Colloids and Surfaces B: Biointerfaces 103 (2013) 283287

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Review
A review on biosynthesis of nanoparticles by marine organisms
N. Asmathunisha, K. Kathiresan

Centre of Advanced Study in Marine Biology, Faculty of Marine Sciences, Annamalai University, Parangipettai 608502, India
a r t i c l e i n f o
Article history:
Received 6 August 2012
Accepted 17 October 2012
Available online 30 October 2012
Keywords:
Marine
Nanoparticles
Mangroves
Salt marshes
Seaweeds
Algae
a b s t r a c t
Marine organisms produce remarkable nanofabricated structures in cell wall, shells, pearls and sh bones.
Marine microorganisms such as bacteria (E. coli, Pseudomonas sp.), cyanobacteria (Spirulina platensis,
Oscillatoria willei, Phormidium tenue), yeasts (Pichia capsulata, Rhodospiridium diobovatum), fungi (Thraus-
tochytrium sp., Penicillium fellutanum, Aspergillus niger), and algae (Navicula atomus, Diadesmis gallica,
Stauroneis sp. Sargassum wightii, Fucus vesiculosus) are reported to synthesize inorganic nanoparticles
either inside or outside cells. Mangroves (Rhizophora mucronata, Xylocarpus mekongensis), salt marshes
(Sesuvium portulacastrum and Suaeda sp.) and sand dune (Citrullus colocynthis) are also capable of syn-
thesizing the nanoparticles, in addition to marine animals such as nsh and sponges. Biosynthesis
of nanoparticles may be triggered by several compounds such as carbonyl groups, terpenoids, pheno-
lics, avonones, amines, amides, proteins, pigments, alkaloids and other reducing agents present in
the biological extracts. Marine bio-nanotechnology has a great promise in nanomedicines, food stuff,
pharmaceuticals and fabric industries for the future.
2012 Elsevier B.V. All rights reserved.
Contents
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 283
2. Types of nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 283
3. Biological synthesis of nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 284
4. Biosynthesis of nanoparticles by marine microorganisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 284
5. Biosynthesis of nanoparticles by marine spermatophytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 285
6. Biosynthesis of nanoparticles by marine algae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 285
7. Biosynthesis of nanoparticles by marine animals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 285
8. Applications of nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 285
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 286
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 286
1. Introduction
Marine bio-nanotechnology is an exciting and upcoming area of
research. The biologically diverse marine environment has a great
promise for nanoscience and nanotechnology. Marine organisms
produce remarkable nanoparticles of 1100nmsize which consti-
tute nanofabric structures such as seashells, pearls and sh bones.
Diatoms and sponges are constructed with nanostructured cover of
silica and coral reefs with calcium, arranged in remarkable archi-
tectures [1]. Dolphins and whales have rough skin surface due to
presence of nanoridges. These ridges enclose a pore size of 0.2m
2
which is below the size of marine fouling organisms and hence,

Corresponding author. Tel.: +91 4144 243223x210; mobile: +91 9442068003.

E-mail address: kathirsum@rediffmail.com (K. Kathiresan).
there is no attachment of biofoulers [2]. In spite of great potential,
the marine bio-nanotechnology has limited research work. Most of
the studies on biosynthesis of nanoparticles have been restricted
to terrestrial organisms. This review was synthesized on available
information on the marine biosynthesis of nanoparticles and their
mechanism and biological activities to bring out the prospects of
marine nanobiotechnology.
2. Types of nanoparticles
Nanoparticles are classied into major types viz. organic
and inorganic nanoparticles. Carbon nanoparticles are called
the organic nanoparticles. Magnetic nanoparticles, noble metal
nanoparticles (platinum, gold and silver) and semiconductor
nanoparticles (titanium dioxide and zinc oxide) are grouped as
inorganic nanoparticles. Inorganic nanoparticles are increasingly
0927-7765/$ see front matter 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.colsurfb.2012.10.030
284 N. Asmathunisha, K. Kathiresan / Colloids and Surfaces B: Biointerfaces 103 (2013) 283287
Table 1
Outline of work on biosynthesis of nanoparticles using marine organisms.
Organism Species Name of the
species
Types of
nanoparticles
Size (nm) Biological activity Author and year
Marine microbes Cyanobacteria Spirulina platensis Silver 716

Govindaraju et al.
