characterization. After the synthesis of NPs, the crystal concentration of CuNPs generates reactive oxygen
structure and chemical composition are the initial species in bacterial cells which eventually cause cell
stage in the characterization process (177). The size lysis. Moreover, CuNPs have exhibited anticancer and
and morphology of the Cu/CuONPs were investigated antifungal activities. Due to their antimicrobial activity,
Table 3. Biosynthesis of Cu/CuO nanoparticles by fungi.

Size of
Sl. Mode of NPs (in Shape / Structure / Characterization
No. Fungal Culture Precursor Synthesis nm) Morphology techniques used Reference
1. Fusarium oxysporum Metallic copper Extracellular 93–115 – TEM, SEM, UV- Vis (157)
Schltdl.
2. Hypocrea lixii Pat. Metallic copper Extracellular 24.5 (in Spherical TEM, SEM, UV-Vis (158)
avg.)
3. (a) Penicillium 89–250 Spherical TEM, SEM, DLS, UV-Vis, (156, 159)
aurantiogriseum Dierckx FTIR, AFM
(b) Penicillium citrinum Copper Sulfate (CuSO4) Extracellular 85–295 Spherical
Thom
(c) Penicillium waksmanii 79–179 Spherical
K.W. Zaleski
4. Stereum hirsutum (Willd.) Copper salts (CuSO4, Extracellular 4–5 monodispersed, TEM, FTIR, XRD, and Zeta (160)
Pers. CuCl2) spherical Potential
5. Rhodotorula mucilaginosa CuCl2 Extracellular 10.5 (in Spherical (158)
(A. Jörg.) F.C. Harrison avg.)
6. Aspergillus niger Tiegh. Copper Sulfate (CuSO4) Extracellular 5–100 Spherical TEM, SEM, DLS, UV-Vis, (161)
FTIR
7. Trichoderma harzianum Copper (II) Sulfate Extracellular 5–18 Dense agglomerate TEM, SEM, DLS, UV-Vis, (154)
Rifai pentahydrate and spherical XRD
(CuSO4·5H2O)
8. Aspergillus flavus Link Copper (II) Sulfate Extracellular 20 (in Spherical TEM, UV-Vis, XRD, FTIR, (162)
pentahydrate avg.) NTA
(CuSO4·5H2O)
9. Trichoderma asperellum Copper nitrate (Cu(NO3) Extracellular 110 (in Spherical FETEM, HR SEM (163)
Samuels, Lieckf. & 2. 3H2O) avg.)
Nirenberg
10. Aspergillus fumigatus Copper nitrate (Cu(NO3) Extracellular 8 (in Spherical UV-Vis, HRTEM, Zeta, (164)
Fresen. 2. 3H2O) avg.) FTIR, XRD, EDX
11. Neurospora crassa Shear & Copper(II) chloride Extracellular 10–20 Spherical TEM, SEM, FTIR, EDX, XRD (165)
B.O. Dodge (CuCl2)
12. Pestalotiopsis sp. Copper(II) chloride Extracellular 10–20 Spherical TEM, SEM, FTIR, EDX, XRD (165)
(CuCl2)
13. Myrothecium gramineum Copper(II) chloride Extracellular 10–20 Spherical TEM, SEM, FTIR, EDX, XRD (165)
Lib. (CuCl2)
14. Alternaria alternata (Fr.) Copper (II) Sulfate Extracellular 60–80 Spherical TEM, EDX, XRF, SDD (166)
Keissl. pentahydrate
(CuSO4·5H2O)
15. Botrytis cinerea Pers. Copper (II) Sulfate Extracellular 60–80 Spherical TEM, EDX, XRF, SDD (166)
pentahydrate
(CuSO4·5H2O)
16. Aspergillus oryzae (Ahlb.) Copper (II) Sulfate Extracellular 55 Spherical UV–Vis., TEM, DLS, XRD, (81)
Cohn pentahydrate EDX, SEM and FT-IR
(CuSO4·5H2O)
17. Aspergillus terreus (Ahlb.) Copper (II) Sulfate Extracellular 15.75 Spherical UV–Vis, FT-IR, XRD, TEM (167)
Cohn pentahydrate and SEM
(CuSO4·5H2O)
i. Penicillium chrysogenum Copper (II) Sulfate Extracellular 9.70 Spherical UV–Vis., XRD, FTIR, DLS, (168)
Thom pentahydrate EDX TEM, SEM
(CuSO4·5H2O)
ii. Pleurotus ostreatus Copper nitrate (Cu(NO3) Extracellular 10–190 Spherical UV–Vis, TEM, DLS, XRD, (169)
(Kalchbr.) Pilát 2. 3H2O) XRD and FT-IR
it is also used for food preservation and agricultural field 4.1. Antimicrobial activity
to draw protection against different pathogenic fungi
In the past few decades, metal and metal oxides nano-
and bacteria (179). Copper-based nano fertilizer and
particles are being used to treat different bacterial and
nano-insecticides promote growth and nutrients in
viral diseases. Nanoparticles based antibiotics and
crop plants. Copper-based bioremediation plays a
other drugs gained a special attraction since it is less
pivotal role in waste-water treatment and removal of
toxic, eco-friendly and exhibited potential disease
heavy metals from soil. Copper is a good conductor of
curing activity. Several reports demonstrated that Cu
electricity hence it is used as a super-conductor and
and CuONPs have tremendous antimicrobial activities
has a significant contribution to the modern electronic
against different pathogenic microbes. High concen-
field (180). In Figure 7, we have summarized the
trations of CuNPs are toxic to the different bacterial
different applications of CuNPs and also elaborate its
pathogens of both humans and plants (181). CuNP has
major application in the following context.
Figure 4. Graphical representation of biomediated synthesis of copper oxide nanoparticles using different fungi (Created with
BioRender.com).
some unique features like small size, high surface area, When bacterial cells come in contact with CuNPs, it
