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Fig. 8.3 Schematic illustration of actinomycetes-mediated green synthesis of nanoparticles
S. Paul et al.
physicochemical characterization of the nanoparticles was performed to determine
their structural and chemical properties. Figure8.3 provides a visual representation
of the bioprocess involved in nanoparticle synthesis mediated by actinomycetes.
8.4.3 Factors Affecting NP Biosynthesis
Research has indicated that the size of biosynthesized nanoparticles greatly affects
their biological activity. Smaller nanoparticles have high surface/volume ratios and
exhibit superior activity levels compared to larger NPs (Pandit etal. 2022). The
culture techniques employed in actinomycete-assisted NP synthesis play a signicant role in the synthesis of homogenous nanoparticles with improved enzymatic
activity. Therefore, optimizing culture parameters, including medium composition,
temperature, pH, mixing speed, and incubation time, can signicantly impact the
synthesis of nanoparticles with enhanced enzymatic activity (Kumari etal. 2020).
pH
pH is found to be an essential variable in the synthesis of biogenic NPs. Metal ion
reduction may be impacted by a sudden change in pH because it can alter the charge
of the metal ions in the reaction mixture. According to Patra and Baek 2014, the pH
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of the solution media can make a determent effect on the size and texture of synthesized nanoparticles (Patra and Baek 2014). An interesting nding in the biosynthesis of AgNPs was reported by Ibrahim etal. (2023), based on the observation that
alterations in the pH of the solution could modify the size of the NPs (Ibrahim etal.
2023). The samples produced at pH5 were unstable, whereas those synthesized at
pH9 were stable.
Temperature andPressure
Temperature is another crucial factor for NP synthesis. The green synthesis of
nanoparticles requires ambient temperature, and the temperature of the reaction
determines the type of nanoparticle created, which has a major impact on the rate of
synthesis, stability of NPs, and nal size of the NPs produced (Patra and Baek 2014).
Another signicant element that inuences the nal form of metallic NP size and
shape is pressure. Research has shown that ambient pressure accelerates the reduction of metal ions compared to normal conditions (Pandit etal. 2022).
Concentration
The concentration of the cell extract is also critical for efcient NP biosynthesis,
which relates the quantity of stabilizing and reducing agents (Kumari etal. 2020).
147
Reaction Time
Another crucial element for optimization during biosynthesis is the time necessitated to complete the bioreduction process among the metal precursor and extract.
The amount of time the reaction medium is incubated offers a signicant impact on
the quality and structure of the NPs produced. The chemical properties of NPs are
also altered according to the reaction time in a similar way. If nanoparticles are
incubated for an extended period, they may aggregate, shrink, and lose some efcacy (Gupta etal. 2019).
8.4.4 Characterization Techniques forMetal NPs
Several characterization approaches have been used to study the physiochemical
properties of nanoparticles. This covers a variety of analytical techniques, such as
FT-IR spectroscopy, NMR spectrometry, and UV-Vis spectrometry. Optical or Zeta
potential (Zp) measurements were performed using DLS, CD, and XRD.An array
of microscopic analyses by AFM, SEM, TEM, and STM are also used methodically
to analyze the various physical properties of NPs. Moreover, certain complementary
analytical procedures, such as MS, TERS spectrometry, and SERS spectrometry,
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S. Paul et al.
Fig. 8.4 Graphical representation of some of the physicochemical characterization and techniques
of nanoparticles. Abbreviations: NPs nanoparticles, ZP zeta potential, FTIR Fourier transform
infrared spectroscopy, FS uorescence correlation spectroscopy, NMR nuclear magnetic resonance
spectroscopy, UV-Vis ultraviolet-visible spectroscopy, CD circular dichroism, DLS dynamic light
scattering, XRD X-ray crystallography, AFM atomic force microscopy, SEM scanning electron
microscopy, STM scanning tunneling microscopy, TEM transmission electron microscopes, MS
mass spectrometry, SERS surface-enhanced Raman spectroscopy, TERS tip-enhanced Raman
spectroscopy
are also employed to study the details of the synthesized NPs (Fig.8.4) (Wadhwa
etal. 2022).
