Arabian Journal of Chemistry (2019) 12, 3576–3600

King Saud University

Arabian Journal of Chemistry

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

REVIEW

Biogenic synthesis of nanoparticles: A review

Deepali Sharma, Suvardhan Kanchi *, Krishna Bisetty *

Department of Chemistry, Durban University of Technology, P.O. Box 1334, Durban 4000, South Africa

Received 25 August 2015; accepted 5 November 2015

Available online 25 November 2015

KEYWORDS Abstract The nature acts like a large ‘‘bio-laboratory” comprising of plants, algae, fungi, yeast,
Nanoparticles; etc. which are composed of biomolecules. These naturally occurring biomolecules have been iden-
Green synthesis; tified to play an active role in the formation of nanoparticles with distinct shapes and sizes thereby
Algae; acting as a driving force for the designing of greener, safe and environmentally benign protocols for
Waste materials; the synthesis of nanoparticles. The present review targets the comparative biogenic synthesis and
Computational chemistry mechanisms of nanoparticles using algae and waste materials (agro waste in the presence of biomo-
lecules). The use of waste materials not only reduces the cost of synthesis but also minimizes the
need of using hazardous chemicals and stimulates ‘green synthesis’. It also focuses on the compu-
tational aspects of binding of biomolecules to nanoparticles and some of the applications of the
biosynthesized nanoparticles in biomedical, catalysis and biosensors fields.
Ó 2015 The Authors. Published by Elsevier B.V. on behalf of King Saud University. This is an open access
article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3577
2. Capping agents and their types . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3577
2.1. Biomolecules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3578
2.2. Polysaccharides. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3578
3. Understanding the role of capping agents or biomolecule binding to NPs via computational techniques . . . . . . . . . . 3578
3.1. Facet-specific binding of biomolecules for programmed biomimetic synthesis of NPs . . . . . . . . . . . . . . . . . . . 3579
4. Synthesis of NPs using algae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3580
4.1. Metallic NPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3580
4.2. Metal oxide NPs. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3582
* Corresponding authors. Tel.: +27 31 3736008/2311; fax: +27 86
6740243.
E-mail addresses: ksuvardhan@gmail.com (S. Kanchi), bisettyk@
dut.ac.za (K. Bisetty).
Peer review under responsibility of King Saud University.

Production and hosting by Elsevier

http://dx.doi.org/10.1016/j.arabjc.2015.11.002
1878-5352 Ó 2015 The Authors. Published by Elsevier B.V. on behalf of King Saud University.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Biogenic synthesis of nanoparticles: A review 3577

4.3. Mechanism of biosynthesis of NPs using Algae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3586

5. Synthesis of NPs using waste materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3586
5.1. Metallic NPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3586
5.2. Metal oxide NPs. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3589
5.3. Mechanism of waste material mediated synthesis of NPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3592
5.4. Advantage of waste material in the synthesis of NPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3592
6. Applications of biogenic synthesis of NPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3593
6.1. Biomedical applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3593
6.2. Catalytic applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3595
6.3. Biosensing applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3595
7. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3595
Future prospects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3595
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3596
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3596