biocompatibility, high biological and chemical reactivity develops toxicity inside the bacterial cell which leads
which helps to kill bacterial cells efficiently. Bio fabri- to several malfunctions and ultimately kills the cells.
cated CuNPs possess antimicrobial activity against Due to the small particle size of the CuNPs it can easily
both gram-positive and gram-negative pathogenic bac- take entry inside the bacterial cell through the cell mem-
terial strains (182). Green synthesized Cu and CuONPs brane. The carboxylic and amines group present in the
exhibited potential antibacterial activity against Pseudo- bacterial cell membrane helps to attract the Cu ions
monas aeruginosa, Clostridium difficile, Staphylococcus efficiently. The toxicity of CuNPs greatly varies with the
aureus and Escherichia coli (36, 81). The fabrication of size and shape of the particles. (192). CuNPs accumulates
CuNPs using Gloriosa superba leaf extract showed inhi- reactive oxygen species which can disrupt the cell mem-
bition against gram-positive bacteria Staphylococcus brane and provide direct cellular toxicity (185). Copper
aureus and gram-negative bacteria Klebsiella aerogenes has a great redox potential which can act as an electron
(179). Green synthesized CuONP using Sida acuta leaf donor or acceptor by producing Cu ions. These ions are
extracts can be used commercially in the textile industry very toxic for bacterial cells and accumulate superoxides
as a potential antimicrobial agent. It was found that and hydroxyl radicals leading to oxidative stress. These
S. acuta coated CuONPs can inhibit both Gram positive ROS generation can interfere with the cellular process
and Gram negative bacteria when it was applied in the of bacteria like DNA replication, cell division and metab-
cotton fabrics (126). Besides antibacterial activity, the olism (36). CuNP mediated toxicity in bacterial cells pro-
antiviral activity of green synthesized CuNP has also motes degradation of mitochondria, ribosomes and
been confirmed. Green synthesized CuNPs using clove different proteins channels present in the bacterial cell
fruit extract can inhibit the Newcastle disease virus membrane. The exact mechanism of antimicrobial
(183). In the following table, we have listed the appli- activity is still under study. A probable antimicrobial
cation of CuNPs as an anti-microbial agent against mechanism of CuNPs is presented in the following
different microbes (Table 5). diagram (Figure 8).
Figure 5. (a) Graphical representation of the preparation of the algal protein extract for the synthesis of copper oxide nanoparticles
using different algae (Created with BioRender.com) (b) Graphical representation of biomediated synthesis of copper oxide nanopar-
ticles using different algae protein extract (Created with BioRender.com).
4.2. Antifungal activity and crop protection synthesized by using Chitosan exhibited antifungal
activity against tomato plant pathogen A. solani and
In the current medical sector, the fungal disease has
F. oxysporum (195). Chemically synthesized CuNPs are
become a severe health threat and many patients die
highly effective against Candida albicans (C.P. Robin)
every year especially those with a poor immune
Berkhout and Aspergillus flavus Link (196). In another
system. Though there are several antifungal drugs are
report CuNPs exhibited antifungal activity against
available in the market, but the development of drug
some destructive crop pathogens like Alternaria alter-
resistance is a severe threat. Several studies revealed
nata (Fr.) Keissl., Curvularia lunata (Wakker) Boedijn
that CuNPs can be used as an antifungal agent since
and Phoma destructiva Plowr. (197). Green synthesized
they can inhibit several pathogenic fungi in both
CuNPs using Syzygium alternifolium (Wight) Walp. are
humans and plants (193). However, more studies are
known to have antifungal activity against some plant
needed to ensure the exact antifungal mechanisms of
pathogens. CuNPs can be used in the formulation of
CuNPs and their future use as an antifungal agent.
nanofungicides. In a field study, it was found that Cu
Green synthesized CuNPs can control several pathogenic
based fungicides are more effective than other agro-
fungal strains like Fusarium oxysporum Schltdl., Alternaria
chemicals against tomato pathogen Phytophthora infes-
solani (Ellis & G. Martin) L.R. Jones, Aspergillus niger
tans (Mont.) de Bary (198). Since CuNP has potential
Tiegh., and Penicillium citrinum Thom (194). CuNPs
antimicrobial and antifungal properties it could be
Table 4. Biosynthesis of Cu/CuO nanoparticles by algae.

