J Nanopart Res (2008) 10:507–517
DOI 10.1007/s11051-007-9275-x
TECHNOLOGY AND APPLICATIONS
Biosynthesis of nanoparticles: technological concepts
and future applications
Prashant Mohanpuria Æ Nisha K. Rana Æ
Sudesh Kumar Yadav
Received: 14 March 2007 / Accepted: 6 July 2007 / Published online: 3 August 2007
Ó Springer Science+Business Media B.V. 2007
Abstract Nanotechnology involves the production,
manipulation and use of materials ranging in size
from less than a micron to that of individual atoms.
Although nanomaterials may be synthesized using
chemical approaches, it is now possible to include the
use of biological materials. In this review, we
critically assess the role of microorganisms and
plants in the synthesis of nanoparticles.
Keywords Biologicals
Nanoparticles biosynthesis

Nanoparticles applications
Nanotechnology
Plants

Microorganisms
Introduction
Synthesis of nanoparticles using biological entities
has great interest due to their unusual optical
(Krolikowska et al. 2003), chemical (Kumar et al.
2003), photoelectrochemical (Chandrasekharan and
Kamat 2000), and electronic (Peto et al. 2002)
properties. A wide variety of physical, chemical and
biological processes results in the synthesis of
nanoparticles, some of these are novel and others
are quite common. Nature has devised various
P. Mohanpuria
N. K. Rana
S. K. Yadav (&)
Biotechnology Division, Institute of Himalayan
Bioresource Technology, CSIR, Palampur 176061, India
e-mail: skyt@rediffmail.com; sudeshkumar@ihbt.res.in
processes for the synthesis of nano- and micro-length
scaled inorganic materials which have contributed to
the development of relatively new and largely
unexplored area of research based on the biosynthesis
of nanomaterials (Sastry et al. 2004). The synthesis
and assembly of nanoparticles would benefit from the
development of clean, nontoxic and environmentally
acceptable ‘‘green chemistry’’ procedures, probably
involving organisms ranging from bacteria to fungi
and even plants (Bhattacharya and Rajinder 2005;
Sastry et al. 2004). Hence, both unicellular and
multicellular organisms are known to produce inor-
ganic materials either intra-or extracellularly (Mann
1996).
The Verticillium sp. fungal biomass when exposed
to aqueous AgNO3 solution resulted in the intracel-
lular formation of silver nanoparticles, while Fusa-
rium oxysporum biomass resulted in the extracellular
silver nanoparticles (Senapati et al. 2004). The use of
microorganisms such as bacteria, yeast, fungi and
actinomycetes has been described for the formation
of nanoparticles and their applications (Sastry et al.
2003; Mandal et al. 2006; Gericke and Pinches
2006a). The rate of intracellular particle formation
and therefore the size of the nanoparticles could, to
an extent, be manipulated by controlling parameters
such as pH, temperature, substrate concentration and
exposure time to substrate (Gericke and Pinches
2006a). Efforts have also been made to manipulate
the shape and size of gold nanoparticles produced
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extracellularly by microorganisms through altering
key growth parameters (Gericke and Pinches 2006b).
Efforts are also being given for the synthesis of
nanoparticles of different chemical composition,
sizes and controlled monodispersity. For example
synthesis of biominerals which are composite mate-
rials and consist of an inorganic component and a
special organic matrix (proteins, lipids, or polysac-
charides) that controls the morphology of the inor-
ganic compound. We reviewed here on the synthesis
of nanoparticles using various living organisms.
Use of organisms in the synthesis of nanoparticles
Here, we summarize some of the organisms used in
the biosynthesis of nanomaterials and describe the
properties that should be inherent for the production
of nanoparticles of desired characteristics. Some
organisms which have been used for the production
of nanoparticles are summarized in Table 1.
Use of bacteria
Pseudomonas stutzeri AG259 isolated from silver
mines has been shown to produce silver nanoparticles
(Joerger et al. 2000; Klaus et al. 1999). The synthesis
of magnetic nanoparticles has been reported by using
magnetotactic bacteria (Roh et al. 2001). Magneto-
tactic bacteria such as Magnetospirillum magneticum
produce two types of particles; some produce mag-
netic (Fe3O4) nanoparticles in chains and some
produce greigite (Fe3S4) nanoparticles, while some
other produce both types of nanoparticles. Similarly
in the presence of exogenous electron donor, sul-
phate-reducing bacterium Desulfovibrio desulfuri-
cans NCIMB 8307 has been shown to be
synthesizing palladium nanoparticles (Yong et al.
