Introduction

The use of science and technology in nanotechnology is to manipulate matter at the molecular level. The properties of matter differ dramatically from their macroscopic bulk properties at the nanoscale level. The ability to design, characterize, produce, and use structures, devices, and systems by manipulating shape and size at the nanoscale scale is known as nanotechnology [1]. The manufacturing of nanoparticles from a range of materials is used in the production of important goods such sophisticated materials, energy storage technologies, electronic and optical displays, and pharmaceuticals that save lives. There are numerous physical and chemical techniques for creating nanoparticles, but more recently, the development of metal nanoparticles utilizing living cells like bacteria, fungi, or plants has caught the attention of scientists [2]. Nanotechnology, which is the science and art of manipulating matter at the atomic or molecular level, has the potential to significantly advance environmental protection technologies [3]. Due to their small size and unique characteristics compared to the bulk version of the same substance, nanoparticles have greatly advanced the disciplines of biosensors, biomedicine, and bio nanotechnology.  In addition to being used for diagnosis, therapeutic medication delivery, and the creation of treatments for a variety of diseases and disorders, nanotechnology is also used in medicine.  The design and development of many other types of unique goods offers a great deal of promise thanks to the tremendous power of nanotechnology, which also has the potential to be used in medicine for the early detection, treatment, and prevention of disease [4]. Since the turn of the century, researchers have been studying nanotechnology. Since Nobel laureate Richard P. Feynman introduced the concept of "nanotechnology" in his well-known 1959 lecture "There's Plenty of Room at the Bottom" (Feynman, 1960), the area of nanotechnology has undergone numerous breakthrough advancements. Materials of all kinds were created at the nanoscale through nanotechnology. Nanoparticles (NPs) are a diverse class of materials that comprise substances that are particulate and have at least one dimension less than 100 nm [5].

Proposed the following definitions for the scientific terms that have been used:

·        Nanotechnology: Manipulation and control of matter on the nanoscale dimension by using scientific knowledge of various industrial and biomedical applications.

·        Nanoparticle: Nano-object with three external nanoscale dimensions. The terms nanorod or nanoplate are employed, instead of nanoparticle (NP) when the longest and the shortest axes lengths of a nano-object are different.

·        Nanoscale: Approximately 1 to 1000 nm size range.

·        Nanomaterial: Material with any internal or external structures on the nanoscale dimension.

·        Nanoscience: The science and study of matter at the nanoscale that deals with understanding their size and structure-dependent properties and compares the emergence of individual atoms or molecules or bulk material related differences [6].

The goal of the ecologically friendly process known as "green synthesis" is to reduce energy use, remove hazardous waste, and use ecological solvents like water, ethanol, and ethyl acetate. For the purpose of creating metal/metal oxide nanoparticles using plant leaf extracts, many Researchers have used green synthesis techniques. Plants have biomolecules with an exceptional ability to transform metal salts into nanoparticles, such as carbohydrates, proteins, and coenzymes. Traditional chemical synthesis is less advantageous than green synthesis since it is more expensive, produces less pollution, and protects both the environment and human health.

Manganese Iron Oxide Nanoparticles are high surface area particles with typical sizes of 5 to 100 nm and specific surface areas (SSA) in the 25 to 50 m²/g  range. Nano Manganese Iron Oxide Particles are also offered in Ultra High Purity and High Purity, as well as coated and dispersed forms. Iron Manganese oxide’s compound formula is Fe₂MnO₄ and their Molecular weight is 230.63. Its appearance is brown to black powder and melting point is >300ºC. Iron (atomic symbol: Fe, atomic no: 26) is a Block D, Group8. Manganese (atomic symbol: Mn, atomic number: 25) is a Block D, Group 7 [7]. The excellent physico-chemical properties of manganese oxide nanoparticles can be used in a variety of disciplines, including water purification, optoelectronics, molecular sieves, batteries, and magnetic materials. The primary focus of future research in solar cells will be on syntheticmanganese oxide nanoparticles that are being examined for their optical and electrical properties [8]. Manganese oxide nanoparticles have remarkable physico-chemical properties that make them useful in many fields, including water purification, optoelectronics, molecular sieves, batteries, and magnetic materials. Future studies on solar cells will mostly concentrate on artificial manganese oxide nanoparticles, whose optical and electrical characteristics are being scrutinized [9]. Different formulations of Super paramagnetic iron oxide nanoparticles have been commercialized to date for use as contrast agents in magnetic resonance imaging (MRI) [10].

