Fabrication of Metal and Metal Oxide Nanoparticles by Algae and their Toxic Effects
© The Author(s). 2016
Received: 19 May 2016
Accepted: 12 August 2016
Published: 17 August 2016
Of all the aquatic organisms, algae are a good source of biomolecules. Since algae contain pigments, proteins, carbohydrates, fats, nucleic acids and secondary metabolites such as alkaloids, some aromatic compounds, macrolides, peptides and terpenes, they act as reducing agents to produce nanoparticles from metal salts without producing any toxic by-product. Once the algal biomolecules are identified, the nanoparticles of desired shape or size may be fabricated. The metal and metal oxide nanoparticles thus synthesized have been investigated for their antimicrobial activity against several gram-positive and gram-negative bacterial strains and fungi. Their dimension is controlled by temperature, incubation time, pH and concentration of the solution. In this review, we have attempted to update the procedure of nanoparticle synthesis from algae, their characterization by UV-vis, Fourier transform infrared spectroscopy, transmission electron microscopy, scanning electron microscopy, x-ray diffraction, energy-dispersive x-ray spectroscopy, dynamic light scattering and application in cutting-edge areas.
KeywordsMetal and Metal Oxide Nanoparticles Controlling Factors Biosynthesis Characterization and Mechanism Antimicrobial Activity
The nanoparticles are the most fundamental component in the fabrication of a nanostructure. Several synthetic routes are used for the fabrication of nanoparticles of diverse morphology and size. Although these procedures have offered superior quality of nanoparticles, better fabrication procedures are yet to be developed. Currently, scientists have focused their attention on the biosynthesis of nanoparticles involving plant, algae, bacteria, fungi and virus containing proteins, amines, aminoacids, phenols, sugars, ketones and aldehydes which act as reducing agents, capping agents and stabilizers for nanoparticles [1–8].
Metal nanoparticles have gained extensive attention due to their efficient antimicrobial activities because they can be safely used in human system to inhibit the growth of pathogens without damaging the normal tissues. The mechanism of antimicrobial activity of nanoparticles has been ascribed to the generation of free radical and subsequent damage of the microbial cell wall leading to their death . Besides, the nanoparticles poison the enzyme of single cell pathogens such as bacteria, fungi and viruses for oxygen intake without harming the human enzymes . Algae may produce nanoparticles from any metal salt by extracellular or intracellular pathways involving biochemicals or enzymes present in them. However, enzymes and reducing substances are known to be the main constituents of microorganisms and fungi for the production of metal nanoparticles from metal salts [7, 28–30].
Engineered nanoparticles of varying size and shape fabricated from various algal species
Singaravelu et al. 
5.42 ± 1.18
Namvar et al. 
Govindaraju et al. 
Uma Suganya et al. 
Spherical and Hexagonal
Rajathi et al. 
Schröfel et al. 
Lirdprapamongkol et al. 
Spherical and Triangular
Senapati et al. 
30 ± 0.25
Spherical and Triangular
Venkatesan et al. 
Spatial array of Self Assembled Structures
Annamalai and Nallamuthu 
Singh et al. 
Mata et al. 
Vijayan et al. 
Govindaraju et al. 
Mubarak Ali et al. 
Spherical and Triangular
Kathiraven et al. 
Prasad et al. 
Round and Rectangular
Barwal et al. 
Vijayan et al. 
Cubical and Hexagonal
Sinha et al. 
Kathiraven et al. 
Abboud et al. 
Azizi et al. 
18 ± 4
Mahdavi et al. 
In this review article, we have discussed the recent advances in nanoparticle fabrication techniques from algae, their characterization by UV-vis, FTIR spectroscopy, TEM, SEM, XRD, AFM, EDX, DLS and application as antimicrobial agents.
Biosynthesis of silver nanoparticles from polysaccharides extracted from four marine algae namely, Pterocladia capillacae, Jania rubins, Ulva faciata and Colpmenia sinusa has been reported . They were found to be spherical with 7–20 nm diameter. Their antibacterial activity has been ascribed to their attachment to bacterial cell wall inhibiting their vital functions.