2002). Nair and Pradeep (2002) reported that com-
mon Lactobacillus strains found in buttermilk
assisted the growth of microscopic gold, silver, and
gold-silver alloy crystals of well-defined morphology.
Recently, bacterial cell supernatant of Pseudomonas
aeruginosa was used for the reduction of gold ions
resulting in extracellular biosynthesis of gold nano-
particles (Husseiny et al. 2007). This would help in
understanding the biochemical and molecular mech-
anism of nanoparticles synthesis. The cell filtrate has
helped in achieving better control over size and
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J Nanopart Res (2008) 10:507–517
polydispersity of nanoparticles. Further, this has
documented that extracellular synthesis of nanopar-
ticles using cell filtrate could be beneficial over
intracellular synthesis.
Morphological control over the shape of gold
nanoparticles has been achieved by using Plectonema
boryanum UTEX 485, a filamentous cyanobacterium.
When it was reacted with aqueous Au(S2O3)32 and
AuCl4 solutions at 25–100 °C for up to 1 month and
at 200 °C for 1 day resulted in the precipitation of
cubic gold nanoparticles and octahedral gold plate-
lets, respectively (Lengke et al. 2006a). The mech-
anisms of gold bioaccumulation by cyanobacteria
(Plectonema boryanum UTEX 485) from gold(III)-
chloride solutions have documented that interaction
of cyanobacteria with aqueous gold(III)-chloride
initially promoted the precipitation of nanoparticles
of amorphous gold(I)-sulfide at the cell walls, and
finally deposited metallic gold in the form of
octahedral (III) platelets near cell surfaces and in
solutions (Lengke et al. 2006b). Adding further to the
mechanism, a sulfate-reducing bacterial enrichment
was used to destabilize gold(I)-thiosulfate complex to
elemental gold and proposed that this could occur by
three possible mechanisms involving iron sulfide,
localized reducing conditions, and metabolism
(Lengke and Southam 2006).
Use of actinomycetes
It has been observed that the extremophilic actino-
mycete, Thermomonospora sp. when exposed to gold
ions reduced the metal ions extracellularly, yielding
gold nanoparticles with a much improved polydis-
persity (Sastry et al. 2003). However, in an effort
towards elucidating mechanism or conditions favour-
ing the formation of nanoparticles with desired
features, Ahmad et al. (2003a) carried out the reduc-
tion of AuCl4 ions by using an extremophilic
Thermomonospora sp. biomass that has resulted in
efficient synthesis of monodisperse gold nanoparti-
cles. The reduction of metal ions and stabilization of
the gold nanoparticles were believed to occur by an
enzymatic process (Ahmad et al. 2003a, b). They
compared their earlier work of synthesis of polydis-
perse gold nanoparticles by using Fusarium oxyspo-
rum to this study and concluded that the
monodisperse gold nanoparticles synthesis could be
due to extreme biological conditions such as alkaline
J Nanopart Res (2008) 10:507–517 509

Table 1 Use of various biological entities in the production of nanoparticles

Biological entity Nanoparticles produced Size Extracellular/ Reference
intracellular

Avena sativa (Plant) Au 5–20 nm (at pH 3 and 4), Intracellular Armendariz et al. (2004)
25–85 nm (at pH 2)
Azadirachta indica (Plant) Ag, Au and Ag/Au 50–100 nm Extracellular Shankar et al. (2004)
bimetallic
Aloe vera (Plant) Ag 15.2 ± 4.2 nm Chandran et al. (2006)
Alfalfa (Plant) Ti/Ni bimetallic 1–4 nm Schabes-Retchkiman
et al. (2006)
Aspergillus fumigatus (Fungus) Ag 5–25 nm Extracellular Bhainsa and D’ Souza
(2006)
Colletotrichum sp. (Fungus) Au 20–40 nm Extracellular Shankar et al. (2003a)