The process of investigation using absorption spectra in the range of 200–800nm
will help to conrm that nanoparticles were formed during synthesis. Techniques
such as SEM and TEM can be employed to analyze the size, shape, distribution, and
homogeneity of nanoparticles. The crystallinity and chemical composition of the
synthesized NPs were determined using XRD.FTIR spectroscopy was exercised to
identify the functional groups existing in the nanoparticles. DLS is an effective
method for estimating the particle size, even at low concentrations. The surface area
and stability of colloidal nanoparticles were examined using optical measurements
(Shnoudeh etal. 2019).
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8.5 Applications ofNPs inDifferent Biotechnological Sectors
Nanoparticles have demonstrated versatility for potential applications in various
biotechnological sectors, including agriculture, environment, food, and therapeutics. The utilization of metallic nanoparticles in biotechnology has increased because
of their inherent biocompatibility, potent antimicrobial properties, prociency in
drug delivery, profound bioactivity, enhanced bioavailability, and effective integration into biological systems. One unique characteristic of biosynthesized metalbased nanoparticles is their lack of requirement for external stabilizing or capping
agents, which has unlocked potential in previously unexplored applications in various domains (Zarina and Nanda 2014).
8.5.1 Applications ofNPs intheEnvironmental Sector
The environmental applications of nanotechnology primarily fall into three major
domains: NPs as environmentally sustainable products to combat pollution, their
use in bioremediation of toxic contaminants, and their function as biosensors for
environmental monitoring. Because of the detrimental effects of heavy metals on
both the environment and human health, NPs have garnered a considerable amount
of attention as potential biosorbent materials for the removal of heavy metals from
environmental sources. Metallic nanoparticles sourced from Actinomycetes have
proven to be powerful tools in the ght against biofouling, with silver nanoparticles produced by Streptomyces rochei HMM13 having the ability to reduce the
number of bacterial cells and prevent bacterial biolm formation (Sawada
etal. 2012).
8.5.2 Applications ofNPs inFood Sector
Nanotechnology has abundant applications in the food industry, including functional food development, food processing and packaging, food safety like in the
detection of food pathogen, and extending the shelf life of food products. These
applications can be broadly categorized into two groups: nanostructured food
ingredients and nanosensing. Nanostructured food components are used in food
processing, where they can be used as llers to increase the durability of packaging materials, enhancing mechanical strength and as food additives, anticaking
agents, and antimicrobial agents. Nanosensing can also be applied to enhance
food safety and quality assessments (Singh etal. 2017). A potential application of
green nanosynthesis in the food industry is the use of Streptomyces-mediated
coatings to create natural antibacterial food coatings. These materials have the
potential to prolong the shelf life of food products and ensure safety (Rajamanickam
etal. 2012).
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S. Paul et al.
8.5.3 Applications ofNPs inAgricultural Sector
The synthesis of green NPs has also emerged as a potential source of fertilizers,
insecticides, and agents for plant growth. In recent years, there has been a trend
toward using nanomaterials as substitutes for traditional pest management techniques in agriculture. Various nanomaterials and nanostructures are currently being
used as biosensors to assess soil quality and fertilizer distribution. According to
Karthik et al. 2014, extracellular AgNP synthesized by Streptomyces sp. LK3 is
particularly effective in controlling pests, particularly Haemaphysalis bispinosa and
Rhipicephalus microplus, in agriculture (Karthik etal. 2014).