1. Introduction
In the past decade there has been a marked increase in the field of fab-
rication of nanoparticles with controlled morphologies and remarkable
features making it an extensive area of research. The synthesis of
nanoparticles (NPs) with control over particle size, shape and crys-
talline nature has been one of the main objectives in chemistry that
could be used for potential applications, such as bio-medical, biosen-
sor, catalyst for bacterial biotoxin elimination and lower cost electrode
(Antonyraj et al., 2013; Frances et al., 2009; Staniland, 2007). The
nanoparticles (NPs) having at least one dimension less than 100 nm
such as nanosheets, nanotubes and nanowires have gained much atten-
tion because of their promising applications (Fu et al., 2013; Kwon and
Bard, 2012; Roy et al., 2013). Other than their unique physical and
chemical properties, NPs act as a bridge between bulk materials and
atomic or molecular structures. Therefore, they are good candidate
for applications including medical, catalysis, electrochemistry, biotech-
nology, and trace-substance detection (Luo et al., 2006; Murthy, 2007;
Wen et al., 2011; Xia et al., 2013).
Different synthetic methods have been employed for the prepara-
tion of NPs with diverse morphology and size. Although these meth-
ods have resulted in superior NPs but still a key understanding of
improved manufacturing process is required which could be exploited
at the industrial and commercial level to have better built, long lasting,
cleaner, safer and smarter products such as home appliances, commu-
nication technology, medicines, transportation, agriculture and indus-
tries. Therefore, the main focus is to design NPs using environmentally
benign approaches. These provide solutions to growing challenges
related to environmental issues.
Nature has provided ways and insight into the synthesis of
advanced nanomaterials. It has now been reported in the literature that
biological systems can act as the ‘bio-laboratory’ for the production of
pure metal and metal oxide particles at the nanometer scale using bio-
mimetic approach. Various microorganisms, such as bacteria (Shivaji
et al., 2011; Stephen and Macnaughtont, 1999), fungi (Chan and
Mat Don, 2013; Syed et al., 2013), yeast (K et al., 2011), plant extracts
(Akhtar et al., 2013) and waste materials (Kanchi et al., 2014), have
acted as eco-friendly precursors for the synthesis of NPs with potential
applications.
The biological approach which includes different types of microor-
ganisms has been used to synthesize different metallic NPs, which has
advantages over other chemical methods as this is greener, energy
saving and cost effective. The coating of biological molecules on the
surface of NPs makes them biocompatible in comparison with the
NPs prepared by chemical methods (Hakim et al., 2005; Mukherjee
et al., 2001; Tripp et al., 2002). The biocompatibility of bio-inspired
NPs offers very interesting applications in biomedicine and related
fields (Huang et al., 2015). The biogenic methods lead to the design-
ing of NPs with interesting morphologies and varied sizes (Riddin
et al., 2010; Schröfel et al., 2014); for example, Ag NPs in the size
range of 25 ± 12 nm have been prepared by exposing fungal biomass
(Verticillium) to the aqueous solution of Ag+ ions, where the NPs
were not toxic since the biomass (fungal cells) continues to grow.
The NPs were found to grow on the surface of mycelia as a result
of electrostatic interaction between the Ag+ ions and negatively
charged carboxylate groups of enzymes present in the cell wall of
the fungus (Mukherjee et al., 2001). Pt NPs of definite shape and size
have been prepared from the cell-soluble protein extract of sulfate
reducing bacteria (Riddin et al., 2010). These NPs fabricated via bio-
genic enzymatic process were superior to those synthesized via chem-
ical methods as the use of expensive chemicals was limited and they
possessed higher catalytic activity. An industrially important fungus,
Penicillium rugulosum, was used to synthesize uniform sized Au
NPs, which is easier to handle as compared to other bacteria and
yeast (Mishra et al., 2012).
Extracts from plant leaf, root, latex, seed and stem have also been
used for the synthesis of NPs as they act as stabilizing or reducing
agents. The leaf extracts of Jasminum sambac were employed to pre-
pare stable Au, Ag, Au–Ag alloy NPs (Yallappa et al., 2015). Regard-
ing the morphology control, triangular, hexagonal and spherical
shaped Au NPs were prepared using hot water, olive leaf extract at a
high reaction temperature as compared to NPs synthesized via chemi-
cal methods (Khalil et al., 2012). Iron–polyphenol (Fe–P) NPs with a
photocatalytic activity against Acid Black-194 dye were synthesized
from Australian native leaves of Eucalyptus tereticornis, Melaleuca
nesophila and Rosemarinus officinalis (Wang et al., 2014). It was noted
that the polyphenols present in the three plants reacted with ferric
chloride (FeCl3) solution to form chelated ferric–polyphenols NPs
and also led to different shapes of Fe–P NPs.
Among different biological systems used for NP synthesis, various
forms of algae are now being currently used as model systems as these
have tremendous ability of bioremediation of toxic metals thereby con-
verting them into more pliable forms. Also, these are competent in the
fabrication of diverse metal and metal oxide NPs (Patel et al., 2015).
The biosynthesis of NPs using algae and waste materials is an emerging
and upcoming research. The present review discusses the synthesis and
the comparison of the better prospects of using algae or waste material
for the efficient designing of NPs (Fig. 1).
2. Capping agents and their types
Capping agents play a very pivotal and versatile role in the NP
synthesis. NPs can be functionalized and stabilized using cap-
ping agents to impart useful properties by controlling mor-
phology, size and protecting the surface thereby preventing
aggregation. Many surfactants have been reported to be used
as capping agents for altering the desired shape and size of
3578
the NPs but these are difficult to remove and do not easily
degrade. Thus, the commercial surfactants are hazardous to
the environment (Gittins et al., 2000; Liu et al., 2005). In the
view of the limitation possessed by these chemicals, there is
an urgent need to use environment-friendly capping agents
and design green biochemical routes at laboratory and indus-
trial level for the NP synthesis. There are different types of
molecules that could act or be used as capping agents but some
of the broadly classified green capping agents have been dis-
cussed below with their potential role.
2.1. Biomolecules
The preparation of homogenous NPs using biomolecules has
recently gained interest due to their non-toxic nature and not
involving harsh synthetic procedures. Amino acids act as an
efficient reducing as well as capping agents to synthesize NPs
with unique structure. Maruyama and coworkers synthesized
Au NPs with the size range of 4–7 nm using amino acids as
capping agents. Among 20 different amino acids, they adopted
L-histidine which was found to reduce tetraauric acid (AuCl4 )
to Au NPs. The concentration of L-histidine was found to
affect the size of NPs; higher the concentration smaller the size
of NP. Moreover, the amino and carboxy groups present in the
amino acids caused the reduction of AuCl4 and coating of NP
surface (Maruyama et al., 2015). In another interesting study,
Au nanochains were prepared via facile single step within
15 min in the presence of glutamic acid and histidine amino
acids (Polavarapu and Xu, 2008). The oriented attachment
mechanism has been proposed where there is spontaneous
self-organization of the adjacent particles at a planar interface
as they share a common crystallographic orientation (Fig. 2).
The binding affinity of amino acids is found to be different
for different facets of Au crystal. The fusion of NPs along
(1 1 1) facet revealed that binding affinity of amino acids along
this facet might be weaker as compared to the other facets. The
removal of amino acid molecules from (1 1 1) facet allows the
linear aggregation of particles due to dipole–dipole interac-
tions which arise as a result of the zwitterionic nature of amino
acids (Xu et al., 2003).
Figure 1 Green synthesis of NPs using algae and waste materials.
D. Sharma et al.
2.2. Polysaccharides
Polysaccharides are a class of polymeric carbohydrate mole-
cules with repeating units of mono or disaccharides linked
together by glycosidic linkages. They act as capping agents
in the NP synthesis as they are low cost, hydrophilic, stable,
safe, biodegradable and non-toxic. The synthesis is carried
out in the presence of water as a solvent thus, eliminating
the use of toxic solvents (Akhlaghi et al., 2013; Duan et al.,
2015). One of the distinguishing features of polysaccharides
is that they sharply accelerate the kinetics of sol–gel processes
due to their catalytic effect (Boury and Plumejeau, 2015). They
not only have been found to modify the structure and mor-
phology of TiO2 but have induced a different phase where
rutile phase has been obtained in the presence of chitosan
whereas anatase in the presence of starch (Bao et al., 2013).
Dextran is a complex branched polysaccharide composed
of many glucose molecules with chains of varying lengths. It
is hydrophilic, biocompatible, non-toxic and used for coating
of many metal NPs (Virkutyte and Varma, 2011). Spherical
Au NPs of size 15 nm were synthesized in water using natural
honey which acted as reducing as well as protecting agent.
Fructose present in the honey was supposed to act as a reduc-
ing agent whereas proteins were responsible for the stabiliza-
tion of the NPs (Philip, 2009). Cheng and coworkers
synthesized Ag NPs within the size range of 2–14 nm using
aminocellulose as reducing and capping agent. Aminocellulose
is generally referred as aminodoxy derivative bearing nitrogen
functional group attached directly to the cellulose backbone. It
was predicted that at high temperature, reduction of Ag+ ions
to Ag (0) was brought by cellulose (Cheng et al., 2013). Thus,
polysaccharides have come up as one of the renewable green
alternatives for the fabrication of NPs replacing toxic chemicals
thereby saving the environment from their hazardous effects.
3. Understanding the role of capping agents or biomolecule
binding to NPs via computational techniques
With the introduction of different force field parameters and
supercomputing, computational field has given a different
Biogenic synthesis of nanoparticles: A review
Figure 2 Illustration of Au nanochain and nanowire formation through dipole–dipole interaction due to the zwitterionic nature of
amino acids. Reproduced with permission (Polavarapu and Xu, 2008).
prospective of comprehending the interaction of biomolecules
with NPs and minutely studying the mechanistic details and
supporting experiments. Different approaches such as density
functional theory (DFT), molecular dynamics (MD) simula-
tions, and docking studies are being employed to study inter-
actions which yield accurate results.
The interaction of biomolecules especially peptides on the
metal surface has been predicted to result in the stabilization
of nanostructures and hence, improvising their applicability
as sensors, biomedical devices and electronics. Phage-display
libraries have been formed to evolve peptides that can bind
specifically onto the surface of semiconductor materials
depending on the crystallographic orientation and composi-
tion. The phage display approach identifies the physical link-
age between peptide–substrate interactions. Peptides could
lead to the controlled placement and assembly of molecules
thereby broadening the scope of ’bottom-up’ approach for
the synthesis of NPs (Whaley et al., 2000). Amino acid residues
present in the biomolecules bounded to (1 1 1) facet of gold sur-
face resulting in the formation of gold nanoplates with thick-
ness of 30 nm (Shao et al., 2004). Thus, amino acids help in
the biomimetic fabrication of nanostructures.
Feng et al. explored the adsorption mechanism of amino
acids and surfactants on to the (1 1 1) surface of gold using
molecular dynamic simulation with the application of inter-
molecular potential CHARMM-METAL (Feng et al., 2011).
The molecules adsorbed onto the surface with energy between
3 and 26 kcal mol 1 and it correlated with the preferential
degree of coordination of polarizable atoms (O, N, C) to mul-
tiple epitaxial sites. The amino acids containing planar side
groups such as Arg, Trp, Gln, Met, Tyr, Asn, and PPh3 with
sp2 hybridization adsorbed strongly indicating a correlation
with molecular size and geometry. The movement of the
strongly bonded amino acids has been attributed to hopping
mechanism where surface attached guanidinium group of
Arginine moved from one favorable coordination site to
3579
another on the metal surface in the order of picoseconds
(Fig. 3).
This adsorption mechanism could give the knowledge to
control the assembly and growth of NPs.
3.1. Facet-specific binding of biomolecules for programmed
biomimetic synthesis of NPs
The bio-inspired routes have helped researchers to achieve
controllable morphology and size of NPs and come up with
promising applications. This depends on the interaction of
capping agent (biomolecules) with a particular facet of a metal
or metal oxide crystal. It is therefore, important to fully under-
stand and identify the facet specific interaction of biomolecules
with inorganic materials in order to produce NPs with hierar-
chical structures. The question still remains about the speci-
ficity of a biomolecule and its adsorption on the surface of a
particle.
Ramakrishnan et al. employed DFT method for investigat-
ing the interactions between facet specific amino acids (Phe,
Asn, Gln, Ser, Pro, Leu, Thr) and Pt (1 1 1) and Pt (1 0 0) crys-
tallographic facets. Their results indicated that the electrostatic
interactions were responsible for the binding of amino acids
onto the Pt surface. Phe, Ser and Pro amino acids preferably
adsorbed on the Pt (1 1 1) facet whereas Asn, Leu and Thr were
Pt (1 0 0) facet specific. The charge transfer and exchange pro-
cesses along with dispersive effects caused the interaction of
amino acids and Pt surface (Ramakrishnan et al., 2015).
Peptides have been found to have additional complexation
capability in the formation of ZnO crystal. The effect of ZnO
binding peptide (G-12, GLHVMHKVAPPR) and its derivative
GT-16 (GLHVMHKVAPPRGGGC) on the growth and mor-
phology of ZnO crystal was studied by Liang et al. (2011),
Sola-Rabada et al. (2015). The morphology of the ZnO crystal
was altered by G-12 and GT-16 via adsorption-growth
3580 D. Sharma et al.