II. Material and methods

A.     Selection of Fungi species

Aspergillus species [Aspergillus Niger] is isolated from microbial culture of Siddhachalam Laboratory Raipur. This species is used as pattern for the synthesis of Fe₂MnO₄ nanoparticle.

B.     Preparation of culture media for fungi

For 250 ml - 9.75 gm of high media of Potato Dextrose Agar Media, 250 ml of distilled water, 2 gm of Agar.     

C.     Isolation and observation of Fungi

High Media Potato Dextrose Agar Media [PDA] and Agar were added in distilled water and mixed homogenise it, and then after autoclaved, and under laminar air flow the media will be poured and inoculated by fungi from inoculating loop in sterile petri plate. Poured   plates were incubated at 37 c in incubator for 24-48 hours, and fungal species [Aspergillus] was observed.

D.     Preparation of biomass

The isolated Aspergillus spp. was inoculated in a centrifuge tube [falcon tube or conical tube] with 10 ml of distilled water and incubated for 2 hours in rotary shaker at 27◦C. After incubation, the fungus was harvested by centrifugation. The pellet was washed with 10 ml of triple distilled water under sterile condition and was used for the synthesis of Fe₂MnO₄.

E.     Synthesis of Fe₂MnO₄ Nanoparticles

Chemical composition for synthesis of Fe₂MnO_4.

1. Ferrous Sulphate  -  2.7802 gm  

2. Manganese Sulphate - 1.6902 gm

3. Ethyl Glycol

4. Fungus

About 100 ml of distilled water was taken in a conical flask and all the chemical composition was added to it. Then it will be keep in hot plate until it dissolve, and autoclave for 30 min, then after centrifuge it. It was centrifuged and the condensed precipitate was washed with distilled water and finally washed with alcohol and put into the hot air oven for 4 hours and then it was put into the muffle furnace for 4 hours and got the crystalline particles.

1.      UV-Vis spectra analysis

Before calcination of biogenic Fe₂MnO₄ particles were characterized by UV-Visible Spectrophotometer. Before putting the sample in the UV-Visible Spectrophotometer, it is diluted with water or Ethylene Glycol because Fe₂MnO₄ Nanoparticles is soluble in Ethylene Glycol, the sample was directly placed in Spectrophotometer due to which the absorption of Fe₂MnO₄ Nanoparticles was recorded in the transmittance mode in the region of 200-300 by UV visible light and the spectrum of the sample is visible on the screen which is displayed as graph.

2.      FT-IR

The chemical composition of the synthesized Iron Manganese oxide nanoparticles was study by using FT-IR spectrometer (Perkin elemer LS-65 Luminescence spectrometer).

3.      Methylene Blue Dye Degradation       

Because Fe₂MnO₄ is soluble in water, the Fe₂MnO₄ sample solution with distilled water was used for the Methylene Blue Dye Degradation Analysis. Using UV-Vis Spectroscopy, a different absorption graph was obtained for each cycle to demonstrate the degradation of Methylene Blue by metal oxide over time.

4.      For Antimicrobial study

Well diffusion method is used for the antimicrobial testing of Fe₂MnO₄ NPs. Well diffusion method is widely used for antimicrobial testing of antibiotics and other metal compound, this method is also used to obtain the minimal inhibitory concentration of a compound. In this method, the microorganism to be tested is spreaded and small wells are formed through the borer and are filled with the solution of metal oxide nanoparticles and left for diffusion of the nanomaterials. The NPs shows the effect    inhibiting the growth of the cultured organisms.