(2008) Gold 610
Biometallic 1725
Oscillatoria willei Silver 100200 Mubarak et al. (2011)
Phormidiumtenue Cadmium 5 Mubarak et al. (2012)
Bacteria E. coli Silver 520 Antimicrobial Kathiresan et al. (2010)
Pseudomonas sp. Silver 20100 Muthukannan and
Karuppiah (2011)
Yeast Pichia capsulata Silver 50100 Manivannan et al. (2010)
Rhodosporidium
diobovatum
Lead 25 Sesadhri et al. (2011)
Fungi Penicillium
fellutanum
Silver 520 Kathiresan et al. (2009)
Thraustochytrium
sp.
Silver Gomathi (2009)
Aspergillus niger Silver 535 Antimicrobial Kathiresan et al. (2010)
Diatoms Navicula atomus,
Diadesmis gallica
Gold 9 Adamet al. (2011)
Goldsilica 22
Stauroneis sp. Silicongermanium Mubarak et al. (2011)
Marine algae Seaweed Sargassumwightii Gold 812 Singaravelu et al. (2007)
Sargassumwightii Silver Antibacterial Govindaraju et al. (2009)
Turbinaria conoides Silver Fabric
strengthening
Mercy Sheeba and
Thambidurai (2009)
Gelidiella acerosa Silver 22 Antifungal Vivek et al. (2012)
Ulva fasciata Silver 2841 Antibacterial Rajesh et al. (2012)
Brown alga Fucus vesiculosus Gold Biosorption Mata et al. (2008)
Marine alga Cladosiphon
okamuranus
Kjellamaniella
crassifolia
Gold 8.5410.74 Suwicha et al. (2010)
Marine spermatophytes Mangroves Xylocarpus
mekongensis
Silver 520 Antimicrobial Asmathunisha et al. (2010)
Rhizophora
mucronata
Silver 6095 Larvicidal Gnanadesigan et al. (2011)
Salt marshes Sesuvium
portulacastrum
Silver 5090 Antimicrobial Asmathunisha (2010)
Sand dune Citrullus colosynthis Silver 85100 Anticancer Satyavani et al. (2011)
Coastal plant Prosopis chilensis Silver 525 Antibacterial to
control vibriosis in
Penaeus monodon
Kathiresan et al. (2012)
Marine animals
Sponges Acanthella elongata Gold 720 Inbakandan et al. (2010)
Fin sh Cod liver oil Silver 510 Khanna and Nair (2009)
usedindrugdeliverydue totheir distinctive features suchas ease of
use, goodfunctionality, biocompatibility, abilitytotargetedspecic
cell and controlled release of drugs [3].
3. Biological synthesis of nanoparticles
Use of chemical and physical method in the synthesis of
nanoparticles is very expensive and cumbersome. The chemical
and physical methods of nanoparticle synthesis lead to the pres-
ence of some toxic chemicals absorbed on the surface that may
have adverse effects in applications, so there is a growing need to
develop environmentally benign nanoparticles. Researchers have
usedbiological extracts for thesynthesis of nanoparticles, byadopt-
ing simple protocols, involving in the process of reduction of metal
ions by using biological extracts as a source of reductants either
extracellularly or intracellularly.
Synthesis of nanoparticles may be triggered by several com-
pounds such as carbonyl groups, terpenoids, phenolics, avonones,
amines, amides, proteins, pigments, alkaloids and other reducing
agents present in the plant extracts and microbial cells [48]. The
exact mechanismof nanoparticles synthesis by biological extracts
is yet to be understood.
4. Biosynthesis of nanoparticles by marine microorganisms
Microorganisms suchas bacteria, cyanobacteria, actinomycetes,
yeast, fungi, and algae are known to synthesize inorganic nanopar-
ticles such as gold, silver, calcium, silicon, iron, gypsum and lead,
in nature either inside or outside cells. At present, microbial meth-
ods in the synthesis of nanomaterials of varying compositions are
extremely limited and conned to metals, some metal sulde, and
very low oxides. All these are restricted to the microorganisms of
terrestrial origin. Marine microbes have potential ability to synthe-
sis nanoparticle for the reasonthat the marine microbes exist inthe
sea bottom, over millions of years in the past for reducing the vast
amount of inorganic elements deep in the sea. It is important to
study the marine microbes for biosynthesis of nanoparticles and
to elucidate biochemical pathways that lead to metal ion reduc-
tion by the different classes of microbes to develop nanoparticles.