Figure 3 Representative snapshot of Arg on the Au 111 surface. The guanidinium group is found most of the time in a favorable
coordination pattern with numerous epitaxial sites which leads to strong adsorption (pink highlights). Diffusion on the surface occurs by
stepwise hopping of the guanidinium group to similar epitaxial environments. In the depicted conformation, Ca and Cg point upwards
from the metal surface. Most water molecules are not shown for ease of visualization and the chloride counterion is located outside the
visible range near the metal surface. Metal atoms are shown in decreasing diameter from the top atomic layer toward adjacent atomic
layers. Reproduced with permission (Feng et al., 2011).

inhibition mechanism (Fig. 4). The aspect ratio of ZnO was

found to be reduced in the presence of G-12 and GT-16. The
inhibition of diameter growth along a-axis in the presence of
G-12 and reduction of length along c-axis in the presence of
both GT-16 and G-12 led to the following hypothesis:

(i) G-12 was able to adsorb preferentially on (0 0 0 1) plane

as well as on (1 0 1 0) plane.
(ii) Selective adsorption of GT-16 on (0 0 0 1) plane.
(iii) The selectivity of GT-16 was due to presence of GGGC
tag not present in G-12.

This gave an insight into the desired tuning of ZnO mor-

phology by the selective adsorption characteristics of the bio-
molecules onto the crystal faces.
A recent study revealed the binding behavior of two pep-
tides (EAHVMHKVAPRP, EM-12 and mutant EAHVCHK-
Figure 4 Adsorption–growth–inhibition mechanism. The length
VAPRP, EC-12) on the formation of ZnO from solution
and thickness of the growth arrows reflect the growth rate of the
(Liang et al., 2011). It was proposed that binding of ZnO did
crystals along their respective axis. Reproduced with permission
not depend on hydrophobicity but on the ZnO recognition of
(Sola-Rabada et al., 2015).
specific amino acid alignments in peptides. EM-12 suppressed
the crystal growth in the (0 0 01) direction with Met-His or
(ii) Along the c-axis the diameter of the crystal was not
His-Lys sequence of amino acid. It was also observed that with
uniform.
increase in the concentration of EM-12, there was delay in
(iii) The mushroom like morphology was obtained at higher
ZnO formation. This was attributed to the speculation that
concentration of EC-12 as the crystal planes were not
amino acids with side chain functional groups interact with
well defined.
Zn2+ via electrostatic interactions and strongly influence the
morphology. They reduce Zn2+ concentration in the solution
These observations confirm the facet specificity of biomole-
thereby delaying the ZnO formation (Gerstel et al., 2006).
cules in modulating the morphology of the NPs.
EC-12 was found to have higher Zn2+-complexation capability
due to presence of cysteine which is a strong binder and 4. Synthesis of NPs using algae
thus, retained more Zn2+ in the solution. The morphology
of the ZnO crystals was different for EC-12 as compared 4.1. Metallic NPs
to EM-12:

(i) As concentration of EC-12 was increased, the crystal The controlled growth of NPs in solution is believed to be
remained twinned. kinetically controlled process where low energy faces of any
Biogenic synthesis of nanoparticles: A review 3581

crystal results into a particular shape. The energy and growth
rate of a crystal can be controlled by the introduction of a suit-
able templating agent or a surfactant which lowers the interfa-
cial energy (Chiu et al., 2013; Xia et al., 2003). Till now,
different commercial surfactants have been used as templates
and capping agents for the synthesis of NPs with varied mor-
phologies. But the problem is the removal and complete
biodegradation of these chemicals. Nowadays, more and more
research is converged on to the green synthesis employing
environment-friendly and bioinspired approaches. By knowing
the capability of naturally occurring biomolecules to modify
the shape or size of a crystal, NPs of superior quality can be
manufactured. The use of different species of algae in the syn-
thesis of metallic NPs has stimulated the researchers to come
up with ‘nature-friendly’ methodologies.
Ag NPs have gained widespread attention due to their effi-
cient antibacterial activities. Therefore, synthesis is carried out
in the presence of microalgae where the metabolites excreted
by the algal culture cause the reduction of silver ions. The
NPs have tunable optical properties directed by the particle
size (Merin et al., 2010). The reduction of silver nitrate was
observed in the presence of seaweed Chaetomorpha linum.
The metabolites (flavonoids and terpenoids) present in the
extract were found to be effective capping, stabilizing agents
and resulted in the formation of NPs with an average size of
30 nm having potential applications in the field of medicine
(Kannan et al., 2013). Biomolecules such as polysaccharides
present in algal species also play an important role in control-
ling the size and desired shape of Ag NPs. Pterocladia, capillacae,
Jania rubins, Ulva faciata, and Colpmenia sinusa are the species
of the marine algae which are efficient in assisting the synthesis
of polydispersed and spherical Ag NPs capable of immobilizing
on cotton fabrics. Thus, they act as antimicrobial agents
(El-Rafie et al., 2013). Composite of nano Ag–CaCO3 was
manufactured via microalgae as a bio-template using efficient
and eco-friendly approach. The microalga (Chlorella sp. KR-1)
was used for the mineralization of carbon dioxide (CO2) to
form calcium carbonate (CaCO3) microspheres (Sahoo et al.,
2014). The use of alga was advantageous in the following two
ways:
(i) Nucleation and crystal growth were accelerated due to
the presence of negative charge on the surface of the cell.
(ii) Large scale synthesis at a very low cost.
The surface charge of the microalgae cells was found to be
responsible for the formation of CaCO3 particles. Due to elec-
trostatic interactions, the positively charged Ca2+ ions
agglomerated on the surface of negatively charged algal cells
and thus, were driving force for the initiation of nucleation
process. Also, the concentration of Ca2+ ions had a profound
effect on the size of microspheres. The higher the concentra-
tion, larger was the size of the CaCO3 microspheres keeping
the constant concentration of algal cells and favoring heteroge-
nous nucleation. The synthesized microspheres were used as
economical matrix for the synthesis of Ag NPs.
The composite exhibited efficient antimicrobial activity
against model bacteria such as Escherichia coli, Psychrobacter
alimenterius and Staphylococcus euroum thereby finding its
way to be commercialized as paint additives. Fig. 5 illustrates
the formation of CaCO3 microspheres and nano Ag–CaCO3
composite.
With the advent of nanotechnology, toxicity issues related
to the NPs are also of great concern. The in vitro and