Fabrication of silver nanoparticles from Sargassum longifolium alga and their microbial activity against several pathogens have been reported . The yellow reaction mixture comprising of AgNO3 and aqueous algal extract turned brown after 1 h. However, the reaction was completed after 32 h, the intensity of which is time dependent. The absorption peak at 440 nm indicated the formation of polydispersed silver nanoparticles. It has been reported that the pH of the reaction mixture exhibited a significant role in the silver nanoparticle synthesis. The colour change of the reaction mixture was slower at low pH 6.2 than that at high pH 8.4. The colour intensity of the reduction process was increased with the increase of the pH. The antifungal activity against Aspergillus fumigatus, Candida albicans and Fusarium sp. was found to increase with increasing concentration of silver nanoparticles .
Biosynthesis and antibacterial activity of silver nanoparticles of 25–44 nm diameter using fresh water green alga, Pithophora oedogonia, has been reported. IR spectrum and quantitative analysis of the extract showed the presence of carbohydrates, saponins, steroids and proteins which reduce AgNO3 to silver nanoparticles. They were found to be more effective against gram negative bacteria than gram positive ones .
Kathiraven et al.  have also reported the biosynthesis of silver nanoparticles from marine alga, Caulerpa racemosa and their antibacterial activity against human pathogens. They were (silver nanoparticles of 5–25 nm) crystalline with face-centred cubic geometry and effective against Staphylococcus aureus and Proteus mirabilis bacteria at a very low concentration (5–15 μL). Silver nanoparticles were synthesized from 14 bacteria and microalgae. It was observed that the nanoparticles were produced by extracellular polysaccharides even in the dark. Spherical, elongated and irregular silver nanoparticles of different dimensions and morphology were obtained which vary from one species to another . The antibacterial activity was tested against six pathogenic bacteria. The mechanism involves free radical formation which causes damage to the cellular membrane.
Small gold nanoparticles of uniform shape with an average size of ~5 nm were obtained from blue green alga, Spirulina platensis . The protein extract of alga and HAuCl4 in a 1:1 ratio in the presence of NaOH was incubated at room temperature for 48 h. Colour change from green to greyish yellow and eventually to ruby red showed the formation of gold nanoparticles . Three distinct peaks at 685, 524 and 385 nm were observed along with an excitation maximum at 620 nm. The peaks at 685 and 629 nm assigned to HOMO and LUMO charge transfer transitions  are the frequencies for secondary amines, OH and COO− groups which would have stabilized the gold nanoparticles. Their antibacterial activity against Bacillus subtilis was examined. The results indicated that nanoparticles caused damage to cells by producing pits in the outer cell wall which disrupt the normal functioning of the bacteria . Since the gold nanoparticles are smaller than the thickness of bacterial cell wall they can easily penetrate into the cell and inhibit their growth.
Extracellular biosynthesis of gold nanoparticles using marine alga Sargassum wightii of 8–12 nm has been reported . The reaction was completed in 15 h with a visibly distinct ruby colour with an absorption maximum at 527 nm. The TEM images showed monodispersed gold nanoparticles, where they are predominantly sphere of 11 nm.
Parial et al.  have reported the fabrication of gold nanoparticles from three cynobacteria (Phormidium valderianum, Phormidium tenue, Microcoleus chthonoplastes) and four green algae (Rhizoclonium fontinale, Ulva intestinalis, Chara zeylanica, Pithophora oedogoniana) at different pH at 20 °C. Generally, the gold nanoparticles were spherical at neutral pH and at pH 9 along with hexagonal and triangular ones. At pH 7 and 9, they exhibited a single absorption between 520 and 534 nm, while at about pH 5, two absorption bands at 520 and one ~600–670 were observed. The peaks vary with pH, concentration of the solution and the nature of cynobacteria and algae. These factors also affect the shape and size of the gold nanoparticles. At pH 5, the small spherical particles (15 nm) together with nano rods (411 × 32 nm) with some larger ones (17 nm) are produced. It is, however, noted that all gold nanoparticles are monodispersed with some aggregation.