Candida glabrata (Yeast) CdS 20 Å Intracellular Dameron et al. (1989)
Desulfovibrio desulfuricans Palladium – – Yong et al. (2002)
(Bacterium)
Emblica Officinalis (Plant) Ag and Au (10–20 nm) and (15– Extracellular Ankamwar et al. (2005a)
25 nm )
Fusarium oxysporum and Magnetite 20–50 nm Extracellular Bharde et al. (2006)
Verticillium sp. (Fungi)
Fusarium oxysporium (Fungus) Ag 5–15 nm Extracellular Ahmad et al. (2003)
Fusarium oxysporium (Fungus) Au 20–40 nm Extracellular Mukherjee et al. (2002)
Fusarium oxysporum (Fungus) Zirconia 3–11 nm Extracellular Bansal et al. (2004)
Fusarium oxysporum (Fungus) CdS 5–20 nm Extracellular Ahmad et al. (2002)
Fusarium oxysporum (Fungus) Barium titanate 4–5 nm Extracellular Bansal et al., 2006
MKY3 (Yeast) Ag 2–5 nm Extracellular Kowshik et al. (2003)
Magnetotactic bacteria Magnetic (Fe3O4), – – Roh et al. (2001)
greigite (Fe3S4)
Rhodococcus Au 5–15 nm Intracellular Ahmad et al. (2003a)
sp.(Actinomycete)
Pseudomonas aeruginosa Au 15–30 nm Extracellular Husseiny et al. (2007)
(Bacterium)
Pelargonium graveolens (Plant) Ag 16–40 nm Extracellular Shankar et al. (2003)
Schizosaccharomyces pombe CdS 1–1.5 nm Intracellular Kowshik et al. (2002)
(Yeast)
Pseudomonas stutzeri Ag Up to 200 nm Joerger et al. (2000);
(Bacterium) Klaus et al. (1999)
Schizosaccharomyces pombe CdS 20 Å Intracellular Dameron et al. (1989)
(Yeast)
Trichothecium sp. (Fungus) Au – Extracellular Ahmad et al. (2005)
and
Intracellular
Thermomonospora sp. Au 8 nm Extracellular Ahmad et al. (2003b)
(Actinomycetes)
Verticillium (Fungi) Au 20 nm Intracellular Mukherjee et al. (2001a)
Verticillium (Fungi) Ag 25 ± 12 nm Intracellular Mukherjee et al. (2001);
Senapati et al. (2004)
P. jadinii (Yeast) Au Few to 100 nm Intracellular Gericke and Pinches
(2006b)
V. luteoalbum (Fungus) Au Few to 100 nm Intracellular Gericke and Pinches
(2006b)

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Table 1 continued
Biological entity Nanoparticles produced
Thermomonospora sp. Au
(Actinomycete)
Plectonema boryanumUTEX Octahedral Au platlets
485 (Cyanobacterium)
Cinnamomum camphora Au and Ag
(Plant)
Nitrate reductases ( from Ag
Fusarium oxysporum, a
Fungus)
Fusarium oxysporum,( Fungus) CdSe quantum dots
Fusarium oxysporum,( Fungus) Silica and Titanium
particles (SiF26 and
TiF26 )
Tamarind Leaf Extract (Plant) Au nanotriangles
and slightly elevated temperature conditions used for
the synthesis of nanoparticles. Based on the hypoth-
esis alkalotolerant Rhodococcus sp. has been used for
intracellular synthesis of good quality monodisperse
gold nanoparticles. They observed that the concen-
tration of nanoparticles were more on the cytoplasmic
membrane than on the cell wall. This could be due to
reduction of the metal ions by enzymes present in the
cell wall and on the cytoplasmic membrane but not in
the cytosol. These metal ions were not toxic to the
cells, which are producing them, and were continued
to multiply even after the biosynthesis of gold
nanoparticles (Ahmad et al. 2003a).
Use of yeast
The Candida glabrata and Schizosaccharomyces
pombe were used for the first time in the biosynthesis
of cadmium sulphide (CdS) nanocrystals. These
nanocrystals were produced using cadmium salts
and are now used in quantum semiconductor crystal-
lites (Dameron et al. 1989). Further experiments
have been conducted to improve the quantity of
semiconductor CdS nanocrystals production that was
achieved by using Schizosaccharomyces pombe cells.
When these cells were incubated with 1mM Cd
during their mid-log phase of growth, maximal
nanocrystals were obtained (Kowshik et al. 2002).