The biosynthesis of nanoparticles with the use of microorgan-
isms depends on culture conditions and hence standardizing these
conditions for high synthesis of nanoparticles is necessary. Many
marine microorganisms are known to produce nanostructured
mineral crystals and metallic nanoparticles with properties similar
to chemically synthesized materials, while exercising strict control
over size, shape and composition of the particles (Table 1).
N. Asmathunisha, K. Kathiresan / Colloids and Surfaces B: Biointerfaces 103 (2013) 283287 285
The marine fungus Penicillium fellutanum isolated from coastal
mangrove sediment extracellularly produces silver nanoparti-
cles, when it is exposed to AgNO
3
[9]. In spite of the fact that
Thraustochytrids, obligate marine fungi are abundant with poly
unsaturated fatty acids [1012]. Their application in nanoparticles
is not known, until. Gomathi [13] demonstrates the extracellular
biosynthesis of silver nanoparticles using Thraustochytriumsp., and
the formation of lipid/silver nanoparticle composites.
The strains of Escherichia coli AUCAS 112 and Aspergillus niger
AUCAS237isolatedfrommangrovesediments arecapableof reduc-
ing the silver ions in faster rate. The antimicrobial activity of
nanoparticles produced fromE. coli is more pronounced than that
of A. niger and is also enhanced with the addition of polyvinyl
alcohol as a stabilizing agent. The synthesized silver nanoparti-
cles are monodispersed and spherical in nature [14]. Whereas, a
novel strain of Psuedomonas sp. 591786 has been shown to pro-
duce the intracellular silver nanoparticles which are polydispersed
with different size groups ranging from20 to 100nm[15].
The culture ltrate of mangrove-derived yeast Pichia capsulata
exhibits the most efcient productionof silver nanoparticles within
minutes [16]. The protein present in the culture ltrate of the
yeast species is responsible for the synthesis of silver nanopar-
ticles [16]. But sulphur rich peptide present in the marine yeast
Rhodosporidiumdiobovatumacts as a capping agent for synthesis of
lead sulphide nanoparticles [17].
Govindaraju et al. [18] have studied the production of silver,
gold and bimetallic nanoparticles production using single-cell pro-
tein, Spirulina platensis. Fourier transform infrared spectroscopic
measurement reveals the fact that the protein is the possible
biomolecule responsible for the reduction and capping of the
biosynthesized nanoparticles. The marine cyanobacterium, Oscil-
latoria willei NTDM01 is known to secrete a protein which is
responsible for reduction of silver ions and stabilization of silver
nanoparticles [19]. Very recently Mubarak et al. (2012) [20] have
reported the synthesis and characterization of cadmium sulphide
(CdS) nanoparticles using C-phycoerythrin (C-PE) extracted from
the marine cyanobacterium, Phormidium tenue NTDM05. The size
of the CdS nanoparticles is found to be about 5nm. Essentially, it is
found that the pigment stabilized the CdS nanoparticles. The pig-
ments labeled CdS nanoparticles can also be applied as a biolabel.
5. Biosynthesis of nanoparticles by marine spermatophytes
Recently, coastal plants are known to synthesis nanoparticles.
