BIOLOGICAL APPROACH OF MAGNESIUM OXIDE NANOPARTICLES SYNTHESIZE BY SPIRULINA PLATENSIS

World Journal of Pharmaceutical Research Mohammad et al.

World Journal of Pharmaceutical Research SJIF Impact Factor 5.990

Volume 4, Issue 7, 1234-1241.

Research Article

ISSN 2277– 7105

BIOLOGICAL APPROACH OF MAGNESIUM OXIDE NANOPARTICLES SYNTHESIZE BY SPIRULINA PLATENSIS Mohammad Irfan Ali1*, Gaurav Sharma1, Manoj Kumar2, Nakuleshwar Dut Jasuja1, Rajgovind1 1 2

School of Sciences, Suresh Gyan Vihar University, Rajasthan, India.

Marine Biotechnology Laboratory, Department of Botany, University of Delhi, North Campus, Delhi- 110007, India. ABSTRACT

Article Received on 24 April 2015, Revised on 18 May 2015, Accepted on 09 June 2015

Background-The present study explores extracellular synthesis of magnesium oxide nanoparticles (MgO NPs) using the aqueous extract of Spirulina platensis. Biosynthesized MgO NPs were characterized by UV-Vis spectroscopy, XRD and FTIR studies and finally tested for

*Correspondence for

antibacterial activity. Methods- Biosynthesis using extract of S.

Author

platensis showed the formation of well scattered, highly stable,

Mohammad Irfan Ali

spherical MgO NPs with an average size of 30-40 nm. The size and

School of Sciences,

morphology of the nanoparticles were confirmed by SEM and TEM

Suresh Gyan Vihar University, Rajasthan,

analysis. Results- Furthermore, the synthesized nanoparticles exhibited good antibacterial activity against pathogenic gram-negative i.e.

India.

Escherichia Coli- MTCC-9721, Proteus vulgaris- MTCC-7299, Klebsiella pneumonia- MTCC-9751 and gram-positive i.e. Staphylococcus aureus- MTCC9542, S. epidermidis- MTCC- 2639, Bacillus cereus- MTCC-9017 bacteria. The MgO NPs had shown maximum zone of inhibition (ZOI) i.e. 25.3±0.48 in S. aureus- MTCC-9542. Conclusion- Use of such a biological method provides a simple, cost-effective alternative model for the synthesis of nanomaterials in a large scale that could be exploit in biomedical applications. KEYWORDS: Magnesium oxide nanoparticles, Spirulina platensis, Antibacterial activity, and Biosynthesis.

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INTRODUCTION Nanotechnology have emerged over the past decade as the forefront of science and technologies.[1] The intrinsic properties of nanoparticle are mainly determined by size, shape, composition and crystallinity. Nanoparticles provide a narrow size distribution, which is required to obtain a uniform response of material.[2] Metal oxides such as ZnO, MgO, CaO and TiO2 are of particular interest through not only stable under harsh process conditions, however generally regarded as safe materials to human beings.[3] Metal oxide are toxic to microbes at very low concentrations and they kill microbes disrupting permeability of cell membrane by binding to intracellular proteins and inactivating them.[4] Although the exact antibacterial mechanism of MgO NPs is not clear, three main antibacterial mechanisms have been projected, such as the formation of reactive oxygen species (ROS), the interaction of MgO NPs with bacteria, subsequently destruction of the bacterial cell, and an alkaline effect. The nano-crystalline metal oxide powders have apprehended consideration of scientists due to their potential applications in adsorption,[5] catalysis,[6] sensors and dye-sensitized solar cell.[7] It has been reported that the shape and size of magnesium oxide nanoparticles provide them high specific surface and reactivity, because of the high concentration of edge and corner on their surface.[8, 9] Previously, phyto-reduction method for the synthesis of metallic nanoparticle have been successfully exploited.[10, 11, 12] The present study was carried out with the main objective of evolving a simple method for the extracellular synthesis of MgO nanoparticles by S. platensis. MgO nanoparticles are characterised by FTIR, XRD TEM and SEM analysis. Antibacterial studies of MgO nanoparticles were observed against pathogenic gram-negative i.e. Escherichia Coli- MTCC-9721, Proteus vulgaris- MTCC-7299, Klebsiella pneumonia- MTCC-9751 and gram-positive i.e. Staphylococcus aureus- MTCC-9542, S. epidermidis- MTCC- 2639, Bacillus cereus- MTCC-9017 bacteria. MATERIALS AND METHODS Microorganism and Culture condition The experimental organism S. platensis was isolated from Jalmahal, Jaipur, Rajasthan (India) and cultivated in Zarrouk’s medium under different temperature and illuminated with white fluorescent lamps at a light intensity of 2,000 lux.[13] Preparation of microalgal Extract Typically, 5 g (dry weight) S. platensis biomass was suspended in 100 ml of double distilled sterile water for 15 min at 100°C in an Erlenmeyer flask. After boiling, the mixture was