This study has suggested that the formation of CdS
nanocrystals was dependent on the growth phase of
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Size Extracellular/ Reference
intracellular
– Extracellular Sastry et al. (2003)
6 lM to 10 nm At the cell wall Lengke et al. (2006a, b)
55–80 nm Extracellular Huang et al. (2007)
10–25 nm Extracellular Kumar et al. (2007a)
– Extracellular Kumar et al. (2007b)
5–15 nm Extracellular Bansal et al. (2005)
20–40 nm Extracellular Ankamwar et al. (2005b)
yeast. When Cd was added during stationary phase,
its uptake as well as production of CdS nanocrystals
was decreased or resulted in no CdS formation. Upon
adding Cd during early exponential phase of yeast
growth, CdS nanocrystals were formed but this time
it was affecting the cellular metabolism of the yeast
and resulted in efflux of Cd from the cells (Williams
et al. 1996). The possible mechanism of decrease in
CdS nanocrystals formation could be like this: upon
exposure to Cd as a stress, a series of biochemical
reactions were triggered to overcome the toxic effects
of this metal. Firstly, an enzyme phytochelatin
synthase was activated to synthesize phytochelatins
(PC) that chelated the cytoplasmic Cd to form a low
molecular weight PC-Cd complex and ultimately
transport them across the vacuolar membrane by an
ATP-binding cassette-type vacuolar membrane pro-
tein (HMT1). In addition to Cd, sulphide could also
be added to this complex in the membrane and that
results in formation of high molecular weight PC-
CdS2 complex that also allow them to sequestered
into vacuole (Ortiz et al. 1995).
Conditions have also been standardized for the
synthesis of large quantities of silver nanoparticles by
using silver-tolerant yeast strain MKY3. The proce-
dure for separation of these silver particles has also
been documented that was based on differential
thawing of the samples (Kowshik et al. 2003).
Recently, yeast strains have been identified for their
ability to produce gold nanoparticles, whereby con-
J Nanopart Res (2008) 10:507–517
trolling growth and other cellular activities controlled
size and shape of the nanoparticles was achieved
(Gericke and Pinches 2006a).
Use of fungi
In addition to good monodispersity, nanoparticles
with well defined dimensions can be obtained by
using fungi. This has been shown with an experiment
where bioreduction of aqueous AuCl4 ions was
carried out using the fungus Verticillium sp. that led
to the formation of gold nanoparticles with fairly
well-defined dimensions and good monodispersity
(Mukherjee et al. 2001a). These results have docu-
mented that the trapping of AuCl4 ions on the surface
of fungal cells could occur by electrostatic interaction
with positively charged groups (such as, lysine
residues) in enzymes that are present in the cell wall
of the mycelia. Here then gold ions were reduced by
enzymes within the cell wall leading to aggregation
of metal atoms and formation of gold nanoparticles.
However, they could not find the exact mechanism of
formation of gold nanoparticles. It can be concluded
from their study that compared to bacteria, fungi
could be a source for large amount production of
nanoparticles. Since fungi are known to secrete much
higher amounts of proteins, thus might have signif-
icantly higher productivity of nanoparticles in bio-
synthetic approach. Towards elucidating mechanism
of nanoparticles formation, an in vitro approach was
followed where species specific NADH dependent
reductase, released by the Fusarium oxysporum, were
successfully used to carry out the reduction of AuCl4
ions to gold nanoparticles. This has first time opened
up a novel fungal/enzyme-based in vitro approach for
nanomaterials synthesis (Mukherjee et al. 2002).
Based on properties of Fusarium oxysporum, it was
also used in the formation of extremely stable silver
hydrosol (Ahmad et al. 2003a). The acidophilic
fungus Verticillium sp. has capability of producing
gold as well as silver nanoparticles upon their
incubation with Ag+ and AuCl4 ions (Sastry et al.
2003). However, a novel biological method for the
intra- and extra-cellular synthesis of silver nanopar-
ticles using the fungi, Verticillium and Fusarium
oxysporum respectively has been documented. This
has opened up an exciting possibility wherein the
nanoparticles may be entrapped in the biomass in the
form of a film or produced in solution, both having
511
interesting commercial potential (Senapati et al.
2004). The fungus, Aspergillus flavus also resulted
in the accumulation of silver nanoparticles on the
surface of its cell wall when incubated with silver
nitrate solution (Vigneshwaran et al. 2007).
Extracellularly produced nanoparticles were stabi-
lized by the proteins and reducing agents secreted by
the fungus. A minimum of four high molecular
weight proteins released by the fungal biomass have
been found in association with nanoparticles. One of
these was strain specific NADH-dependent reductase.
However, emission band produced by fluorescence
spectra indicate the native form of these proteins
present in the solution as well as bound to the
surfaces of nanoparticles (Macdonald and Smith
1996; Kumar and McLendon 1997). Further, the
reduction of metal ions and surface binding of the
proteins to the nanoparticles did not compromise the
tertiary structure of the proteins. Endophytic fungus
collitotrichum sp. growing in the leaves of geranium
was used for the synthesis of stable and various
shaped gold nanoparticles. Reducing agent in this
fungus were also polypeptides/enzymes (Shankar
et al. 2003a). Instead of fungi culture, isolated
proteins from them have been used successfully in
nanoparticles production. Nanocrystalline zirconia
was produced at room temperature by the cationic
proteins. These proteins were similar in nature to
silicatein, secreted by the Fusarium oxysporum, and
were capable of hydrolyzing aqueous ZrF26 ions
extracellularly (Bansal et al. 2004).