While screening 26 plants of coastal origin for silver nanoparticle
synthesis, Asmathunisha[21] has observedthehighest synthesis by
the mangrove Xylocarpus mekongensis followed by the salt marsh
Suaeda maritima. In addition to intact plant extracts, callus is also
known to better produce nanoparticles. This has been proved with
synthesis of antimicrobial silver nanoparticles by leaf callus better
than leaf extracts of the salt marsh plant, Sesuviumportulacastrum
[22]. Flavonones and terpenoids are likely responsible for the sta-
bilization of the silver nanoparticles. The silver nanoparticles do
inhibit clinical strains of bacteria and fungi. Antibacterial activ-
ity of the nanoparticles is more distinct than antifungal activity
[22]. The silver nanoparticles have also been proved to have anti-
cancer property in the sand dune plant, Citrullus colocynthis when
thenanoparticles synthesizedbycallus extract of theplant has been
tested on the cell line of human epidermoid larynx carcinoma (HEp
-2) by using MTT assay, caspase-3 assays, lactate dehydrogenase
leakage assay and DNA fragmentation assay [23]. In addition to
the antimicrobial and anticancer properties, silver nanoparticles
also exhibit mosquito larvicidal activity. The nanoparticles synthe-
sizedbythemangroveleaf extract of Rhizophoramucronataexhibits
larvicidal activity against Aedes aegypti and Culex quinquefasciatus
withthe LC
50
values of 0.585and0.891mg/L respectively [24]. Very
recently, the application of silver nanoparticles is reported in con-
trolling shrimp diseases such as vibriosis. The silver nanoparticles
produced by the coastal Prosopis chilensis inhibit vibrio pathogens
viz., Vibrio cholerae, V. harveyi, and V. parahaemolyticus and this
antibacterial effect of nanoparticles is better thanthat of leaf extract
as proved by disc diffusion assay. The nanoparticles are then tested
in the shrimp challenged with the four species of vibrio pathogens
for 30days. Theshrimps fedwithsilver nanoparticles exhibit higher
survival, associated with immunomodulation in terms of higher
haemocyte counts, phenoloxidase and antibacterial activities of
haemolymph of the tiger shrimp, Penaeus monodon [25].
6. Biosynthesis of nanoparticles by marine algae
There is a verylittle literature supportingthe use of marine algae
in nanoparticle synthesis. The brown seaweed Sargassumwightii is
reportedly capable of synthesizing gold nanoparticles with a size
ranging between 8 and 12nm. An important potential benet of
the synthesis is that the nanoparticles are quite stable [26]. Another
brown seaweed Fucus vesiculosus is reported to have an ability of
gold biosorption and bioreduction, as an environmental friendly
process that can be used for recovering gold fromdilute hydromet-
allurgical solutions and leachates of electronic scraps, and for the
synthesis of gold nanoparticles of different size and shape [27].
Similarly, the extracellular synthesis of silver nanoparticles by the
brownseaweedSargassumwightii andtheir antibacterial effects are
registered [28]. In addition to antibacterial activity, the nanoparti-
cles synthesized by seaweed extracts do have stabilizing effect on
cotton fabrics [29]. Fucoidan is an algal polysaccharide, reported
to stabilize gold particles and this green synthesis using natural
fucoidans will provide an alternative to chemical method [30]. The
red seaweed Gelidiella acerosa is reported to have the potential of
synthesizing antifungal silver nanoparticles [31]. Recently, Rajesh
et al. have reported the synthesis of silver nanoparticles using Ulva
fasciata extract as a reducing agent and this nanoparticles inhibited
the growth of Xanthomonas campestris pv. malvacearum [32]. In
additiontoseaweeds, microalgaesuchas diatoms (Naviculaatomus,
Diadesmis gallica) have the ability to synthesize gold nanopar-
ticles, gold, and silicagold bionanocomposites [33]. The diatom
Stauroneis sp. was used for the preparation of silicongermanium
nanocomposite and this method of nanocomposite preparationhas
great importance for possible future applications due to its acces-
sibility, simplicity and effectiveness [34].
7. Biosynthesis of nanoparticles by marine animals
Fishoil is of neutraceutical valueandthepresenceof permissible
limit of silver nanoparticles inthe oil might enhance its efcacyan
idea that may open many avenues in the eld of nanobiotech-
nology. The use of cod liver sh oil is shown to produce silver
nanoparticles, as reducing agent as well as surfactant. Presence of
carboxylateions andaminegroups intheshoil triggers insitugen-
eration of organically capped silver nanoparticles [35]. The marine
sponge, Acanthella elongata is shown to produce gold nanoparticles
and this process is attributed to water-soluble organics present in
the sponge extract [36].
8. Applications of nanoparticles
Nanoparticles have a greater surface area per weight than larger
particles and this property makes them to be more reactive to
certain other molecules and they are used or being evaluated for
use in many elds [9]. Quantum dots are the crystalline nanopar-
ticles used to identify the location of cancer cells in the body.