8.5.4 Applications ofNPs inMedical Sector
Nanomedicine has garnered signicant attention in every branch of medicine, from
diagnostics to treatment. Its applications include antimicrobial agents, biomolecule
detection, biosensors, cancer treatment, diagnostic devices, drug delivery, regenerative medicine, and tissue engineering. The unique properties of nanomaterials, such
as enhanced exibility, durability, sensitivity, unparalleled performance, and distinct physicochemical properties, have been exploited in therapeutic applications
for early detection of diseases, regenerative medicine, targeted clinical therapy, and
tissue reconstruction. Nanomedicine operates in a manner similar to many other
biological processes and has fewer side effects (Rudramurthy and Swamy 2018;
Mauricio etal. 2018; Patel and Nanda 2015).
Actinomycetes can synthesize numerous metal nanoparticles, such as copper,
gold, silver, and zinc, which exhibit antimicrobial effects against a diverse array of
microorganisms, including multidrug-resistant strains (Table8.2). The versatility of
the antimicrobial activity against a wide range of microbes is noteworthy.
Researchers have reported that AgNPs synthesized by actinomycetes demonstrate
bactericidal activity against clinically signicant pathogens, both inside and outside
cells. For example, Streptomyces viridochromogenes exhibit excellent antimicrobial
activity against both gram-negative and gram-positive bacteria, as well as yeasts,
such as Candida albicans (Krishnaraj etal. 2016). Their small S/V ratio (size/volume) allows them to interact more effectively with microbial membranes, causing
the cells to rupture (Chauhan etal. 2013). The increasing emergence of antibiotic
resistance underscores the potential of actinobacteria-mediated NP as promising
alternatives. Another study explored the synergistic potential of AgNPs derived
from Streptomyces parvulus SSNP11. These nanoparticles can boost the effective-
ness of traditional antibiotics such as Ampicillin, Cefalexin, Penicillin, and
Streptomycin. Additionally, these NPs have the potential to serve as carriers for
drug delivery, which is crucial in the ght against microbial infections. The antiESBL properties of the AgNPs isolated from Streptomyces sp. VITSJK10 have also
been previously reported (Manikprabhu and Lingappa 2013). Biogenic ZnO oxide
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Table 8.2 Examples of actinomycetes-mediated NPs with therapeutic potential
Actinomycetes sp.NP(Size nm) Applications References
S. durhamensis Ag
(8–48)
N. sp. MBRC-1 Ag
S. rochei Ag
Streptomyces sp.. Ag
G. amicalis HS-11 Ag, au
Rhodococcus sp. Au
S. parvulus SSNP11 Ag
Rhodococcus sp. Au
S. xinghaiensis OF1 Ag
Streptomyces sp.
VITSTK7
Streptomyces sp. LK3 Ag
S. rochei HMM13 Ag
a
Synergistic effects with ciprooxacin, ampicillin, streptomycin, gentamicin, tetracycline, and
lincomycin; B Bacillus, E E. coli, K Klebsiella, P Pseudomonas, C Candida, H Haemaphysalis, R
Rhipicephalus, S Streptomycetes, N Nocardiopsis, G Gordonia, NP nanoparticle, Ag silver, Au gold
(45)
(22–85)
(21–45)
(−)
(10)
(2–12)
(9)
(5–20)
Ag
(20–60)
(−)
(−)
Antimicrobial (S. aureus, B.
subtilis, E. coli, P. aeruginosa,
K. pneumoniae,
and P. mirabilis)
Antimicrobial (B. subtilis, P
aeruginosa, and C. albicans)
Antibacterial & Antibiolm
Anticandidal Prakasham etal. (2014)
Free radical scavenging Kumari etal. (2020)
Antimicrobial and antioxidant Manivasagan etal. (2015)
Antibacterial Prakasham etal. (2014)
Antifungal Kumari etal. (2020)
Antimicrobial Manivasagan etal. (2015)
Antifungal Prakasham etal. (2014)
Acaricidal (H. bispinosa and R.
microplus)
Antibacterial and antibiolm Manivasagan etal. (2015)
Kumari etal. (2020)
Manivasagan etal. (2015)
a
Prakasham etal. (2014)
Karthik etal. (2014)
151
nanoparticles show potential for cancer treatment and are especially effective
against A549 lung cancer cells. In this biogenic synthesis, Streptomyces sp. acts as
a reducing agent for the biosynthesis of ZnO NPs (Subbaiya etal. 2017).