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cooled and centrifuged at 10,000 rpm for 15 min. Supernatant was collected and was stored at 4 ºC for further analysis. Biosynthesis of Magnesium oxide nanoparticles In the typically synthesis process of Magnesium oxide nanoparticles (MgO NPs), add 2 ml of pure aqueous extract of S. Platensis drop wise into the 100 ml of 1 mM of Magnesium nitrate (MgNO3) solution in 250 ml conical flask. The reaction mixture was kept at 100°C under constant mechanically stirring for 3 hours and allowed to settle for 24 hours. At that time the supernatant was discarded carefully and the remaining solution was centrifuged at 10,000 rpm for 15 minutes. The obtained magnesium oxide nanoparticles (MgO NPs) was washed three times using distilled water to remove the impurities. Characterization of prepared nanoparticles The Characterization of MgO NPs was carried out by surface plasmon resonance band using a UV-Visible Spectroscopy 1800 of Shimadzu, Kyoto, Japan. Micrograph of MgO NPs was obtained by scanning electron microscope of S-4500, Hitachi, Chiyoda-ku, Japan. TEM micrograph of the MgO NPs was observed using the TEM instrument of JEOL JSM 100cx. TEM device conducted at an increasing voltage of 200kv. The FTIR spectrum was recorded on a Perkin Elmer FTIR system Spectrum GX model. All measurements were carried out in the range of 400 – 4,000 cm-1 at a resolution of 4 cm-1. Antibacterial activity of MgO NPs A turbid liquid sample of each bacterial strain with an OD of McFarland of 0.5 (1×108 CFU/mL) was prepared in an isotonic NaCl (0.85%) solution. Furthermore, this solution was diluted ten times (1×107 CFU/mL) and used as inoculums. The MHA plates were inoculate with test disk and Gentamicin acquired by Hi-Media, Mumbai, used as control. The zone of inhibition (ZOI) observed at the surrounding area of MgO NPs solution after incubation at 37°C for 24 hours. The experiments were done four replicate and mean values of ZOI were reported. The MgO NPs prepared by S. platensis were used to evaluate antibacterial activity against Gram (-) and Gram (+) Bacteria (Escherichia Coli- MTCC-9721, Proteus vulgaris- MTCC7299, Klebsiella pneumonia- MTCC-9751, Staphylococcus aureus- MTCC-9542, S. epidermidid- MTCC- 2639,Bacillus cereus- MTCC-9017,) on MHA plates by Kirby- Bauer disk diffusion method.[14]

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RESULTS AND DISCUSSION In this study, extracellular synthesis of MgO NPs has been shown by the aqueous extracts of S. platensis. These extract when interacted with the Magnesium Nitrate solution forms a light yellow colour solution due to the reduction of the magnesium ion to magnesium hydroxide followed by transformation of magnesium hydroxide to MgO NPs. (Figure 1). UV–Visible spectrum recorded from the MgO NPs solution showing surface plasmon absorption bands at 256.5 nm indicates the presence of MgO NPs (Figure 2). The morphological characteristic of biosynthesised MgO NPs were studied by scanning electron microscope, using an instrument of Hitachi S-4500. The SEM images showed that most of the particles are spherical in shape and do not create big agglomerates which indicates that they were in the direct contact, but stabilized by a capping agent (Figure 3a). The TEM images revealed that MgO NPs in the range of 30-40 nm (Figure 3b).