Sl Size of NPs Shape/ Structure / Characterization techniques
No. Algal extract Precursor (in nm) Morphology used Reference
i. Bifurcaria bifurcata R. Ross Copper Sulphate solution 5–45 Crystallite UV-Vis, TEM, FTIR, XRD (170)
(CuSO4.5H2O)
ii. Cystoseira trinodis (Forsskål) Copper Sulfate (CuSO4) 6–7.8 Crystallite TEM, SEM, XRD, FTIR, EDX, (171)
C. Agardh Raman, UV-Vis
iii. Botryococcus braunii Kützing Copper Acetate Cu 10–70 Cubical and Spherical UV-Vis, FTIR, SEM, X-Ray (173)
(CH3COO)2 Diffraction.
iv. Anabaena cylindrica Copper Sulphate solution 3.6 (in avg.) Crystallite XRD, XPS, EDX (174)
Lemmermann (CuSO4.5H2O)
v. Macrocystis pyrifera (L.) C.Ag. Copper Sulfate (CuSO4) 2–50 Spherical DLS, Zeta Potential, FTIR, TEM, (175)
EDS
vi. Sargassum polycystum Aqueous copper – Spherical TEM, SEM, XRD, FTIR, EDX, (176)
C. Agardh UV-Vis
Figure 6. Characterization of synthesized copper oxide nanoparticles (CuONPs) (a) TEM images of CuONPs (b) XRD analysis of CuONPs
(c) UV-Visible Spectroscopic analysis of CuONPs (d) FTIR analysis of CuONPs (e) EDX analysis of CuONPs.
used in food preservation and food packaging as well. studies are still under process to discover an alternative
High concentrations of copper provide direct toxicity nontoxic biological drug. In this regard, it was found that
to several food spoilage microbes and fungi (81). Some the emergence of nanotechnology helps to treat
antifungal applications of CuNPs are listed below different types of cancer efficiently. Biologically syn-
(Table 6). thesized Cu and CuO nanoparticles exhibited promising
results when it tested against some human cancer cell
lines (Table 7) (201–207).
4.3. Anticancer activity
Green synthesized CuNPs using dry black beans can
Currently, cancer is the most dangerous and common inhibit the growth of human cervical carcinoma and
disease which increases the mortality rate worldwide. also showed cytotoxicity against HeLa cell line by produ-
Till now no promising drugs are available in the cing ROS (204). The mechanism of CuNP mediated antic-
market to treat cancer. The most commonly used radio- ancer activity includes oxidative stresses, accumulation
therapy and chemotherapy have tremendous side of ROS, chromosomal aberration, genetic material frag-
effects and are also expensive processes. Several mentation, production of caspases, enhancement of
Figure 7. Applications of copper nanoparticles (Created with BioRender.com).
intrinsic and extrinsic apoptotic pathways (Figure 9) normal control cells depicted by a normal mitochondrial
(192). structure whereas the treated cancer cells are character-
In a study, it was found that green synthesized ized by a condensed clumped structure of mitochondria
CuONPs can inhibit the growth of A549 adenocarcinomic which leads to apoptosis of the cancer cells (83). When
human alveolar epithelial cells. A high concentration of the similar study was done with HepG-2 hepatic cancer
CuONPs promotes cellular toxicity and DNA damage in cell line it exhibited enhanced apoptosis by upregulating
the A549 lung cells (208). In addition to this, it was also the tumor suppressor gene and down regulating the
found that biologically synthesized chitosan/copper antiapoptotic gene bcl-2 (211).
oxide nanocomposite using a bioflavonoid rutin, also To explain this finding, it is important to note that
exhibited antiproliferative efficiency when it was tested various factors, including size distribution, shape, and
against A549 human lung cancer cell line (209). CuNP surface chemistry of biogenic nanoparticles, can
mediated anticancer activity has been observed in influence their cytotoxicity. Different cytotoxicity
CaCO-2 human colon cancer cells, Mcf-7 breast cancer responses may result from changes in various par-
cells, HepG-2 hepatic cancer cells and HeLa cells (210). ameters. Furthermore, biomolecules are responsible for
In a study, it was found that when HeLa cells are bioreduction of metal ions to their nano-forms in the
treated with copper oxide nanoparticles it showed oxda- biosynthetic method. These biomolecules are attached
tive stress mediated mitochondrial degradation. The to the surface of biosynthesized nanoparticles and act
Table 5. Applications of green synthesized CuNPs as antimicrobial agent.

