BIOLOGICAL SYNTHESIS

Biosynthesis of metal nanoparticles is an environmentally

friendly method (green chemistry) without use of harsh,
toxic and expensive chemicals.
Nanoparticles have been produced physically and
chemically for a long time, but recent developments show
the critical role of microorganisms and biological systems
in production of metal nanoparticles.
In recent years, the development of efficient green
chemistry methods for synthesis of metal nanoparticles has
become a major focus of researchers.
They have investigated in order to find an eco-friendly
technique
for
production
of
well-characterized
nanoparticles.
Biosynthesis of nanoparticles gained lots of interest due to
the use of mild experimental conditions such as
temperature, pH and pressure.
One of the most considered methods is production of metal
nanoparticles using organisms.
The ability of organisms in production of metal
nanoparticles has opened a new exciting approach toward
the development of these natural nano-factories.
The organisms used in nanoparticle synthesis vary from
simple prokaryotic bacterial cells to complex eukaryotes.
The important aspects which might be considered in the
process of producing highly stable and well-characterized
nanoparticles are as follows:

1. Selection of the best organisms:

In order to choose the best candidates for metal
nanoparticle production, researchers have focused on
the important intrinsic properties of the organisms
such as enzyme activities and biochemical pathways.
For example, plants which have great potential in
heavy metal accumulation and detoxification are the
best candidates for nanoparticle synthesis.
2. Optimal conditions for cell growth and enzyme activity:
Optimization of the growth conditions is very
important. The nutrients, inoculum size, light,
temperature, pH, mixing speed, and buffer strength
should be optimized.
The presence of the substrates or related compounds
in subtoxic levels from the beginning of the growth
would increase the activity of the enzymes.
3. Optimal reaction conditions:
In order to use the organisms for synthesis of metal
nanoparticle in industrial scale, the yield and the
production rate are important issues to be considered.
Therefore, we need to optimize the bioreduction
conditions in the reaction mixture.
The substrate concentration, the biocatalyst
concentration, the electron donor and its
concentration, pH, exposure time, temperature, buffer
strength, mixing speed, and light need to be controlled
and optimized.

 Moreover,
researchers
have
used
some
complementary factors such as visible light or
microwave irradiation, and boiling which could affect
the size, morphology, and rate of reaction.
Mechanism
The most important mechanism of metal nanoparticles
biosynthesis by microorganisms is bioreduction.
In
microbial
bioreduction
processes,
proteins,
carbohydrates and biomembranes are involved.
Nanoparticles are formed on cell wall surfaces, and the first
step in bioreduction is the trapping of the metal ions on this
surface.
This probably occurs due to the electrostatic interaction
between the metal ions and positively charged groups in
enzymes present at the cell wall.
This may be followed by enzymatic reduction of the metal
ions, leading to their aggregation and the formation of
nanoparticles.
The microbial cell reduces metal ions by use of specific
reducing enzymes like NADH-dependent reductase or
nitrate-dependent reductase.
Reductases involved in NP synthesis through microbes
Oxidoreductases- Yeast
NADH-dependent reductase- Fusarium oxysporum, Aspergillus
flavus

Nitrate/nitrite

reductase-

Fusarium oxysporum, Klebsiella

pneumonia, E. coli and Enterobacter cloacae
Sulfate and sulfite reductase- Rhodobacter sphaeroides,
Plectonema boryanum (Cyanobacteria)

BACTERIA
One approach that shows immense potential is based on the
biosynthesis of NPs using bacteria.
A wide number of bacterial species have been used in green
nanotechnology to research alternative methods for the
synthesis of NPs.
Researchers have started to use biomass or cell extracts of
bacteria for synthesizing NPs.
Bacteria are considered as a potential biofactory for the
synthesis of NPs like gold, silver, platinum, palladium,
titanium, titanium dioxide, magnetite, cadmium sulphide,
and so forth.
Some well-known examples of bacteria synthesizing
inorganic materials include magnetotactic bacteria (which
orient along the magnetic field lines of Earth's magnetic
field using
structures called magnetosomes; Ex:
Magnetospirillum magneticum) and S layer bacteria.
Most metal ions are toxic for bacteria, and, therefore, the
bioreduction of ions or the formation of water insoluble
complexes is a defense mechanism developed by the
bacteria to overcome such toxicity.