Figure 5 (a) Schematic illustration of the synthetic steps for porous CaCO3 microsphere by CO2 mineralization pathway in the presence
of microalgae. Ag NPs were formed on the porous CaCO3 using AgNO3 as precursor followed by reduction. (b) Speculated mechanism for
surface-directed mineralization of CaCO3. The negatively charged microalgae surface aggregates Ca2+ ion via electrostatic interaction
prior to nucleation. Nucleation occurs on supplying CO23 from CO2 hydration resulting in the formation of nanoCaCO3. Reproduced
with permission (Sahoo et al., 2014).
3582
in vivo studies have been carried out to predict the toxicity
levels of NPs to different biological membranes and human
cells that could be administered as drugs. NP interactions are
reported to be size dependent (Kim et al., 2006). NPs with size
of ca. 10 nm have been observed to induce greater cell death
than large sized NPs (50–100 nm) (Carlson et al., 2008;
Gorth et al., 2011). The biosynthesis of metallic NPs via algae
provides a greener route with no toxicity levels. The proteins
present in the algae membranes act as better templating agents
and also stabilize the NPs thereby enhancing their possibility
to be used as nanomedicine. Marine microalgae are constantly
exposed to metal salts present in the water. Therefore, these
are widely effective in reducing metal salts to NPs. The biggest
challenge in nanochemistry is the fabrication of monodis-
persed NPs which is difficult to achieve even in the presence
of long chain surfactant molecules. Algae provide a green, fast,
cheap route and act as potential ‘nano-reserves’ for varied sus-
tainable NPs (Baker et al., 2013). Moreover, these can be easily
cultured in laboratories.
Degradation of hazardous organic chemicals and dyes is a
crucial problem. Many techniques such as activated carbon
sorption, electrocoagulation, UV degradation and redox treat-
ments are used but these are not found to be very efficient and
cost effective. Therefore, there is an urgent need of improved
ways to degrade chemicals as well as treatment of wastewater.
Ag nanocatalysts degrading methyl orange dye were synthe-
sized from Hypnea musciformis at room temperature
(Ganapathy Selvam and Sivakumar, 2015; Kumar et al.,
2011). Similarly, Ag NPs with face centered cubic crystalline
structure have been synthesized from green alga Enteromorpha
flexuosa and these demonstrated antibacterial activity against
gram negative and positive bacteria (Yousefzadi et al., 2014).
The extracellular synthesis of monodispersed Au NPs has
been achieved in a short duration from marine alga Sargassum
wightii Greville. The alga caused the reduction of auric chlo-
ride solution and the AuNPs were stable in solution which is
Figure 6 TEM images of Kf cells within silica before (a) and
after (b) gold addition. Gold colloids could be found within the
cells in the thylakoids (c), in the cell EPS (d) and in the
surrounding silica gel (e) ED pattern of dark dots with examples
of attribution to the cfc Au structure (f). Reproduced with
permission (Sicard et al., 2010).
D. Sharma et al.
important from biological prospective (Singaravelu et al.,
2007). Stoechospermum marginatum is brown algae with a high
metal binding capacity and constitutes proteins, vitamins,
amino acids, fatty acids, minerals and trace elements. Au
NPs were prepared from this algal biomass with a reaction
time of 10 min (Arockiya Aarthi Rajathi et al., 2012). Au
NPs have been reported to be intracellularly synthesized in
the suspensions encapsulated within silica gels in the presence
of Klebsormidium flaccidium algal cells giving rise to ‘‘living”
bio-hybrid material. TEM images show algal cells before and
after the addition of gold (Fig. 6). The cells are surrounded
by the colloidal silica which is not in direct contact with the
cells thereby maintaining the ability to synthesize Au NPs
intracellularly. The proximity of silica cells with cell membrane
was found to be dependent on the physiological state of algae.
This pathway proves to be one of the promising ’green’ routes
for the synthesis of NPs (Sicard et al., 2010).
The diversity in the algae species has led to their exploita-
tion and even the edible forms of algae are used in the synthesis
of metallic NPs. Au NPs have paved a way in the field of
science and technology due to their enormous vital applica-
tions as optical, electronic, catalytic, biosensors and drug car-
riers. They exhibit excellent antimicrobial and antioxidant
properties as their surface is functionalized (Ghosh et al.,
2008; Hu et al., 2006; Mikami et al., 2013; Nair and
Pradeep, 2002; Pingarrón et al., 2008; Zhao et al., 2010). The
chemical methods employ solvents which are toxic in nature
and pose a limitation for the use of NPs in the medical field.
The biomatrix of freshwater, edible red alga Lemanea fluviatilis
has been used to accomplish the synthesis of Au NPs. The pro-
teins present in the alga acted as templating as well as stabiliz-
ing agent, thereby avoiding the use of surfactants which are
difficult to remove (Sharma et al., 2014b). Prasiola crispa is
another freshwater green algae which is used for the one step
biosynthesis of Au NPs in the size range of 5–25 nm via reduc-
tion of chloroauric acid (Sharma et al., 2014a). It has been
reported that algal morphology along with high pH, metal
ion concentration and biomass concentration are important
factors for the monodispersity of the NPs. Though with time,
on exposure of NP the toxicity levels against alga increase, but
the synthesized NPs show no toxicity when tested with cell
lines of normal human cell (Parial and Pal, 2015). Thus,
algal-NPs interactions are very important considering the
effective use of NPs in drug delivery (Fig. 7).
In summary, Table 1 depicts the compositions, shape and
size of the NPs and the corresponding bio-materials adopted
as the reduction agents reported during the last decade.
4.2. Metal oxide NPs
Metal oxides are widely explored and studied class of inor-
ganic solids due to a wide variety of structures, properties
and exceptional phenomenon exhibited by their NPs. Transi-
tion metal oxides have been used in numerous industrial appli-
cations. NPs, nano-powders and nanotubes play a significant
role in industry, environmental remediation, medicine and
even in household applications. One dimensional (1D) metal
oxide nanostructures are ideal systems for exploring size and
morphology dependent applications and have become the
focus of current research efforts in the field of nanoscience
and nanotechnology. Metal oxides are commonly available
Biogenic synthesis of nanoparticles: A review 3583

Figure 7 Microphotographs of Au3+ exposed algal filaments showing changes in morphological and reproductive behavior at different
time points. (a) Akinete formation within 12 h; (b) series of akinetes at 24 h; (c) Au NP deposition started in vegetative cells and akinetes
after 24 h; (d) cell wall thickening after 48 h; (e) giant cell with complete Au NP deposition after 48 h; (f) pyknotic cell after 48 h; (g) Au
NP deposition at the periphery of cells; (h) degradation of chlorophyll and Au NP deposition within cell after 48 h; (i) purple colored
filament after 72 h. Scale bars, 20 lm (inset showing control filaments; scale bar, 40 lm). Reproduced with permission (Parial and Pal,
2015).