Sl. Chemical
No. Biological source used Antimicrobial Application Reference
1. Eucalyptus globulus Labill. CuSO4 It accumulates ROS inside the bacterial cells which lead to cell destruction. (137)
2. Zea mays L. Cu It is effective against some pathogenic bacteria like Bacillus lichiniformis and Pseudomonas (184)
(CH3COO)2 aeruginosa.
3. Citrus medica L. CuSO4 It can inhibit several crop pathogens and also inhibit the growth of Propionibacterium acnes (63)
and Klebsiella pneumoniae.
4. Camellia sinensis (L.) Kuntze Cu(NO3)2 It can inhibit the growth of Vibrio cholerae and Klebsiella pneumonia. (185)
5. Bacillus cereus Frankland CuSO4 It can control the growth of pathogenic bacteria like P. aeruginosa, Staphylococcus aureus, (186)
and Frankland E. coli and Bacillus subtilis.
6. Nerium oleander L. CuSO4 Antibacterial activity against Salmonella typhi, E. coli, Bacillus subtilis and Staphylococcus (187)
aureus.
7. Sida acuta Burm.f. CuSO4 Exhibited tremendous antimicrobial activity against some human pathogens and also (126, 188)
effective against bacterial contamination in the textile industries.
8. Punica granatum L. CuSO4 Exhibited antimicrobial activity against Micrococcus luteus, Salmonella enteric and (189)
Enterobacter aerogenes.
9. Pseudomonas fluorescens CuSO4 Inhibit the growth of Bacillus and E. coli. (134)
Migula
10. Chitosan derived from CuSO4 Effective against Salmonella paratyphi. (190)
fungal cell wall
11. Ocimum sanctum L. CuSO4 Antimicrobial activity against human pathogenic microbes. (191)
4.4. Plant growth, nutrients and defense booster