Mechanism:
Shewanella alga is a gram-negative bacillus, which is widely
distributed in the environment, and its natural habitats are
water and soil.
Used to produce gold nanoparticles.

 The microbial reduction of gold ions was dependent on the

presence of a specific electron donor, the molecular H2.
It was concluded that the S. alga hydrogenase catalyzes the
activation of molecular H2 using the molecule as the
electron donor according to the following reaction:

The S. algae cells are likely to transfer electrons to AuCl4

ions, reducing them to gold metals.

Future prospects
Major drawbacks associated with the biosynthesis of NPs
using bacteria are tedious purification steps and poor
understanding of the mechanisms.
The important challenges frequently encountered in the
biosynthesis of NPs are to control the shape and size of the
particles and to achieve the monodispersity in solution
phase.
It seems that several important technical challenges must be
overcome before this green bio-based method will be a
successful and competitive alternative for industrial
synthesis of NPs.
An important challenge is scaling up for production level
processing.
Furthermore, little is known about the mechanistic aspects,
and information in this regard is necessary for economic
and rational development of nanoparticle biosynthesis.

FUNGI
Compared with bacteria, fungi have been known to secrete
much higher amounts of bioactive substances, which made
fungi more suitable for large-scale production.
The extracellular biosynthesis using fungi could also make
downstream processing much easier than bacteria.
An interesting example of the biosynthesis using fungi was
that the cell-associated biosynthesis of silver using
Fusarium oxysporum was demonstrated by Ahmad et al.,
and the particles were overall quasi-spherical with size
range between 5 and 15 nm.
There also have been several reports on the biosynthesis of
AgNPs using fungi, including Fusarium acuminatum and
Penicillium fellutanum.

Figure: Colour change during nanoparticle formation (The crude cell filtrate of
Aspergillus terreus mixed without AgNO3 (A) and with AgNO3 (B) after 24 h)

PLANTS
Among these organisms plants seem to be the best
candidates and they are suitable for large-scale
biosynthesis of nanoparticles.
Nanoparticles produced by plants are more stable and the
rate of synthesis is faster than in the case of
microorganisms.
Moreover, the nanoparticles are more various in shape and
size in comparison with those produced by other
organisms.
The advantages of using plant and plant-derived materials
for biosynthesis of metal nanoparticles have interested
researchers to investigate mechanisms of metal ions uptake
and bioreduction by plants, and to understand the possible
mechanism of metal nanoparticle formation in plants.
Gardea-Torresdey et al. (2002, 2003) reported for the first
time the formation of gold and silver nanoparticles inside
living plants.
They demonstrated the synthesis of gold and silver
nanoparticles within live Medicago sativa (alfalfa) plants by
gold and silver ion uptake, respectively, from solid media.
Synthesis of metal nanoparticles using plant extracts is
very cost effective, and therefore can be used as an
economic and valuable alternative for the large-scale
production of metal nanoparticles.
Current research in biosynthesis of nanometals using plant
extracts has opened a new era in fast and nontoxic methods
for production of nanoparticles.

 The bioreduction of metal nanoparticles by combinations

of biomolecules found in plant extracts (e.g. enzymes,
proteins, amino acids, vitamins, polysaccharides, and
organic acids such as citrates) is environmentally benign,
yet chemically complex but poorly understood.
Many biomolecules in plants such as proteins/enzymes,
aminoacids,
polysaccharides,
alkaloids,
alcoholic
compounds, and vitamins could be involved in
bioreduction, formation and stabilization of metal
nanoparticles.
Reduction potential of ions and reducing capacity of plants
which depend on the presence of polyphenols, enzymes,
and other chelating agents present in plants have critical
effects on the amounts of nanoparticle production.

Conclusion
Plants (especially plant extracts) are able to reduce metal
ions faster than fungi or bacteria.
Furthermore, in order to use an easy and safe green
method in scale-up and industrial production of welldispersed metal nanoparticles, plant extracts are certainly
better than plant biomass or living plants.