and present in different forms possessing special shapes, com-
position, structures, chemical and physical properties (Zhai
et al., 2009).
Different synthetic methods such as hydrothermal,
solvothermal, microwave, vapor deposition, seed mediated,
spray pyrolysis, wet-chemical have been employed for the
preparation of metal oxide NPs with diverse morphology
and size (Gotić and Musić, 2008; Hayashi and Hakuta, 2010;
Kim et al., 2003; Leonelli and Lojkowski, 2007). The reports
on the biosynthesis of metal oxide NPs are very few. The
research is now being shifted from conventional synthetic
methods to biosynthesis process where microorganism could
be employed for synthesizing NPs. Very few articles have
reported the biosynthesis approaches, especially using algae.
Copper oxide NPs have been prepared from brown alga,
Bifurcaria bifurcata extract with a dimension between 5 and
45 nm (Fig. 8). The NPs were stable in solution which is promising
for their use in biomedical applications. These NPs exhibited
antibacterial activity against bacterial strains of Enterobacter
aerogenes and Staphylococcus aureus (Abboud et al., 2014).
The extracts of green seaweed Caulerpa peltata, red Hypnea
Valencia and brown Sargassum myriocystum were also used for
the biosynthesis of ZnO NPs. The NPs with different mor-
phologies such as spherical, triangular, radial, hexagonal,
rod and rectangular were formed with size range of 76–
186 nm. The water soluble pigments present in the extract were
found to be responsible for the reduction and stabilization of
the NPs (Nagarajan and Arumugam Kuppusamy, 2013).
AFM study of ZnO NPs revealed that change in temperature
parameter remarkably affected the size and morphology of
the particles (Fig. 9).
The polysaccharides present in the aqueous extract of Sar-
gassum muticum caused the reduction of ferric chloride solu-
tion, thereby leading to the formation of ferric oxide (Fe3O4)
NPs with cubic morphology and average diameter of 18
± 4 nm (Mahdavi et al., 2013). S. muticum is algal seaweed
which is also used as functional food consisting of large quan-
tities of lipids, minerals, vitamins and many other bioactive
substances such as polysaccharides, proteins and polyphenols.
These biomolecules are effective against cancer, diabetes,
thrombosis, obesity, and other degenerative diseases
(Miyashita, 2009; Namvar et al., 2012; Nishino et al., 1999;
Perez et al., 1998) and also act as reducing as well as capping
agents. ZnO nanomaterials are considered to be biocompatible
for the medical applications. Therefore, research is carried out
to develop algae mediated one step, green and eco-friendly
approach. Recent report on the synthesis via aqueous extract
of brown alga S. muticum has indicated the formation of pure
ZnO NPs in the size range of 30–57 nm with hexagonal crystal
structure (Azizi et al., 2014). Francavilla et al., designed a fea-
sible protocol for the synthesis of ZnO NPs using Gracilaria
gracilis, an edible form of algae (Francavilla et al., 2014). This
protocol involved the solid state grinding of zinc precursors
including a biomass extracted from microalga followed by
thermal decomposition in the absence of any solvent. The syn-
thesized NPs were found to degrade phenol efficiently
3584 D. Sharma et al.

Table 1 Algae mediated synthesis of metallic NPs.

Composition of NPs Species of Algae Size (nm) Morphology Citation
Brown, Sargassum muticum Spherical

Au 5.42 ± 1.18 Namvar et al. (2015)

Tetraselmis kochinensis Spherical and triangular

Au 5–35 Senapati et al. (2012)

Brown, Ecklonia cava Spherical and triangular

Venkatesan et al.
Au 30 ± 0.25
(2014)

Caulerpa racemosa Spherical and Triangular

Kathiraven et al.
Ag 5–25
(2015)

Brown, Cystophora moniliformis Spherical

Ag 50–100 Prasad et al. (2013)

Chlamydomonas reinhardtii Round/rectangular

Ag 5–35 Barwal et al. (2011)

Biogenic synthesis of nanoparticles: A review 3585

Table 1 (continued)
Composition of NPs Species of Algae Size (nm) Morphology Citation
Chlorella vulgaris Spatial array of self assembled Structures

Annamalai and
Au 2–10
Nallamuthu, (2015)

Phaeodactylum tricornutum

Scarano and Morelli

CdS - NA
(2003)

Brown, Padina gymnospora Spherical

Au 53–67 Singh et al. (2013)

Brown, Fucus vesiculosus Spherical

Au Varied Mata et al. (2009)

Euglena gracilis Spherical

2-lines ferrihydrite
0.6–1.0 Brayner et al. (2012)
nanoparticles

NA = not available.

(Fig. 10A). ZnOAG were agar synthesized and ZnOAA alginic
acid as compared to commercial P25 Evonik. The photocat-
alytic activity was enhanced in the case of agar synthesized
(52% phenol degradation, Fig. 10B) whereas the degradation
of phenol was less than 35% in the case of ZnOAA. The contri-
bution to enhanced photocatalytic activity in this case is still
under study but the methodology could be applied widely
for the synthesis of other metal oxide NPs.
The mechanism for the formation of NPs in the presence of
algae is yet to be fully understood. The algal membranes
consist of biomolecules such as polysaccharides, proteins and
enzymes which catalyze the reduction of metal salt precursors
into metal or metal oxide NPs. Since, these are large molecules
and amphiphilic in nature, they act as surfactant molecules
which causes not only concentration buildup of the surfactant
at the surface and reduction of the surface tension, but also the
orientation of the molecule at the surface (Bianchi et al., 2006;
Liu et al., 2014). They act as capping agents and thus, reduce
the interfacial energy. The interaction of these biomolecules
with the NPs is also an important aspect under consideration.
3586
4.3. Mechanism of biosynthesis of NPs using Algae
With the availability of sophisticated biochemical techniques
and instruments, it is easy to identify the role and interaction
of a particular biomolecule such as polysaccharides, proteins,
enzymes present in the organism with NPs thereby compre-
hending the mechanism. The simple mechanism in Fig. 11
explains that enzymes and functional groups present in the cell
walls of algae form complexing agents with the precursors
thereby, causing reduction and deposition of metal/metal
oxide NPs at ambient conditions (Crookes-Goodson et al.,
2008; Gade et al., 2008). The different sources of waste mate-
rials for the green synthesis of NPs was schematically repre-
sented in Fig. 12.
Figure 8 TEM image of CuO NPs synthesized from the alga
extract. Reproduced with permission (Abboud et al., 2014).
Figure 9 AFM results of ZnO NPs 2D and 3D images. (a) Unfiltered AFM image showing topographical 2D image of ZnO NPs (b) 3D
image of synthesized (c) filtered 2D AFM image of ZnO nanoparticles (d) particle size distribution of ZnO NPs. Reproduced with
permission (Nagarajan and Arumugam Kuppusamy, 2013).
D. Sharma et al.
5. Synthesis of NPs using waste materials
5.1. Metallic NPs
The environment also acts as a ‘treasure’ of waste materials
especially food waste which can be successfully utilized for
the biosynthesis of NPs. Recent literature reports the use of
biodegradable food waste for manufacturing different NPs.
The food waste material contains different organic compounds
such as polyphenols, flavonoids, carotenoids and vitamins
(Kim et al., 2012) which act as templating agents. The various
functional groups present in these compounds cause the
reduction of metal precursors. Thus, the waste acts as a
‘biofactory’.
Au NPs have been successfully synthesized using mango
peel extract. The reaction rate was found to be faster as
compared to other plant extracts. The particles were monodis-
persed in the size range of 6.03 ± 2.77 to 18.01 ± 3.67 nm and
had no biological toxicity on the normal kidney cells (CV-1) of
African green monkeys as well as normal human fetal lung
fibroblast cells (WI-38) (Yang et al., 2014). The cytotoxicity
of the NPs was assessed by treating them with cells at different
concentrations (0, 20, 40, 80 and 160 lg mL 1) for 24 h
(Fig. 13). It was observed that Au NPs had no biological
toxicity event at higher concentration of 160 lg ml 1. The
plausible reason was the functionalization of NPs with the
organic moieties present the extract of mango peel.
Wine industry produces a lot of grape waste which is a
source of ample of organic compounds that lead to the reduc-
tion of metals to NPs. The spherical and polygonal shaped Ag
NPs with an average diameter of 25–35 nm were formed by
reduction of silver ions into NPs in the presence of extract
from grape seeds (Fig. 14A). These NPs showed effective
antibacterial activity against gram negative and gram positive
Biogenic synthesis of nanoparticles: A review 3587