Plants need to uptake different minerals and nutrients
elements for their proper growth and development.
Plants nutrients are divided into two classes macronutri-
ents and micronutrients. Macronutrients are generally
required in high concentrations and micronutrients are
required comparatively in low concentrations. Both
these macro and micronutrients are essential for main-
taining structural integrity and normal growth in
plants. Deficiency of which causes disease and death
of different plant parts (81, 216). In plants, copper is
required in very low concentrations as it is a micronutri-
ent. A high concentration of Cu promotes toxicity and
hampered growth in plants. Plant chloroplast contains
a maximum number of Cu as it helps in chloroplast
and other pigments synthesis. Cu deficiency leads to
several abnormal conditions like young leaf distortion,
necrosis, stem bending, affects vegetative growth and
reduces grain quality in crop plants (217). CuNP
mediated plant growth responses depend upon
several factors like concentration, size, plant species
and structure of the particles. In a study, it was found
that CuNPs can stimulate root and shoot growth in Pha-
seolus radiates and Triticum aestivum. The growth
response typically varies with varying concentrations of
CuNPs. When wheat plants were treated with 20, 25,
30 and 35 ppm concentrations of CuNPs it showed
better growth and yields. Concentrations above
Figure 8. Probable antimicrobial mechanism of Cu/CuONPs
(Created with BioRender.com). 1000 ppm reduced growth in wheat followed by a
decrease in yields (218). When CuNPs were applied on
Allium cepa with 20 µg/ml concentrations it enhanced
as a stabilizer, preventing the nanoparticles from aggre- growth and mitotic index. The mitotic index decreased
gating. These biomolecules adhering to the surface of with increasing concentrations of CuNPs (219). Appli-
the nanoparticles, have the potential to alter the cations of CuONPs increased shoot and root growth in
surface chemistry of various nanoparticles and interfere Zea mays (220). Application of CuONPs on transgenic
with their ability to respond to their biological environ- cotton plants enhanced expression of exogenous
ment. As a result, the biological sources used to make genes which code for Bt toxins in leaves (221). On the
nanoparticles may have an impact on their cytotoxicity contrary high concentration of CuNPs has a negative
response (212–215).
Table 6. Antifungal activity of different green synthesized CuNPs.

Sl. Chemical
No. Biological source used Antifungal Application Reference
1. Cissus quadrangularis L. Cu Inhibit the growth of pathogenic fungi Aspergillus flavus Link and A. niger Tiegh. (81)
(CH3COO)2
2. Brassica juncea (L.) Czern. CuSO4 Inhibit the growth of Alternaria alternata, Phoma destructiva and Curvularia lunata (81)
(Wakker) Boedijn.
3. Citrus medica L. CuSO4 Effective against pathogenic fungi Fusarium graminearum Schwabe, F. oxysporum (63)
Schltdl. and F. culmorum (Wm.G. Sm.) Sacc.
4. Oxalis corniculata L. CuSO4 Inhibit the growth of A. alternata, Pythium ultimum Trow and A. niger Tiegh. (199)
5. Syzygium alternifolium (Wight) Cu(NO3)2 Effective against Trichoderma harzianum Rifai (183)
Walp.
6. Penicillium chrysogenum Thom CuSO4 Antifungal activity against A. solani (Ellis & G. Martin) L.R. Jones, A. niger Tiegh. and (194)
F. oxysporum Schltdl.
7. Syzygium aromaticum (L.) Merr. & CuSO4 Exhibited antifungal activity against pathogenic fungi A. niger Tiegh. and A. flavus (200)
L.M.Perry Link.
Table 7. Different anticancer properties of CuNPs.