4.1 For Antibacterial Study

 NAM media should be prepared for antibacterial study of Fe₂MnO₄ NPs.

Media is prepared, poured into petriplates and well were formed, pure culture, of a bacterial colony inoculated by spreading and well formed .Little amount of Fe₂MnO₄ NPs solution is added to the well and incubated at 37ºc for overnight and observed.

4.2 For Antifungal Study

PDA media should be prepared for antifungal study of Fe₂MnO₄ NPs.

Media is prepared, poured into Petri plates and well were formed, pure culture of a fungal colony is inoculated by spreading and well formed. Little amount of Fe₂MnO₄ NPs solution is added to the well and incubated at 37for 48 hours and observed.

III. Result and Discussion

A.     UV-Vis Spectra analysis

The UV-Vis spectrophotometer measures the UV-Vis spectrum of produced NPs. The band gap of the synthesized Fe₂MnO₄ NPs was computed from UV-Vis spectra, and it was determined to be 3.0. We obtained the absorbance of the created Fe₂MnO₄ NPs at point. We deduced from UV absorbance spectra that Fe₂MnO₄ reacts with UV light and functions as a photo catalyst to break strong covalent bonds.

Fig: 1 Showing result UV-Vis Spectrophotometer of Fe₂MnO₄ NPs synthesized from (Aspergillus niger) fungi

We apply the Touc Relation, which is provided by the equation, for band gap;

αhv = A (hv – Eg)n

Where,

α = Absorption Coefficient

hv = Incident Wavelength

Fromthe UV-Vis spectrophotometer measures the UV-Vis spectrum of produced NPs. The band gap of the synthesized Fe₂MnO₄ NPs was computed from UV-Vis spectra, and it was determined to be 3.0.

Graph calculate the band gap:

= 3.0

Fig: 2 Band gap of Fe₂MnO₄ NPs synthesized from (Aspergillus niger) fungi

B.     FT-IR

To examine the functional groups ofFe₂MnO₄ NPs, FT-IR spectra is used. A FT-IR spectrophotometer was used to acquire the absorption spectrum in the region. The  FTIR spectrum of the Fe₂MnO₄ nanoparticles before and after heating process. It is observed that the bands at 1129.32 and 962.80 were assigned to the bending vibrations of primary and secondary amines and carboxylic groups respectively.

Fig: 3 FTIR of Fe₂MnO₄ NPs synthesized from (Aspergillus niger) fungi

C.     Methylene Blue Degradation Analysis

The chemical substance known as methylene blue has the molecular formula C₁₆H₁₈C₁N₃S. Fe₂MnO₄ NPs were used as a catalyst to degrade Methylene blue, and the UV-Visible spectroscopic method was used to measure the degree of the deterioration. Methylene blue dye's absorption peaks inwater were discovered to be centered at 664 nm in the visible spectrum.

Figure 4: Showing Methylene blue dye degradation

Methylene Blue Dye degradation was used to analyze the photocatalytic activity of synthetic Fe₂MnO₄ NPs at regular intervals. For the test, a small amount of Fe₂MnO₄ NPs sample solution was introduced to the MB solution along with 10-1 Methylene blue dye. The reduction in dye and photocatalytic effect is shown by the change in color of MB from deep to light blue. The sample data is recorded by UV-Vis Spectrophotometer for around 40 cycles in a regular interval of time, and the graph is displayed. The photo catalytic activity of Fe₂MnO₄ NPs is demonstrated by a decrease in absorbance at various wavelengths over time.

Fig:4 Fe₂MnO₄ Photo catalytic Absorbance graph.

D.     Antimicrobial Study

For the Antibacterial test, solid agar medium is used, the well diffusion method is used. In the bacterial culture plate, the well containing Fe₂MnO₄ NPs is not showing a zone of inhibition and in the fungal culture plate also not showing zone of inhibition.

(A)                              (B)

Fig: 5 (A) Fe₂MnO₄ Antibacterial test, Staphylobacillus

(B)   Fe₂MnO₄ Antifungal test, Alternaria

  IV. CONCLUSION