A Study of Iron Oxide Nanoparticles Synthesis by Using Bacteria

Abstract

<jats:p>Nanotechnology is a multi-disciplinary kind of science that covers many areas of scientific techniques, like biomedical, pharmaceutical, agricultural, environmental, materials, general chemistry, general physics, electronics, data sciences and technology, etc.1-4 Nanotechnology is become applied now in the pharmaceutical industry, medicine, electronics, robotics, and tissue engineering. The usage of nanomaterials in the enhancement of delivery systems for various molecules, like DNA, RNA, plasmids, and proteins it is very important today and has been considered widely throughout the last years.2 Nanoparticles have been used to deliver drugs to target tissues and to increase stability against degradation by enzymes.3 Their exclusive size-dependent properties make these materials indispensable and superior in many areas of human activities.3,4 Green synthesis methods are eco-friendly approaches and compatible with pharmaceutical and other biomedical applications, as the toxic chemicals are not used in these methods.5 Iron oxide nanoparticles are most suitable for biomedical applications due to their proven biocompatibility. These particles have an ability to interact with various biological molecules in different ways due to their superparamagnetic properties, high specific area and wide choice of surface functionalization.6 The potential of drug delivery systems based on the use of nano- and microparticles stems from significant advantages such as;7,8 The ability to target specific locations in the body. The reduction of the drug quantity needed to attain a particular concentration approximately the target. The reduction of the concentration of the drug at non-target sites minimizing severe side effects. Living microorganisms, especially Bacillus sp. have a remarkable ability to form exquisite inorganic structures often in nano-dimensions. The development of these eco-friendly methods for the fabrication of nanoparticles is developing into an important division of nanotechnology, especially iron oxide nanoparticles.7,9 Microbes play direct or indirect roles in several biological activities. So use them in the biosynthesis of nanoparticles is a more demanding approach for the bio-production of nanoparticles via a highly stable, eco-friendly process with no toxic chemical and large scale production.10 Our study aims to investigate and detect iron oxide nanoparticles produced by Bacillusims to Sp. Bacteria. METHODOLOGY Bacillus Identification Large gram-positive rods, often in pairs or chains with rounded or square ends (which may have a single endospore). Some species may be Gram variable. The identification was done by using spore stain method, which used to stain the spores of Bacillus species. Spores were in light green, and vegetative cell walls were pick up the counterstain safranin. The media used with conditions ware blood agar incubated in air/CO2 at 35°C-37°C for 24 – 48-hour.11 ABSTRACT The biosynthesis of nanoparticles by using microorganisms is developing as an ecofriendly method for nanoparticle synthesis because of its cheap, simple and non-toxic. Bacillus sp. can be used for producing iron oxide nanoparticles. In addition, it has the ability for the biosynthesis of Fe3O4 nanoparticles. The nanoparticles producing was evaluated by using Ultra Violate-Visible (UV-Visible) and Fourier-transform infrared spectroscopy (FT-IR) methods also the production and size of the nanoparticle was confirmed by X-ray Diffraction and Field Emission Scanning Electron Microscope (FESEM) to confirm the accuracy of iron oxide nanoparticles. pH, Temperature, and Incubation time of production of iron oxide nano-particle also studied.</jats:p>

Green Synthesis: An Eco-friendly Route for the Synthesis of Iron Oxide Nanoparticles

“…Among them, temperature, pH and incubation time for the microbial culture are the paramount factors, as this green approach of synthesis is regulated by the enzymatic system for reduction of metal ions (NADH reductase), so biosynthesis depends upon optimum conditions. High temperature and pH completely inactivates the enzymes and halts their action, in turn, slows down the nanoparticles formation (Sundaram et al, 2012;Garg et al, 2020;Jubran et al, 2020). Besides enzymes, proteins, peptides, cofactors and organic molecules also contributes in the synthesis process as stabilizer, capping and reducing agents (Singh et al, 2016).…”

“…Iron oxide nanoparticles synthesis generally begins with the selection of bacteria (from soil, lyophilized culture). Isolation step includes growing the sample on the media like BH1 agar (Sundaram et al, 2012), Blood agar (Jubran et al, 2020) or nutrient agar (Fatemi et al, 2018) followed by their incubation under controlled conditions at 35-37°C for 24-48 h and in case of blood agar medium incubation is carried out under controlled conditions of carbon dioxide. After the achievement of sufficient growth single colony is transferred to the nutrient medium, such as Luria broth, Nutrient broth or MRS broth (Sundaram et al, 2012;Fani et al, 2018;Fatemi et al, 2018;Torabian et al, 2018;Jubran et al, 2020) followed by incubation and centrifugation to pellet out the bacterial biomass.…”

“…Isolation step includes growing the sample on the media like BH1 agar (Sundaram et al, 2012), Blood agar (Jubran et al, 2020) or nutrient agar (Fatemi et al, 2018) followed by their incubation under controlled conditions at 35-37°C for 24-48 h and in case of blood agar medium incubation is carried out under controlled conditions of carbon dioxide. After the achievement of sufficient growth single colony is transferred to the nutrient medium, such as Luria broth, Nutrient broth or MRS broth (Sundaram et al, 2012;Fani et al, 2018;Fatemi et al, 2018;Torabian et al, 2018;Jubran et al, 2020) followed by incubation and centrifugation to pellet out the bacterial biomass. After centrifugation most critical step of IONPs synthesis initiates by taking either supernatant (Sundaram et al, 2012;Fatemi et al, 2018;Jubran et al, 2020) or cell pellet (Fani et al, 2018;Torabian et al, 2018).…”

“…After the achievement of sufficient growth single colony is transferred to the nutrient medium, such as Luria broth, Nutrient broth or MRS broth (Sundaram et al, 2012;Fani et al, 2018;Fatemi et al, 2018;Torabian et al, 2018;Jubran et al, 2020) followed by incubation and centrifugation to pellet out the bacterial biomass. After centrifugation most critical step of IONPs synthesis initiates by taking either supernatant (Sundaram et al, 2012;Fatemi et al, 2018;Jubran et al, 2020) or cell pellet (Fani et al, 2018;Torabian et al, 2018). The protocol after using pellet (dissolved in Phosphate Buffer Saline) required additional step of centrifugation at 3000 g for 10 min and washing including short incubation in liquid nitrogen (−196°C) for 5 min, followed by subjecting the cells to a brief heat shock by steam bathing the cell pellet at 37°C for 15 min.…”

“…One another astonishing feature of these particles is their potential to join together with different biological molecules like peptides, enzymes, nucleic acids, lipids, fatty acids and various metabolites. Molecules immobilized on the nanoparticles demonstrate superior properties and high reusuability over the unimmobilized counterpart (Singh et al, 2016;Jubran et al, 2020). Different chemical, physical and biological methods have been employed for the synthesis of IONPs.…”