Growth conditions play an important role during
the production of nanoparticles while using the fungi
cultures. When gold ions were incubated with the
Trichothecium sp. biomass under stationary condi-
tions led to the formation of extracellular nanopar-
ticles. While under shaking conditions, this was
resulted in the formation of intracellular gold nano-
particles. The possible reason for this could be the
enzymes and proteins responsible for the synthesis of
nanoparticles. These proteins were released into the
medium under stationary conditions and did not
release under shaking conditions (Ahmad et al.
2005). Bharde et al. (2006) reported the synthesis
of magnetic nanoparticles by using Fusarium oxy-
sporum and Verticillium sp. at room temperature.
Both fungi secreted proteins which were capable of
hydrolyzing iron precursors extracellularly to form
iron oxides predominantly in the magnetite (Fe3O4)
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phase. Also a nitrate-dependent reductase and a
shuttle quinone from several Fusarium oxysporum
strains were found to be involved in extracellular
production of silver nanoparticles or silver hydrosol.
However, it was not true with all Fusarium sp.
Fusarium moniliforme produces reductase enzyme
but could not form silver nanoparticles upon their
incubation with silver ions (Durán et al. 2005).
However, by controlling the amount of cofactor
NADH, synthesis of quite stable Au–Ag alloy
nanoparticles of various compositions have been
made possible. This approach can be further em-
ployed for producing various other composite nano-
particles (Senapati et al. 2005). Towards elucidating
the mechanism of synthesis of nanoparticles, a-
NADPH-dependent nitrate reductase and phytochel-
atin isolated from Fusarium oxysporum has been used
for in vitro silver nanoparticle production (Kumar
et al. 2007a).
Minimum time, miniaturization and non-hazard-
ous processes are key parameters for any kind of
technology acceptance. In this effort, Bhainsa and
D’Souza (2006) could get fairly monodispersed silver
nanoparticles within 10 min by using Aspergillus
fumigatus. This was the first report of such rapid
synthesis of nanoparticles using fungus. The produc-
tion was even faster compared to the physical and
chemical processes of nanoparticles synthesis. Hence,
this process could be suitable for developing a
biological process for mass scale production of
nanoparticles. Tetragonal barium titanate (BaTiO3)
nanoparticles of sub–10 nm dimensions produced by
Fusarium oxysporum under ambient conditions have
been observed an ecofriendly and economically
viable methods for the synthesis of complex oxide
nanomaterials of technological interest (Bansal et al.
2006). The presence of ferroelectric properties in
these nanoparticles would revolutionize the electronic
industries by making ultra small capacitors and
ultrahigh density nonvolatile ferromagnetic memo-
ries. Also, the synthesis of highly luminescent CdSe
quantum dots at room temperature, reported recently
by the fungus, Fusarium oxysporum when incubated
with a mixture of CdCl2 and SeCl4 would be of great
importance (Kumar et al. 2007b). Recently, Fusari-
um oxysporum fungus has also been used for the
production of silica and titania nanoparticles from
aqueous anionic complexes SiF6 2 and TiF6 2
respectively. Extra-cellular protein-mediated hydro-
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J Nanopart Res (2008) 10:507–517
lysis of the anionic complexes results in the facile
room temperature synthesis of crystalline titania
particles while calcination at 300 °C is required for
crystallization of silica (Bansal et al. 2005).
Use of plants
Quantum dots have huge application in nanobiotech-
nology and plants have been observed to be good
source for the synthesis of quantum dots. One of the
interesting investigations with Alfalfa documents the
feasibility for the synthesis of quantum dots by the
living plants. Alfalfa roots have capability for
absorbing Ag (0) from agar medium and transferring
them to shoot of the plant in the same oxidation state.