Dahoumane and co-workers  have synthesized gold nanoparticles from living cells of Euglena gracilis microalga. The biomaterial in the alga act as reducing agent, capping agent and catalyst similar to other marine algae . The pH, reaction time, temperature and concentration are controlling factors for the nanoparticles yield. It has been proposed that gold nanoparticle formation and release occur in three steps: (1) uptake of Au+3, (2) reduction of Au+3 to Au0 and (3) release of gold nanoparticles into the solvent. They are well dispersed and do not aggregate. They are spherical in shape whose dimensions vary from 10 nm to several 100 nm. AuCl3 concentration of 10−3 M is lethal to E. gracilis which suggests that all algae have a tolerance limit and certain capacity to reduce metal ions to protect themselves from the toxic influence of Au3+/Au0.
Antibacterial activity of gold nanoparticles (modified, )
Positive Control (tetracycline)
Negative Control (chloroauric acid)
Metal Oxide Nanoparticles
Biosynthesis of zinc oxide nanoparticle from aqueous extract of brown marine macroalga, Sargassum muticum has been reported . The colour of the reaction mixture containing ZnO and algal extract changed from dark brown to a pale white colour indicating the synthesis of zinc oxide nanoparticle. Surface and hydroxyl moieties of polysaccharide present in the extract are involved in the formation of zinc oxide nanoparticles of 30–57 nm. They were agglomerated with hexagonal structure. Authors have concluded that the synthesized zinc oxide nanoparticles prepared from S. muticum is expected to have notable applications in pharmaceutical and biomedical fields and in cosmetic industries.
Abboud et al.  have reported the synthesis of copper oxide nanoparticles of 5–45 nm dimension from B. bifurcate algal extract. They were shown to be a mixture of Cu(I) and Cu(II) oxides and were crystalline in nature. Transition metal oxide nanoparticles are an important class of semiconductors and because of incompletely filled d orbitals, they find application in magnetic storage media, energy transformation, electronic and catalysis [59–61]. The formation of copper oxide nanoparticles was confirmed by a change in colour when 1 mM solution of CuSO4 was added to B. bifurcate extract at ambient temperature. Their UV-vis spectra showed distinct change in the absorption peaks owing to the presence of diterpenoids in the extract followed by the formation of cuprous oxide and cupric oxide nanoparticles [62, 63]. The CuSO4 undergoes partial reduction to Cu(I) and Cu(II) oxides which is reflected from the blood red colour exhibiting absorption at 260 and 650 nm. The TEM image showed that majority of the nanoparticles are spherical, although some elongated ones were also observed. Since the nanoparticles are a mixture of cupric oxide and cuprous oxide the XRD pattern showed the presence of two crystalline phases, monoclinic copper(I) oxide and copper(II) oxide with cuprite structure. The antibacterial activity of algal extract and copper oxide nanoparticles was tested against Enterobacter aerogenes and Staphylococcus aureus. It was observed that the algal extract alone was ineffective while copper oxide nanoparticles were significantly active against two bacterial strains.
Iron oxide nanoparticles were synthesized from FeCl3 with an aqueous extract of brown alga Sargassum muticum at 25 °C. The polysaccharides present in the algal extract reduce the FeCl3 to Fe3O4 nanoparticles of 18 ± 4 nm size which are mainly cubic in shape .
Very few reports are available on the toxic effects of several metal nanoparticles on marine organisms including algae, bacteria and protozoa in order to have a data bank for risk assessment . Aruoja et al.  have synthesized Al2O3, Co3O4, CuO, Fe3O4, MgO, Mn3O4, Sb2O3, SiO2, ZnO, TiO2, WO3 and Pd crystalline nanoparticles. They are 8–21 nm in size. Some of these oxide nanoparticles are acidic, some are basic and others are amphoteric in nature. They give stable suspension in water. Their toxicity has also been investigated against one alga (Pseudokirchneriella subcapitata), three bacteria (Vibrio fischeri, Escherichia coli, Staphylococcus aureus) and one protozoa (Tetrahymena thermophila).