Figure 10 (A) Phenol degradation efficiency (measured as the relative concentration of phenol (C/Co) over time) of ZnOAA 1: 16 alginic
acid, ZnOAG 1: 2 ex. agar and P25 Evonik, (B) Photocatalytic degradation curves of phenol for ZnOAA 1: 16 alginic acid, ZnOAG 1: 2 ex.
agar and P25 Evonik. Reproduced with permission (Francavilla et al., 2014).

Figure 11 Mechanism of biosynthesis of NPs using algae.

bacteria (Xu et al., 2015). FTIR spectrum of grape seed extract
depicted peaks at 3429.14, 2954.88, 2928.66, 2855.29, 1613.67,
1524.71, 1445.81, 1380.06, 1281.05, 1103.14, 1053.63, 820.03,
773.61 cm 1 where the peak at: 3429.14 cm 1 corresponded
to stretching vibration of O–H functional group
954.88 cm 1, 2928.66 cm 1, 2,855.29 cm 1 referred to C–H
stretching vibration mode 1613.67 cm 1, 1,524.71 cm 1
referred to vibration of cyclobenzene 1103.14 cm 1, and
1,053.63 cm 1 due to C–O vibration of phenols. The changes
in the position of absorption bands were noticed in the FTIR
spectrum of Ag NPs. There were O–H stretching vibration
shifts from 3429.14 to 3436.10 cm 1 indicating the formation
of Ag NPs (Fig. 14B).
Microwave assisted methods have been used to synthesize
Ag NPs with an average diameter of 7.36 ± 8.06 nm from
orange peel extract. The compounds present in the extract
acted as capping molecules. The less agglomeration of spheri-
cal shaped NPs was observed (Fig. 15) (Kahrilas et al., 2014).
3588 D. Sharma et al.

Figure 12 Type of waste materials employed/could be employed in the synthesis of NPs.

Figure 13 Cell viability assay of the synthesized Au NPs with different sizes (6.037 ± 2.77 nm and 18.01 ± 73.67 nm) against CV-1 and
WI-38 cells. The data are represented in the form of a bar graph and plotted using means ± S.E. of triplicate determinations. Reproduced
with permission (Yang et al., 2014).

Figure 14 (A) TEM image of Ag NPs, (B) FTIR spectra of grape seed extract and Ag NPs. Reproduced with permission (Xu et al.,
2015).
Biogenic synthesis of nanoparticles: A review
Chicken eggshell membrane (ESM) is one of the nature’s
gift which goes waste. This has been utilized for the synthesis
of fluorescent Au NPs via single step under ambient conditions
(Devi et al., 2012). ESM is double layered membrane com-
posed of water-insoluble glycoproteins such as collagen (types
I, V and X), and amino acids such as glycine alanine and uro-
nic acid (Arias et al., 1991; Wong et al., 1984). The application
of EMS as reported in the literature includes the recovery of
heavy metals, template for macroporous materials and
biosensing of enzyme immobilized Au–ESM (Lee et al.,
2009; Zheng et al., 2010). The reduction process and particle
formation were monitored by absorption and fluorescence
spectral changes in addition to change in the color. The
changes were observed in absorption spectrum after impreg-
nating EMS with 10 4 M solution of HAuCl4. A strong
Figure 15 TEM images of Ag NPs by orange peel extract at two
magnification levels. White arrows indicate the presence of a thin
organic layer surrounding the Ag NPs. Reproduced with permis-
sion (Kahrilas et al., 2014).
Figure 16 (A) UV–Vis spectra of the reaction of ESM with chloroauric acid solution (10 4 M), recorded as a function of time. Curves (1)
blank HAuCl4 (2) 1 h (3) 6 h (4) 1 day (5) 3 days (6) 6 days and (7) 7 days. (B) Photographs of (a) bare eggshell membrane and pink
colored impregnated eggshell membrane with 10 4 M solution, (b) blue colored impregnated eggshell membrane with 10 2 M metal ion
solution and (c) membranes impregnated with 10 6 (1), 10 4 (2) and 10 2 M HAuCl4 (3) solutions for 7 days. Reproduced with permission
(Devi et al., 2012).
3589
maximum was observed at 295 nm due to the initial Au3+
solution indicating the removal of Au3+ from the solution.
Also a color change was visible from white to yellow showing
the adsorption of Au3+ on the membrane surface (Fig. 16).
Annona squamosa (Annonaceae) or custard apple is an edi-
ble fruit of tropical origin widely found in America and Asia
and is reported to have medicinal, antimicrobial, insecticidal
and anti-cancerous properties (Dwivedi and Gopal, 2010;
Madhumitha et al., 2012; Mukhlesur Rahman et al., 2005;
Wagner et al., 1980). The fruit pulp is consumed but peels
are discarded. Water soluble ketone and hydroxyl groups are
present in the peel which are found to be responsible for the
reduction of silver ions and formation of Ag NPs. These
groups provide stability to NPs by forming a thin layer on
the surface (Kumar et al., 2012).
Table 2 summarizes the compositions, size, and morphol-
ogy of the NPs synthesized using waste materials.
5.2. Metal oxide NPs
As compared to metallic NPs, synthesis of metal oxide NPs
using agro and food waste is yet to be explored widely. Thus,
this opens up an upcoming area of research where eco-friendly
and fast methods can be developed thereby achieving the
potential applications of the metal oxide NPs. The tea waste
template has been used for the fabrication of magnetic iron
oxide (Fe3O4) NPs in the size range of 5–25 nm with cuboid/
pyramid morphology. These NPs proved to be very efficient
in removing arsenic metal from water and could be used up
to five adsorption cycles (Lunge et al., 2014). Another material
which is considered as garbage is egg-shells from the baking,
food processing and baking industry. This is considered to
have no food value, but it has been used for the synthesis of
hydroxyapatite NPs. Eggshells contain calcium carbonate
(94%), magnesium carbonate (1%), calcium phosphate (1%)
and organic matter (4%). The high calcium content present
assists in the formation of hydroxyapatite NPs (Rivera et al.,
3590 D. Sharma et al.

Table 2 Waste material mediated synthesis of metallic NPs.

Composition of NPs Waste material Size Morphology Citation
(nm)
Cellulose Cotton fibers 40–90 Spherical Fattahi Meyabadi et al.
(2014)

Silicon carbide Electronic compact disks char 40–90 Spherical Rajarao et al. (2014)

Ag Satsuma mandarin(Citrus unshiu) 5–20 Spherical Basavegowda and Rok

peel extract Lee (2013)

Ag Industrial milk waste – Nanorods Sivakumar et al. (2013)

Fe Citrine juices 3–300 Spherical, cylindrical, Machado et al. (2014)

irregular

Au Grape skin, stalk and seeds 20–25 Quasi-spherical Krishnaswamy et al.