In the shoot, these Ag atoms arranged themselves to
form nanoparticles by joining themselves to form
larger arrangements. Transmission electron micro-
scope (TEM)/scanning electron microscope (SEM)
analysis showed that the accumulated Ag atoms
inside the plant tissue underwent nucleation and
resulted in the formation of nanoparticles (Gardea-
Torresdey et al. 2003). Furthermore, shape and size
variability can be induced in nanoparticles produced
by using plants by varying the pH of reaction
conditions. Armendariz et al. (2004) have reported
the pH-dependent binding trend of Au (III) ions to
Avena sativa (Oat) biomass and subsequent formation
of Au nanoparticles of variable sizes. Rod-shaped
nanoparticles were common at all pH values used
during the study. Larger nanoparticles (25–85 nm) in
small quantities were formed at pH 2, while smaller
nanoparticles (5–20 nm) in large quantities were
formed at pH 3 and 4. Mechanism for pH dependent
variability in shape and size of nanoparticles could be
such as Au (III) is present as an anion in aqueous
solutions and at low pH values, biomass might carry
more positive functional groups that allows the Au
(III) ions to get more closure to binding sites. Au (III)
as a soft metal binds to the biomass mainly through
the soft ligands. Also, positive amino and sulfhydryl
groups at low pH makes Au (III) available for the
binding and allow the reduction of Au (III) to Au (0).
Carboxylic groups, which are abundant in oat
biomass, are known to be protonated at low pH
might also be contribute to the binding of Au (III)
ions (Gardea-Torresdey et al. 2002). At low pH (2),
process of aggregation of Au nanoparticles to form
larger nanoparticles was favoured over the nucleation
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to form new nanoparticles. While at pH 3 and 4, more
functional groups could be available for gold binding,
thus a higher number of Au (III) complexes bind to
the biomass at the same time. This allowed the
subsequent formation of larger amounts of nanopar-
ticles with smaller diameters. Hence the size of
nanoparticles can be controlled by changing the pH
of the reaction conditions.
Similar to fungi, extracellular synthesis of nano-
particles is also possible using plants. Using Gera-
nium (Pelargonium graueolens) leaf extract silver
ions were reduced to silver nanoparticles. These
nanoparticles were highly stable and crystalline in
solution (Shankar et al. 2003). In addition to indi-
vidual pure metallic Ag and Au, bimetallic Ag/Au
nanopaticles (50–100 nm) using Azadirachta indica
leaf broth have also been synthesized extracellularly
(Shankar et al. 2004). Synthesis of bimetallic Au
core–Ag shell nanoparticles in solution was accom-
plished due to competitive reduction of Au3+ and Ag+
ions present simultaneously in the solution during the
exposure to Neem leaf extract. The rate of reduction
of metal ions by Neem leaf extract has been found to
be much faster compared to that of microorganisms
use. While, treatment of aqueous solutions of silver
nitrate and chloroauric acid with Neem leaf extract
has been observed for rapid formation of stable Ag
and Au nanoparticles at higher concentration. It was
believed that flavanone and terpenoid constituents of
leaf broth were the surface active molecules stabiliz-
ing the formation of nanoparticles, in contrast to high
molecular weight proteins in case of fungi.
With the use of Emblica Officinalis fruit extract as
reducing agent, the extracellular synthesis of highly
stable Ag and Au nanoparticles has been achieved
(Ankamwar et al. 2005a). Along with stability, the
control over the shape of nanoparticles production
using plants and parts their off is also possible. It has
been shown by the rapid synthesis of stable gold
nanotriangles at high concentration using tamarind
leaf extract as reducing agent (Ankamwar et al.
2005b). Similarly, using Aloe vera leaf extract as a
reducing agent the synthesis of gold nanotriangles as
well as silver nanoparticles was obtained in single
crystalline triangular form. For this, aqueous chlo-
roaurate ions were used with the Aloe vera plant
extracts. By varying the amount of extract in reaction
medium, percentage of gold nanotriangles to that of
spherical particles as well as the size of nanotriangles
513
(50–350 nm) have been modulated. The relative
amount of triangle and spherical nanoparticles led
to the significant control over the optical properties of
the nanoparticle solution (Chandran et al. 2006). The
key role in the formation of gold nanotriangles was
played by the slow reduction of aqueous gold ions
(HAuCl4) along with the shape-directing effects of
constituents (carbonyl compounds) of the plant
extract. On the other hand, aqueous silver ions
(AgNO3) when incubated with Aloe vera extract
produce only spherical Ag nanoparticles. The appear-
ance of brownish red colour and faint yellow colour
in the reactions indicate the formation of gold and
silver nanoparticles, respectively.
Adding to the list of plants which are showing
potential for nanoparticle production, Cinnamomum
camphora leaf extract has been identified very
recently for the production of gold as well silver
nanoparticles (Huang et al. 2007). The marked dif-
ference of shape control between gold and silver
nanoparticles was attributed to the comparative
advantage of protective biomolecules and reductive
biomolecules. The polyol components and the water-
soluble heterocyclic components were mainly found
to be responsible for the reduction of silver ions or
chloroaurate ions and the stabilization of the nano-
particles, respectively.