Figure 1: The pictures show the (a) S. platensis extracts, (b) MgNO3 solution and (c) MgO NPs solution.

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Figure 2: UV–Visible spectrum recorded from the MgO NPs synthesized by S. Platensis.

Figure 3(a) The SEM images (b) TEM image of MgO NPs synthesized by S. Platensis Antibacterial activity of magnesium nanoparticles The antibacterial activity of nano-crystalline MgO was evaluated on Gram negative and Gram positive bacteria. Table-1, shows the four replicates experiments of zone of inhibition (mm) around the disc with cell free aqueous extracts mediated synthesized silver MgO NPs. The study revealed that MgO NPs (50 µg/100 µL) had shown maximum inhibitory effect against Staphylococcus aureus- MTCC-9542 i.e. 25.3±0.48 followed by S. epidermidisMTCC- 2639 (23.8±0.63), Klebsiella pneumonia- MTCC-9751 (21.8±0.85), Bacillus cereuswww.wjpr.net

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MTCC-9017 (20.8±0.48), Proteus vulgaris- MTCC-7299 (20.5±0.65) and Escherichia ColiMTCC-9721 (18.8±1.31). (Figure 4a-f). These result correlate with findings of Tang et al.[15] The antibacterial activity of MgO NPs is due to disrupt cell wall. This is caused by ROS (reactive oxygen species) i.e. active superoxide ions are generated on the surface of the oxide, which react with the carbonyl group of peptide linkages of cell wall leading to degradation of the proteins. In case of Nano-crystalline MgO the surface area increases, which determines the potential number of reactive groups on the particle surface, ultimately enhance O2concentration in solution, which are expected to show high antibacterial activity due to more effective disruption of the cell wall of the bacteria.[16, 17] When the cell walls are disrupted, leakage of metabolites occurs and the cell functions are stopped, thus preventing the organism from reproducing and other functioning.

Figure 4(a-f) Antibacterial Activities of on (a) Klebsiella pneumonia (b) Staphylococcus epidermidis (c) Staphylococcus aureus (d) Bacillus cereus (e) Proteus vulgaris (f) Escherichia Coli of MgO NPs (50 µg/100 µL) synthesized by S. platensis. Table 1: Antibacterial activity of MgO NPs (Zone of Inhibition (ZOI) in mm) Bacterial strain Escherichia Coli- MTCC-9721 Proteus vulgaris- MTCC-7299 Klebsiella pneumonia- MTCC-9751 Staphylococcus aureus- MTCC-9542 S. epidermidis-MTCC- 2639 Bacillus cereus- MTCC-9017