Sl. Tested cell
No. Biological source lines Anticancer activity Reference
1. Camellia sinensis (L.) Kuntze MCF-7 Promotes significant cytotoxicity against breast cancer cells. (174)
2. Bacillus cereus Frankland and A549 Inhibit the growth of the tested cancer cell line. (153)
Frankland CaCO-2
HCT 116
3. Ficus religiosa L. A459 Inhibit lung cancer cells by inducing ROS mediated apoptosis. (175)
4. Phaseolus vulgaris L. HeLa Inhibit the growth of human cervical cancer cells which includes ROS mediated (171)
apoptosis and mitochondrial malfunctions.
5. Syzygium alternifolium (Wight) MCF-7 Inhibit the growth of breast cancer cells by promoting direct cytotoxicity. (150)
Walp.
6. Cordia myxa L. AMJ-13 Inhibit breast cancer cell growth. (176)
MCF-7
7. Calotropis procera (Aiton) BHK 21 Promotes apoptotic destruction of the tested cancer cell lines. (177)
Dryand. HeLa
A549
8. Lactobacillus casei AGS Inhibit the growth of human gastric cancer cells and colorectal cancer cells by (178)
HT-29 promoting cytotoxicity and apoptosis.
impact on plant growth. CuNPs with different concen- 4.5. Wound healing
trations ranging from 200 to 1000 mg/l exhibited a
Several studies over the last few years have ensured that
negative impact on the growth of Raphanus sativus, Tri-
CuNPs can be used as wound healing agents providing
ticum aestivum, Lolium perenne and Phaseolus radiates
protection against infections. Wounds and cuts on the
(222). Application of CuONPs at a higher concentration
body surface are the most common pathway through
in Arabidopsis thaliana seedlings inhibited root and
which different infectious microorganisms take entry
shoot growth and also decreased chlorophyll contents
into the body. To prevent infection, it is very necessary
(223). Recently, we have shown that treatment of
to remove microbes from the wound site. As discussed
CuONPs significantly boost up the defense enzymes,
earlier, CuNPs has potential antimicrobial and antifungal
total phenol and other defense parameters along with
activity so it may be used in wound healing purposes
plant vigor in Lens culinaris through nitric oxide signal-
(225). At the site of the wound Cu stimulates the for-
ing pathway (224). Possible mechanism has been illus-
mation of new blood vessels which leads to angiogen-
trated in Figure 10.
esis. Besides this CuNPs also stimulates the expression
Figure 9. Probable anti-cancer mechanism of Cu/CuONPs (Created with BioRender.com).