Application of nanoparticles
Development of nano-devices using biological mate-
rials and their use in wide array of applications on
living organisms has recently attracted the attention
of biologists towards nanobiotechnology. Below
mentioned are few applications of this technology
which would help in understanding the use of diverse
living organisms in nanodevices production and also
the use of these nanoproducts in various applications.
Nanotechnology could be useful to make nano-
scale confining environments channels or post arrays
for long polymers such as DNA (Austin et al. 2002).
Gene expression could be enhanced by increasing the
delivery of intact DNA to cell nuclei by producing
hybrid polymer–protein conjugate nanoparticles. It
could also be used in building automated nano-chip
sensors, therapeutic and diagnostic devices, where
DNA itself would be a target, or where DNA might
be used as a ‘conveyor-belt’ for attached molecules.
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Various synthetic polymers such as molecular motor
with ‘smart’ polymers could be prepared by using
nanotechnological methods to regulate biological
processes (Pennadam et al. 2004).
The sensitivity and performance of biosensors
have been improved by using nanomaterials. Many
new signal transduction technologies have been
introduced in biosensors, bioprobes and other bio-
systems using nanomaterials produced through living
organisms (Jianrong et al. 2004). The gold nanotri-
angles with unique and highly anisotropic planer
shapes might find application in photonics, optoelec-
tronics, and optical sensing. Further, the effect of
different organic solvent vapors like methanol, ben-
zene and acetone on the conductivity of tamarind leaf
extract reduced gold nanotriangles suggests the
application of gold nanotriangles to future chemical
sensors (Ankamwar et al. 2005b).
Bioremediation of radioactive wastes, resulted
from nuclear power plants and nuclear weapon
production such as Uranium (a long- lived radionu-
clide hazardous for both flora and fauna) have been
achieved by using nanoparticles. Cells and S-layer
proteins of Bacillus sphaericus JG-A12 have special
capabilities for the clean-up of uranium contaminated
waste waters. The effluent derived from the several
processes can be bioremediated by treating with C.
violaceum (Durán et al. 2007).
Recently, nanoparticles have found applications in
antibacterial effects. It has been shown that extracel-
lularly produced silver or gold nanoparticles using
Fusarium oxysporum, can be incorporated in several
kinds of materials such as cloths. These cloths with
silver nanoparticles are sterile and can be useful in
hospitals to prevent or to minimize infection with
pathogenic bacteria such as Staphylococcus aureus
(Durán et al. 2007).
Gold and silver nanoparticles produced either
intra- or extra-cellularly by using living organisms
could be of great value. Silver nanoparticles have
large number of applications such as in non-linear
optics, spectrally selective coating for solar energy
absorption, biolabelling (Klaus 2001), intercalation
materials for electrical batteries, as optical receptors,
catalyst in chemical reactions and as antibacterial
capacities (Durán et al. 2005). Magnetite (Fe3O4)/
greigite (Fe3S4) and siliceous material produced from
magnetotactic bacteria and diatoms, respectively, are
used in optical coatings for solar energy applications
123
J Nanopart Res (2008) 10:507–517
and as ion insertion materials for electrical battery
applications (Joerger et al. 1999).
The filamentous fungus, A. fumigatus has been
known for rapid extracellular synthesis of fairly
monodispersed silver nanoparticles ranging from 5 to
25 nm. Hence, this system could be suitable for
developing a biological process for mass scale
production. Furthermore, the extracellular synthesis
would make the process simpler and easier for
downstream processing. In future, it might be
important to understand the biochemical and molec-
ular mechanism of the synthesis of the nanoparticles
by the cell filtrate in order to achieve better control
over size and polydispersity of the nanoparticles
(Bhainsa and D’Souza 2006). The fungus, Aspergil-
lus flavus when challenged with silver nitrate solution
also accumulated reasonably monodisperse silver
nanoparticles on the surface of its cell wall (Vig-
neshwaran et al. 2007). Hence, the use of fungus for
silver nanoparticles synthesis offers the benefits of
ecofriendliness and amenability for large-scale pro-
duction. A variety of bacteria, yeast and fungi have
the ability to produce various kind of nanoparticles.
In addition, by manipulations of key parameters,
which control growth and other cellular activities,
controlled size and shape of the nanoparticles can be
achieved (Gericke and Pinches 2006a, b).