Antimicrobial Activity ofNP: Mechanism ofAction
Actinomycetes-mediated NP has noteworthy antimicrobial properties. The mode of
action of their microbicidal activity varies according to the type of metal nanoparticles and the specic pathogens they target. One critical mechanism involves the
physical interaction of metal nanoparticles with the cell membranes of pathogens.
This interaction impairs the integrity of the membranes, causing them to become
more permeable, which allows essential components to leak out, ultimately resulting in pathogen death (Gabrielyan and Trchounian 2019). NPs also produce reactive
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oxygen species (ROS), which can harm cellular components, such as DNA, lipids,
and proteins. Some NPs are taken up by pathogens, disrupt cellular functions, and
interact with the intracellular components of the pathogen (Cui etal. 2013). Another
signicant mechanism is the release of toxic metal ions by nanoparticles, which
bind to cellular proteins and enzymes, leading to the disruption of vital metabolic
pathways of the pathogen (Nisar etal. 2019).
S. Paul et al.
8.6 Toxicity ofMetal Nanoparticles
Nanotoxicology is a matter of growing concern because of the potential health implications of exposure to metal nanoparticles in both humans and the environment.
Exposure to nanoparticles can occur through inhalation, ingestion, or skin contact
(Yao etal. 2019). The skin serves as a critical barrier against harmful substances, but
nanoparticles can penetrate it depending on their size. Such penetration can lead to
local inammation, abnormal extracellular matrix production, and increased neovascularization (Shari etal. 2020). When ingested, nanoparticles tend to accumulate in
the liver, affecting glutathione levels, mitochondrial function and increasing reactive
oxygen species (ROS). Inhaled nanoparticles can adversely affect the respiratory
system and induce oxidative stress (Sengul and Asmatulu 2020). Some studies have
suggested that AgNPs can disrupt the integrity of the plasma membrane and disturb
mitochondrial function, ultimately leading to cell death through apoptosis. Iron
nanoparticle exposure can cause structural alterations in the root cell walls of plants.
Several variables, including size, shape, structural characteristics, dosage, dispersion, chemical composition, and capping agents, can inuence the lethal nature of
nanoparticles (Yao etal. 2019). Proper dispersion and well-dened properties can
render nanoparticles less toxic (Sengul and Asmatulu 2020). As the utilization of
nanoparticles continues to expand, concerns about their environmental impact are
also increasing, as they can disperse into water bodies, air, and soil.
8.7 Future Prospects
Although actinomycete-based metal nanoparticle production has received increasing interest of researchers worldwide in recent decades, further research is necessary to identify the putative actinomycetes. Compared to physicochemical methods,
biogenic nanosynthesis is a time-consuming procedure that takes several hours.
More extensive research is required to increase the synthesis rate. The size, shape,
and surface area of nanoparticles play a critical role in their bioactivity; however,
much research has yet to be conducted on the regulation of various biological processes that modify the size and shape of nanoparticles. Addressing concerns related
to toxicity and ensuring repeatability and biosafety are essential factors to consider
during the procedure.
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8.8 Conclusion
Actinomycetes are well known for their capability to produce a vast array of bioactive compounds and have the potential to function as nanomachines for the synthesis of a distinct range of NPs. Biogenic production of NPs by actinomycetes offers
signicant possibilities for the development of nano-antimicrobials, which could
prove to be effective substitutes and nd widespread use in various domains.
Although several studies on the biogenic synthesis of NPs from actinomycetes have
been reported, there has been growing interest in actinobacterial NP synthesis over
the past decade. However, further research is needed, with a greater focus on scaling
up processes for actinobacterial NP synthesis to meet industrial requirements.
Additionally, advancements in optimizing diverse culture conditions to enhance NP
biosynthesis efciency and provide effective control over NP size and monodispersity could pave the way for notable industrial applications.
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