(2014)
Biogenic synthesis of nanoparticles: A review 3591

Table 2 (continued)
Composition of NPs Waste material Size Morphology Citation
(nm)
Au Rice bran 50–100 Spherical Malhotra et al. (2014)

Pd Watermelon rind 96 Spherical Lakshmipathy et al. (2015)

N-CNTs (Nitrogen doped carbon Poultry chicken Feather – Gao et al. (2014)
nanotubes)

Figure 17 (A) SEM image of Mn3O4 NPs (B) cyclic voltammograms of Mn3O4 (C) cycle performance during 2000 cycles at a current
density of 0.3 A g 1 (the inset shows charge–discharge curves for the first (left) and last (right) five cycles). Reproduced with permission
(Yan et al., 2014).
3592
Figure 18 Mechanism of formation of NPs in the presence of waste materials.
1999; Wu et al., 2013). Banana is a favorite food worldwide
and according to statistics which are not complete, more than
100 million tons of bananas is consumed worldwide every year
(Venkateswarlu et al., 2013) and the peel is dumped as gar-
bage. This is composed of biopolymers such as cellulose, hemi-
celluloses, pectin, lignin and proteins and could be efficiently
used for the synthesis of NPs. Hydroxyapatite NPs have been
synthesized using banana peel. Pectin present in the peel plays
a major role in the surface modification of the NPs (Chanakya
et al., 2009; Happi Emaga et al., 2008; Morra et al., 2004;
Rodrı́guez-Ambriz et al., 2008). The as-synthesized NPs exhib-
ited antibacterial activity against gram positive and negative
bacteria (Gopi et al., 2014). Banana peel extract has also been
used successfully to synthesize Mn3O4 NPs having super-
capacitive properties (Yan et al., 2014). Mn3O4 is one of most
stable oxides of manganese which possesses broad range of
interesting properties ranging from catalysis to high density
magnetic storage medium (Wang et al., 2009). SEM image
depicts the agglomerated particles with an average diameter
of 20–50 nm (Fig. 17A). Fig. 17B represents cyclic voltammet-
ric (CV) profiles which indicate high electrochemical reversibil-
ity of the as synthesized NPs. 93% of specific capacitance (SC)
was maintained after 2000 cycle numbers of charge–discharge
process thereby confirming the superior stability of Mn3O4
electrode (Fig. 17C).
5.3. Mechanism of waste material mediated synthesis of NPs
The cell walls of food waste materials contain cellulose, hemi-
celluloses, pectins, lignins, proteins and biodegradable polysac-
charides. The waste materials also contain phytochemicals
D. Sharma et al.
such as polyphenols, carotenoids, flavonoids, dietary fibers
and essential oils. These act as templating agents in the synthe-
sis of NPs thereby determining their morphology and size. The
biomolecules cause the reduction of metal salts into metal or
metal oxide NPs (Heim et al., 2002). The plausible mechanism
for the synthesis of NPs using waste materials has been dia-
grammatically shown in Fig. 18.
5.4. Advantage of waste material in the synthesis of NPs
Green synthesis has led to the fabrication of many inorganic
NPs especially, metal NPs. Although different species of algae
have been used for NP synthesis waste materials (food and
agro waste) have been found to be best suited for preparation
of NPs of diverse morphologies and sizes. The different biomo-
lecules present in the waste act as templating agents and thus,
lead to the fast, completely green and cost effective approach.
This waste is easily available and does not require rigorous
processing methods. These can be directly used in the synthesis
of NPs and also pave the way for the management of the
waste. Whereas, the preparation of algae culture requires time
and cost, there is a limitation of maintaining the cultures over
a period of time. The toxicity factors of some of the species of
the algae are also to be considered before using them for the
synthesis of NPs as certain compounds present could induce
toxicity to the NPs and thus limit their use for biomedical
applications. Moreover, limited morphologies could be
obtained via algae mediated synthesis of NPs as can be seen
in Tables 1 and 2. Thus, the main challenges related to algae
mediated synthesis of NPs are as follows (Seabra and Durán,
2015):
Biogenic synthesis of nanoparticles: A review 3593

(i) Reproducibility of the methods needs to be improved.
(ii) Limitation of scaling up of the synthesis methods.
(iii) Complete elucidation of mechanism of formation of
NPs.
(iv) Size control and monodispersity of NPs.
6. Applications of biogenic synthesis of NPs
The applications of biosynthesized NPs range from biomedical
to photocatalytic and sensors. The properties of these NPs are
different from the NPs synthesized via other conventional and
chemical methods since, no capping agents or surfactants are
involved. Thus, NPs synthesized using algae and waste materi-
als exhibit a broad new spectrum of potential applications.

6.1. Biomedical applications

Extensive research is going on and vast literature is available

Figure 19 TEM images of (A) B. subtilis (B) S. aureus cells
treated with S. platensis protein protected Au NPs in agar
medium for 1 h. Reproduced with permission (Uma Suganya
et al., 2015).
on the antimicrobial activity of NPs. Ag NPs have attracted
much attention as an efficient antimicrobial and biocompatible
agent. These can effectively bind with the cell wall which is
required for better antimicrobial activity (Eckhardt et al.,
2013). Ag NPs possess higher antibacterial activity against
E. coli than S. aureus due to the structural difference of the cell
wall (Feng et al., 2000; Jung et al., 2008).
Being super paramagnetic in nature, iron and iron oxide
NPs find extensive usage in biomedical applications such as
cell labeling, tissue repair, magnetic resonance imaging
(MRI), and drug delivery (Catherine and Adam, 2003;
Pankhurst et al., 2003). Au NPs have proved to be important
tool in many potential biomedical applications including an
emerging alternative for life-threatening diseases and also have
been used in DNA modeling (Gupta and Gupta, 2005; Khan
et al., 2013). Au NPs with different sizes display optical prop-
erties necessary for biosensor applications, especially in cancer
nanotechnology. PEG coated Au NPs maximize the tumor
damage as compared to Tumor necrosis factor-alpha (TNF-a),
a cytokine which has anticancer efficacy, but limited thera-
peutic applications (Cai et al., 2008; van Horssen et al.,
2006; Visaria et al., 2006).

Figure 20 Representation of receptor-mediated uptake. This is the specific biological mechanism for particles interacting with the
surface membrane and undergoing cellular uptake. The intrinsic NP characteristics that promote surface binding (roughness,
hydrophobicity, cation charge) generally lead to nonspecific binding forces (marked by asterisks) that promote cellular uptake. In contrast,
specific receptor–ligand interactions generally lead to endocytic uptake. A combination of nonspecific binding forces on the surface of
spiked particles can lead to direct penetration of the surface membrane without the need to involve endocytic compartments. Reproduced
with permission (Nel et al., 2009).
3594 D. Sharma et al.

Au NPs exhibit high antibacterial activity due to their
small size and high surface area. These NPs suppress the res-
piratory chain enzymes which are vital for the cell wall syn-
thesis of bacteria thereby leading to death or static growth.
Spherical Au NPs synthesized from protein extract of blue
green alga, S. platensis have been reported to show inhibitory
action against B. subtilis and S. aureu. Since, gram positive
bacteria have thick peptidoglycan layer, NPs adhere to the
membrane and break the bonds thereby entering inside the
microorganism (Uma Suganya et al., 2015). Fig. 19 depicts
the damage caused to bacterial membrane on treatment with
Au NPs.
Thus, functionalized NPs could be modified for use in
advanced medical applications with greener methods.
Banana peels rich in lignin, cellulose, pectins and hemicellu-
lose act an excellent template for the synthesis of NPs. The peel
extract has been used to synthesize Ag NPs which showed
efficacious antimicrobial activity against pathogenic bacteria

Figure 21 (A) Fe3O4 NPs coated with SBO (B) CV removal obtained with NP/0.5 at different pHs. [CV]0 = 10 mg L 1; [NP/
0.5]0 = 150 mg L 1. Reproduced with permission (Magnacca et al., 2014).