Categorization of nanoparticles based on the toxicity values (EC50 or MBC, mg compound l−1) to bacteria, protozoa and algae. All nanoparticles were tested in nominal concentrations from 0.01 up to 100 mg l−1 
EC50 or MBC, mg compound l−1
72 h EC50
24 h EC50
30 min EC50
24 h MBC
24 h MBC
Vibrio fischeri (G−)
Escherichia coli (G−)
Staphylococcus aureus (G+)
2 % NaCl
CuO, ZnO, Pd
Co3O4, Fe3O4, Mn3O4, TiO2
Al2O3, SiO2, WO3
ZnO, Pd, WO3, Sb2O3
Fe3O4, Mn3O4, Pd
Al2O3, Co3O4, MgO, Mn3O4, Pd, Sb2O3, SiO2, WO3
Al2O3, Co3O4, Fe3O4, MgO, Mn3O4, SiO2, TiO2
Al2O3, MgO, Mn3O4, Sb2O3, SiO2, TiO2, WO3
Al2O3, Co3O4, MgO, Sb2O3, SiO2, TiO2, WO
The pH of the suspension containing ZnO and the algae does decrease from 8 to 4, but virtually there is no variation in toxicity as a function of pH . Hartmann et al.  have studied the toxicity of TiO2 nanoparticles of 10, 30 and 300 nm against Pseudokirchneriella subcapitata alga. All the three types of particles exhibited algal growth inhibition. The ecotoxicity of Cd to alga, P. subcapitata, in presence of 2 mg L−1 of TiO2 was reduced probably due to non availability of Cd in presence of TiO2 nanoparticles. The toxicity was also found to be dependent on the nanoparticles and their concentration.
Ji and co-workers  have studied the toxicity of Al2O3, SiO2, ZnO and TiO2 nanoparticles towards green alga, Chlorella sp. Al2O3, SiO2 and TiO2 (DJ3, rutile) did not show significant toxicity although ZnO and TiO2 (HR3, anatase) inhibited the algal growth in 20 and 30 mg L−1 nanoparticles in aqueous solution.
The ecotoxic effects of oxide nanoparticles are dependent on their size and type. Even at very high concentration (1000 mg L−1), the algal growth did not show any variation from the second day to the sixth day of exposure. Nano Al2O3 showed growth promotion at the fourth day by about 19 %. Lin and Xing  have found nano Al2O3 as nontoxic to five plant species. However, at higher concentration of 2000 mg L−1 of Al2O3, root growth is inhibited . Such experimental results may not be applied in the field because such a high concentration is seldom achieved in aquatic system as the algae etc. will dry up due to large accumulation of nanoparticles and other toxic materials.
The toxic effect of the nanoparticle and their bulk material are not the same. For instance, the Chlorella sp. toxicities for different form of Zn follow the order: Zn2+ > nano ZnO > bulk ZnO even when their concentrations are below 50 mg L−1. At higher concentration (>50 mg L−1), the toxicity of ZnO nanoparticles has been shown to be higher than Zn2+. The toxicity also depends on particle size, crystal structure, rutile and anatase. Anatase TiO2 is more toxic than rutile TiO2. Ji et al.  have suggested that anatase TiO2 release larger quantity of ROS than rutile TiO2 resulting in an increase toxicity . However, the toxicity of anatase TiO2 nanoparticles decreases if their size increases above 33 nm . Nano TiO2 and nano ZnO can produce photocatalytic ROS in presence of UV light , but experimental evidences demonstrated larger production of ROS even in the dark . It is therefore concluded that there are other possible reasons for the toxicity of nanoparticles besides the ROS production.
Algae are considered as significant nanofactories and hold a huge potential as ecofriendly and cost-effective tools, avoiding toxic, harsh chemicals and the high energy demand required for physiochemical fabrication. In the present review, we have discussed the biosynthesis of metal and metal oxide nanoparticles from a variety of algae and their toxicity against several pathogenic gram-positive and gram-negative bacterial strains. The proteins, polysaccharides, amines, amino acids, alcohols, pigments, carboxylic acids carbohydrates and sugars have been shown to act as reducing agents. Also, they act as capping and stabilizing agents for the fabricated nanoparticles. The results suggest that the functionalized metal nanoparticles may be exploited in the treatment of infectious diseases caused by bacteria and fungi. They can also be used in phytomining and sequestering metals from waste disposals by redox process.
The authors are thankful to publishers for permission to adopt figures in this review.
AH gathered the research data. AH and KSS analyzed these data findings and wrote this review paper. Both authors read and approved the final manuscript.
The authors declare that they have no competing interests.
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
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