286 N. Asmathunisha, K. Kathiresan / Colloids and Surfaces B: Biointerfaces 103 (2013) 283287
Gold nanoparticles allowheat frominfrared lasers to detect cancer
tumors. The iron oxide nanoparticles are used in better diagnosis
of tumors by magnetic resonance imagining (MRI) scan. Once the
nanoparticles are attached to the tumor, their magnetic property
enhances the images of the scan [37]. In addition to diagnosis, the
nanoparticles are usedindrug delivery andremoval of newtumors.
Magnetic nanoparticles that attach to cancer cells in the blood
streammay allowthe cancer cells to be removed before they estab-
lish new tumors [38]. The nanoparticles coated with proteins can
be attachedto damagedportions of arteries. This canallowdelivery
of drugs directly to the damaged portion of arteries to ght against
cardiovascular disease. Combining gold nanoparticles with organic
molecules to create a transistor is known as an OMFET (nanopar-
ticle organic memory eld-effect transistor) [39,40]. Intravenous
injection of gold nanoparticles (2nm in diameter) can enhance
radiotherapy (X rays) and results in eradication of subcutaneous
mammary tumors in mice and it is proved to increase survival of
the test animal to 86%as against 20%withX-rays alone. Apparently,
gold nanoparticles are non-toxic to mice and are cleared fromthe
body through the kidneys [41].
Chitosan is an important component of exoskeleton of marine
crustaceans and fungi. The chitosan nanoparticles exhibit potential
antibacterial activity [42]. They do have optical sensing properties
and can also easily bind with other molecules. This has potential to
develop nanotechnology devices in detecting avain u, breast can-
cer, and also toxic substances in air, water and soil. The chitosan
nanomaterial incorporated with drug can be prepared in micro-
capsules. These will open with specic biochemical activity related
to tumor. Since the chitosan nanoparticles can be easily detected
as the uorescence when exposed near infra red light. Using the
technique, even tiny breast cancer of 12mmcan be detected [2].
These are possibilities for production of lipid nanoparticles
with the help of lipid-rich marine organisms such as algae, fungi
and bacteria [13]. Lipid nanoparticles can be synthesized from
the organisms through heating to liquefy fatty acids; incorporat-
ing active agents of pharmacological and cosmetics importance;
adding a hot surfactant; and stirring or homogenized under high
pressure by ultra sound. These can be used in the production of
food stuffs, cosmetics and medicines [2].
Silica is an essential component of cell wall of diatoms, dinoag-
ellates and many invertebrates such as sponges, but rarely used for
nanoparticle synthesis. Silicate nanoparticles are used to provide
a barrier to gas (for example oxygen), or moisture in a plastic lm
used for packaging. This can reduce the possibly of food spoiling or
drying. Silicon nanoparticles coating anodes of lithium-ion batter-
ies increase battery power and reduce recharge time [43]. Silicon
dioxide crystalline nanoparticles lling gaps between carbon bers
strengthen their stability in tennis racquets [44,45]. Besides sil-
ica, silver nanoparticles have possible application in fabrics as they
kill bacteria and thus make clothing odor-resistant [46]. Zinc oxide
nanoparticles dispersed in industrial coatings protect wood, plastic
andtextiles fromexposure toUVrays [47,48]. The nanoparticles are
also used to remove pollution. For example, iron nanoparticles can
clean up carbon tetrachloride pollution in ground water [49]. The
nanoparticle coating composed of titaniumdioxide (TiO
2
) and sil-
ver nanoparticles is reportedly preventing biofouling of ship hulls.
This biofouling of ship hulls is a very serious problem in fuel con-
sumption, biocorrosion and ultimately heavy economic loss to the
shipping industry throughout the world [50].
The marine extracts have not received due attention for biosyn-
thesis of nanoparticles, even though the marine organisms are
rich sources of reducing agents, precursor molecules such as silica,
calcium, chitosan, and also stabilizing agents such as polysaccha-
rides, lipids and peptides. In the present context of increasingly
recognized utility of nanoparticles in cancer treatments, toxicity
of nanoparticles synthesized by various marine extracts on cancer
cells and normal cells has to be largely studied and their mech-
anism(s) of actions have to be elucidated in terms of antioxidant
potential and induction of apoptosis.
Acknowledgements
The authors are thankful to authorities of Annamalai University
and to UGC, NewDelhi for nancial assistance (MANF).
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