Figure 22 Palladium NP synthesis by photosynthetic green microalgae, and their uptake on an electrospun chitosan mat for use as a
catalyst in Mizoroki–Heck reactions. The left stage shows a combination of mechanisms taking place within the photosynthetic organisms,
resulting in the production of reducing agents. (ADP: adenosine diphosphate, ATP: adenosine triphosphate, Fd: ferredoxin, NADP+:
oxidized form of nicotinamide adenine dinucleotide phosphate, NADPH: reduced form of nicotinamide adenine dinucleotide phosphate,
PGA: phosphoglycolic acid, RuBisCO: rubilose biphosphate carboxylase). NADPH is likely one of the main reducing agents for the
reduction of Na2[PdCl4], which is partially oxidized as a result of aerobic culture conditions. Reproduced with permission (Eroglu et al.,
2013).
Biogenic synthesis of nanoparticles: A review
(B. subtilis, S. aureus, Pseudomonas aeruginosa, and E. coli)
and yeast (Candida albicans) (Ibrahim, 2015).
The mechanism for the interaction of NPs with the specific
membrane has been illustrated in Fig. 20. As soon as NP
comes in contact with the membrane of pathogenic microor-
ganisms, there is dissolution and release of metal cations which
inhibit respiratory enzymes and ATP production. There is
reactive oxygen species (ROS) production which disrupts
membrane integrity and other transport processes (Nel et al.,
2009).
6.2. Catalytic applications
Biosynthesized NPs exhibit interesting size dependent catalytic
properties due to high surface-to-area volume ratio. Pd NPs
synthesized using soya leaf extract caused the degradation of
azo dyes (Petla et al., 2012). Fe3O4 NPs coated with soluble
bio-based products (SBO) efficiently adsorbed crystal violet
(CV) dye used as a model pollutant (Fig. 21A). Thus, these
NPs could be used for the removal of pollutants in the water
(Magnacca et al., 2014). The effect of pH on the removal of
CV dye with NPs was studied by carrying out sorption
experiments. It was observed that as pH was increased the
% removal of dye also increased (Fig. 21B).
Ag NPs with an average diameter of 2.46 nm were prepared
from eggshell membrane (ESM) which is considered as food
waste. Ag NPs possessed good catalytic activity for the reduc-
tion of 4-nitrophenol and could be used up to eight cycles
because of high stability (Liang et al., 2014). ESM fibers were
treated with Procyanidin (Pro) extracted from grape seeds and
skin so that they could act as reductant and stabilizer. Plant
extracts are one of the agricultural wastes that has become a
new face in the green synthesis of NPs. Prunus domestica
(plum) fruit extract has been used for the synthesis of spherical
Au NPs with an average diameter of 4–38 nm which caused the
reduction of toxic pollutant 4-nitrophenol to 4-aminophenol
(Dauthal and Mukhopadhyay, 2012).
Spherical Pd NPs synthesized within green microalgae
(Chlorella vulgaris) acted as a catalyst for Mizoroki–Heck
cross-coupling reaction (Fig. 22). They were synthesized via
photosynthetic reaction and thus provide a green route of syn-
thesis for other NPs (Eroglu et al., 2013).
Pd NPs have also been reported to synthesize directly on
alginic acid (AA) and seaweed (Laminaria digitata, a brown
alga with the common name Oarweed). The reaction of
iodobenzene and methyl acrylate was used to determine the
catalytic activity. It was noted that 75% yield was obtained
after 20 min in the presence of Pd NPs with a reusability of
2–3 times (Parker et al., 2015).
6.3. Biosensing applications
The biosensing applications of algae and waste mediated syn-
thesized NPs are under study and would be preferred over
commercially synthesized NPs. Here, in brief, biosensing abil-
ity of NPs synthesized from other sources has been discussed
which would make NPs derived from algae and waste materi-
als a better choice.
Biosynthesized Au NPs have proved to be very important
tool for hormone (HCG) detection in pregnant women urine
3595
sample (Kuppusamy et al., 2014). Adrenaline acts as a drug
which is widely used in the treatment of allergies, heart attack,
asthma and cardiac surgery. Therefore, detection of adrenalin
is becoming an active area of research from medical point of
view. Pt NPs have been acted as a novel biosensor with high
sensitivity for the determination of adrenaline (Brondani
et al., 2009). Nanoscale Au–Ag alloy prepared via chloroplasts
exhibited high electrocatalytic activity for 2-butanone at room
temperature thereby providing a platform for the development
of biosensor capable of detecting cancer at early stages (Zhang
et al., 2012).
7. Conclusions
The use of different types of algae in the synthesis of NPs has encour-
aged the designing of simple, green, cost and time effective approaches
thereby, minimizing the use of chemicals and solvents. The polysaccha-
rides, proteins and lipids present in the algal membranes act as capping
agents and thus limit the use of non-biodegradable commercial surfac-
tants which are difficult to remove after the synthesis of NPs. But the
limitation with the use of algae is that, not all the species can be
exploited for the synthesis as some of them contain toxic compounds
and moreover, the mechanism for synthesis has not been fully explored
yet. This limitation has led to the way for the use of waste materials in
the synthesis process of NPs. The waste material, especially fruit waste
is easily available and does not require pre-conditioning of the materi-
als. The method of synthesis is very simple, requiring less time and
energy and predictable mechanisms. This opens up an opportunity
for the use of biodegradable materials especially in the synthesis of
metal oxide NPs.
Future prospects
The biosynthesis of NPs using waste materials will help
researchers not only to design safer nanomaterials but also
to promote the understanding of health and safety considera-
tions of NPs. Useful materials can be produced easily even
at reasonable scale because the biomaterial based routes
eliminate the need to use toxic chemicals. Considerable efforts
are devoted on different capping agents such as biomolecules
and polysaccharides which can act as both chelating/reducing
and capping agents for the synthesis of NPs. Therefore, the
resulting particles are protected from further reactions and
aggregation, which increases their stability and longevity.
Greener methods that have been used in NP synthesis are gen-
erally single-pot reactions, without the use of additional sur-
factants, capping agents and templates. Survey of literature
reveals that these investigations have been carried out at labo-
ratory scale, whereas there are no reports on ‘pilot plant’ syn-
thesis of NPs using waste materials and algae. We believe that
there are good opportunities for developing industrial scale
production, where NPs have important applications. Overall,
the use of algae and waste materials for green synthesis of
NPs/nanomaterials is an emerging and exciting area of nan-
otechnology and may have significant impact on further
advances in nanoscience. Moreover, with the availability of
computational techniques, area of green synthesis and devel-
opment of nanomaterials will be broadened that could be used
in the field of medicine as ‘nanodrugs’ in the near future. Also,
these NPs would provide a potential solution for the present
energy crisis by finding their use as energy driven devices.
3596
Acknowledgments
Our grateful acknowledgement goes to the Durban University
of Technology and National Research Foundation of South
Africa for the financial support.
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