Biominerals have been formulated by using sev-
eral bacteria such as Pseudomonas aeruginosa (Mul-
len et al. 1989), E. coli (Bayer and Bayer 1991) and
Citrobacter sp. Metal sulphide microcrystallites were
formulated by using S. pombe, which can function as
quantum semiconductor crystallite. These crystallites
also have properties like optical absorption, photo-
synthetic and electron transfer. Furthermore, the
regenerative capability of biological systems coupled
with the discovery that fungi such as Fusarium
oxysporum are capable of hydrolyzing metal com-
plexes that they never encounter during their growth
cycle shows enormous promise for development,
particularly the large-scale synthesis of metal oxide
semiconductor materials (Bansal et al. 2005). While,
magnetosome particles isolated from magnetotactic
bacteria have been used as carrier for immobilization
of bioactive substances such as enzymes, antibodies,
DNA and RNA (Matsunaga 1991).
Contarary to a number of chemical methods used
in the synthesis of quantum dots that are often energy
intensive, employ toxic chemicals, and require higher
J Nanopart Res (2008) 10:507–517
temperatures, the synthesis by biological systems has
been characterized by processes that occur at ambient
temperature and pressure. Synthesis of highly lumi-
nescent CdSe quantum dots at room temperature has
been reported by the fungus, Fusarium oxysporum
when incubated with a mixture of CdCl2 and SeCl4
(Kumar et al. 2007b).
Nitrate reductase from Fusarium oxysporum has
been shown to catalyze the production of stable silver
nanoparticles under in vitro condition. This study has
opened the way for designing a rational enzymatic
strategy for the synthesis of nanomaterials of differ-
ent chemical composition, shapes and sizes as well as
their separation (Kumar et al. 2007a). Silver nano-
particles and triangular or spherical shaped gold
nanoparticles have been produced by using sundried
biomass of Cinnamomum camphora leaf with aque-
ous silver or gold precursors at ambient temperature.
The marked difference of shape control between gold
and silver nanoparticles was attributed to the com-
parative advantage of protective biomolecules and
reductive biomolecules. The polyol components and
the water-soluble heterocyclic components were
mainly responsible for the reduction of silver ions
or chloroaurate ions and the stabilization of the
nanoparticles, respectively. Therefore, such chemical
components could be useful for simple synthesis of
nanoparticles (Huang et al. 2007).
Small interfering RNA (siRNA) delivery can be
monitored by a novel method based on nanodevice that
combines unmodified siRNA with semiconductor
quantum dots (QDs) as multicolour biological probes.
Co-transfection of siRNA with QDs using standard
transfection techniques has led to the formation of
photostable fluorescent nanoparticles that helps in
tracking the delivery of nucleic acid, the degree of
transfection in cells and also in purifying homoge-
neously silenced subpopulations (Chen et al. 2005).
Use of nanodevices in siRNA delivery to the system
have also improved the silencing efficiency. The
delivery of oligocarbamate conjugate siRNA (siRNA-
TAT47–57 peptide) into cells has resulted in efficient
RNAi activity. Further nanoparticles/nanodevice also
helps in loalization of siRNA activity and in this
particular example, siRNA activity was localized in
the perinuclear space of the cells (Chiu et al. 2004).
The controlled release of siRNA has also been
achieved by using nanodevices. Organic-inorganic
hybrid nanoparticles entrapping oligodeoxynucleotide
515
(ODN) or siRNA have been easily taken up by the cells
and therein release the incorporated ODN and siRNA
in highly controlled manner (Kakizawa et al. 2004).
Conclusion and future perspectives
Living organisms have huge potential for the pro-
duction of nanoparticles/nanodevices of wide appli-
cations. By using the organisms from simple bacteria
to highly complex eukaryotes in the reaction mixture,
the production of nanoparticles/nanodevices with
desired shape and size can be obtained. Though
nano-biotechnology is at its infancy but various
examples through which this technology and their use
have been explained in this article would attract the
attention of peoples towards its applications. Among
these studies, a few have shown that different kind of
reductases of these organisms might be involving in
the mechanism of nanoparticles production and
attribute them various shape and size. However, the
elucidation of exact mechanism of nanoparticles
production using living organisms needs much more
experimentations.
Acknowledgements Authors are thankful to Dr. P.S. Ahuja,
Director, IHBT for his valuable suggestions during writing of
this article. We would like to acknowledge Council for
Scientific and Industrial Research (CSIR), Govt. of India,
New Delhi for providing continuous financial support for our
research work. The IHBT communication number of this
article is 755.
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