Figure 10. Schematic representation of defense induction and plant growth promotion by Cu/CuONPs (Created with BioRender.com).
of vascular endothelial growth factor (VEGF) that facili- several pathogenic microbes (210). CuNPs can be used
tates the transport of different nutrients and collagen to treat polluted water in different industrial areas.
formation necessary for repairing the wound (Figure Several reports have described that CuNPs are used in
11). Application of Cu can halt severe phosphorus burn the disposal of industrial waste and effluent. Besides
by the process of tissue remodeling. At the site of the bioremediation activity, CuNPs has a potential biocataly-
wound Cu helps in maintaining the stability of fibrino- tic activity that can reduce and degrade xanthenes dyes,
gen and accumulates lysyl oxidase enzyme (210). congo red, rhodamin-B, and methylene blue (70). Due to
Green synthesized CuNPs using Falcaria vulgaris leaf fluorescence quenching properties, CuNPs could be
extracts exhibited potential cutaneous wound healing used as biosensing and biolabelling agent (228). Since
activity (226). In a study, a 500 mm cut was made on CuNPs has a very small size it can interact with
an anaesthetized animal. Three set up was made by different biomolecules efficiently and can be used as
applying biologically synthesized CuNP gel, nonbiologi- drug delivering system (179). Green synthesized CuNPs
cally synthesized CuNP and control. After some days it exhibited larvicidal activities against Aedes aegypti pro-
was found that biologically synthesized CuNPs were viding protection against dengue, zika and chikungunya
able to reduce 92% of the wound. Moreover, it promotes (229).
cell proliferation, cell migration and inhibits cyclooxy-
genase-2 enzyme at the wound site (186, 227).
5. Future perspectives
In this era of nanotechnology, remarkable advance-
4.6. Other applications
ments are going on day by day in the field of nanopar-
CuNPs are now used in the textile industry for making ticle synthesis and their sophisticated applications in
antimicrobial personal protective equipment (PPE). In the vast arena of science and technology. Beside
the fabric’s polymer, nano-coppers are incorporated for others CuONPs/CuNPs have tremendous widespread
making antimicrobial fabrics. CuNPs incorporated applications, especially in biological systems. However,
cotton fibers exhibited antimicrobial activities against some nanosystems needs further sophistication and
Figure 11. Probable wound healing mechanism of Cu/CuONPs (Created with BioRender.com).
they are still at the stage of infancy. More widespread the use of hazardous compounds in various technical
research is still required to set the parameters. New fields of application-based research. In this context, it
cost-effective tools must be developed. Searching for is evident to utilize CuONPs/CuNPs as a hazard free com-
new application of CuONPs/CuNPs has to be taken pound, at various important sectors of research like drug
into consideration. Application of CuONPs/CuNPs in delivery system, in solving various health issues, plant
the research field of nano-sensors in film packaging of defence booster, textile industry, etc. However, synthesis
food, detection of microorganisms and toxic substances part is very important as harmful substances are used in
to check food quality etc. will give future directions. both physical and chemical methods of CuONPs/CuNPs
However, challenges and safety aspects can be taken production. The biological technique, on the other
as a serious concern. The findings of this review hand, is eco-friendly, cost-effective, dependable, stable,
expand the prospects for the green synthesis of uses little energy, and is a straightforward procedure.
CuONPs/CuNPs and their use in a variety of biological In this study, a comprehensive idea has been given on
and biotechnological domains. the pros and cons of bio-based synthesis and their
characterization including their every possible appli-
cation. However, to advance the biomedical applications
6. Conclusion
of CuONPs, additional research should be conducted on
In this present situation of global pollution and environ- ways to reduce the toxicity of CuONPs while preserving
mental consciousness it is utmost important to reduce and enhancing their biological efficiency.
Acknowledgements [5] Sarkar, J.; Acharya, K. Alternaria Alternata Culture Filtrate

Mediated Bioreduction of Chloroplatinate to Platinum
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help received from the scholars whose articles are cited and 1186062.
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Disclosure statement Properties of the HPMC-γ-Fe2O3 Nanocomposites.
Bioprocess Biosyst. Eng. 2017, 40, 351–359. doi:10.1007/
No potential conflict of interest was reported by the author(s).
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Krishnendu Acharya http://orcid.org/0000-0003-1193-1823 Mangrove Fruit Polysaccharide for Bacterial Growth
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Green Synthesis of Coppercopper Oxide Nanoparticle

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GREEN CHEMISTRY LETTERS AND REVIEWS

2022, VOL. 15, NO. 1, 187–215

Green synthesis of copper/copper oxide nanoparticles and their applications:

ABSTRACT ARTICLE HISTORY

in the sample extract serve as a capping agent and

2.2. Green synthesis of Cu/CuO-NPs by plant

GREEN CHEMISTRY LETTERS AND REVIEWS

GREEN CHEMISTRY LETTERS AND REVIEWS

Table 2. Biosynthesis of Cu/CuO nanoparticles by bacteria.

4. Applications of copper nanoparticles

Table 3. Biosynthesis of Cu/CuO nanoparticles by fungi.

Table 4. Biosynthesis of Cu/CuO nanoparticles by algae.

Figure 7. Applications of copper nanoparticles (Created with BioRender.com).

Table 5. Applications of green synthesized CuNPs as antimicrobial agent.

4.4. Plant growth, nutrients and defense booster

Table 6. Antifungal activity of diﬀerent green synthesized CuNPs.

Table 7. Diﬀerent anticancer properties of CuNPs.

Figure 9. Probable anti-cancer mechanism of Cu/CuONPs (Created with BioRender.com).

Acknowledgements [5] Sarkar, J.; Acharya, K. Alternaria Alternata Culture Filtrate

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