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ZOI-1 21 22 24 25 24 22

ZOI-2 19 21 22 24 25 20

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ZOI-3 15 19 21 26 24 20

ZOI-4 20 20 20 26 22 21

Mean±SE 18.8±1.31 20.5±0.65 21.8±0.85 25.3±0.48 23.8±0.63 20.80.48

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CONCLUSION MgO NPs were prepared by a biological method in this paper. This biological method approach toward the bio-synthesis of MgO NPs has numerous benefits i.e. non-toxic, cost effective, rapid reduction, economic viability. The results suggest that biosynthesized Nanocrystalline MgO are a promising bactericidal agent due to their high resistance to harsh processing conditions. Nano-crystalline MgO from the S. Platensis has good antibacterial potential against both Gram-negative and Gram-positive bacteria. This biosynthesis method can be a promising method for the preparation of other metals and metal oxide nanoparticles and can be valuable in environmental, biotechnological, pharmaceutical and medical applications. Future prospects of this research would be to large scale production of MgO NPs using S. platensis and to ascertain its efficacy against extensive spectrum of microbial population. Further investigations would involve discovering the potency of S. platensis to synthesize magnesium nanoparticles. ACKNOWLEDGMENTS The authors are sincerely thankful to Mr. Sunil Sharma, Chancellor and Dr. Sudhanshu Sharma, Chief Mentor of Suresh GyanVihar University for providing a platform for this research. The authors also appreciation vows to USIC, University of Rajasthan, Jaipur, India for providing SEM and TEM facilities. REFERENCES 1. Rajendran R, Balakumar C, Jayakumar S, Rajesh EM, Use of zinc oxide nano particles for production of antimicrobial textiles Int. Jour. Engn. Sci. Tech., 2010; 2: 202. 2. Dickson RM and Lyon LA. “Unidirectional plasmon propagation in metallic nanowires.” J. Phys. Chem. B, 2000; 104(26): 6095–6098. 3. Stoimenov PK, Klinger RL, Marchin GL, Klabunde KJ. Metal Oxide Nanoparticles as Bactericidal Agents. Langmuir, 2002; 18: 6679. 4. McDonnell G, Russell AD. Antiseptics and Disinfectants: Activity, Action, and Resistance. Clin. Microbiol.Rev, 1999; 12: 147- 179. 5. Anupam K, Dutta S, Bhattacharjee C, Datta S. Optimisation of adsorption efficiency for reactive red 198 removal from wastewater over TiO2 using response surface methodology. Can. J. Chem. Eng, 2011; 89: 1274–1280 6. Velmurugan R, Krishnakumar B, Kumar R, Swaminathan M. Solar active nano-TiO2 for mineralization of Reactive Red 120 and Trypan Blue. Arab. J. Chem, 2012; 5: 447–4527.

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Mohammad et al.

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Al-Owais AA, Synthesis and magnetic properties of hexagonally packed ZnO nanorods, 2013; Arab. J. Chem. 6: 229–234. 7. 7Al-Owais, AA. Synthesis and magnetic properties of hexagonally packed ZnO nanorods. Arab. J. Chem, 2013; 6: 229–234. 8. Rajagopalan S, Koper S, Decker S, Klabunde KJ2 Nanocrystalline Metal Oxides as Destructive Adsorbents for Organophosphorus Compounds at Ambient Temperatures. J.Eur.Chem, 2002; 8: 2602. 9. Sharama G, Jasuja ND, Ali MI, Joshi SC. A Review on Nanomedicinal and Nanosensing Potential of Nanoparticles. Int. J. Biol. Chem, 2014, 8(2): 58-84. 10. Sharama G, Jasuja ND, Rajgovind, Prerna S, Joshi SC. Synthesis, Characterization and Antimicrobial Activity of Abelia grandiflora Assisted AgNPs. J Microb Biochem Technol, 2014; 6(5): 274-273. 11. Rajgovind, Sharma G, Jasuja ND, Ali MI, Gupta DK, Joshi SC. Probing Antibacterial Properties of Sida cardifolia Sponsored Silver Nanoparticles International Journal of Recent Research Aspects ISSN: 2349 – 7688, 2015; (Communicated) 12. Rajgovind, Sharma G,

Jasuja ND, Deepak Kr. Gupta DK, Joshi SC. Pterocarpus

marsupium Derived Phyto-Synthesis of Copper Oxide Nanoparticles and their Antimicrobial Activities. J Microb Biochem Technol, 2015; 7: 140-144. 13. Sharma G, Kumar M, Ali MI, Jasuja ND, Effect of Carbon content, Salinity and pH on Spirulina platensis for Phycocyanin, Allophycocyanin and Phycoerythrin accumulation, Journal of microbial and biochemical technology, 2014; 6(4): 202-206. 14. Bauer AW, Kirby WMM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. Amer. I. C/in. Pathol, 1966; 45:493-496. 15. Tang Z, Fang X, Zhang Z, Zhou T, Zhang Z, Shi L, Nanosize Mgo as antibacterial agent: preparation and characteristics. Brazilian jouranal of chemical engineering, 2012; 29(4): 775 – 781. 16. Makhluf S, Dror R, Nitzan Y, Abramovich Y, Jelinek R, Gedanken A. Microwave-assisted synthesis of nanocrystalline MgO and its use as bacteriocide. Adv. Funct. Mater, 2005; 15: 1708-1715. 17. Pal S, Tak YK, Song JM, Does the antibacterial activity of silver nanoparticles depend on the shape of the nanoparticles? A study of the gramnegative bacterium Escherichia coli. Applied and Environmental Microbiology, 2007; 73: 1712-1720.

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