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Author(s): Heena Nayak1*1, Pragya Kulkarni2



    Department of Microbiology, Durg (C.G.), India 492001 Govt. V.Y.T. P.G. Autonomous college

Published In:   Volume - 4,      Issue - 1,     Year - 2024

Cite this article:
Heena Nayak1*, Pragya Kulkarni1 (2024), Industrial algae mediated development and evaluation of Titanium Oxide nanoparticles, their ability to fight bacteria, and environmental application, Spectrum of Emerging Sciences, 4 (1) 2024, 7-12, 10.55878/SES2024-4-1-2

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Nanotechnologies are an area of study and technology that has grown dramatically in the previous few decades due to its importance in many other sectors [1]. The area of nanotechnology in science holds great promise for use in the medical field. Nanotechnology is crucial to current research because it is the most useful technology that can be applied in practically every field, including cosmetics, pharmaceuticals, environmental health, food and feed, chemical industry, agricultural science, energy sector, drug and gene delivery, mechanics, and space industry. Thus, in nanotechnology, it is becoming increasingly vital to use microorganisms, enzymes, and plant extracts in clean, biocompatible, non-toxic, and ecologically acceptable ways [2]. The rapidly emerging discipline of nanotechnology has seen an explosion of attention in recent years. The tiny billionth of a meter is referred to as a "nano" in this context. Numerous applications in the domains of energy, nutrition, and medicine have already made use of these nanostructures' smaller size and distinctive surface chemistry. [3]. Particles with a size range of 1–100 nm are known as nanoparticles (NPs), and they are the primary focus of the emerging field of nanotechnology[4]. In addition to being employed in many chemical and medicinal applications, nanoparticles are also used in everyday life. Generally speaking, there are two types of nanoparticles: inorganic (such metallic and magnetic nanoparticles) and organic (like liposome, chitosan, and micelles)[5]. In addition to being employed in many chemical and medicinal applications, nanoparticles are also used in everyday life. Generally speaking, there are two types of nanoparticles: inorganic (such metallic and magnetic nanoparticles) and organic (like liposome, chitosan, and micelles) [6].

1.    Nanoscale: A range of sizes approximately 1–1000 nm.

2.    Nanoscience:The study of matter at the nanoscale, which looks at how bulk materials change or emerge as individual atoms or molecules, with an emphasis on understanding aspects that depend on size and structure.

3.    Nanotechnology: The scientific investigation of many applications in industry and biomedicine for the manipulation and control of materials at the nanoscale.

4.    Nanomaterial: Nanomaterials are defined as materials having any external or interior features on the nanoscale dimension.

5.    Nano-object: A nano-object is a substance having one or more peripheral nanoscale dimensions.

6.    Nanoparticle: An external nano-object with three small dimensions. In times where the lengths of a nano-object's longest and shortest axes differ, the terms "nanorod" or "nanoplate" are employed rather than "nanoparticles" (NPs).

7.    Nanofiber: A material with three dimensions—two of which are the same externally and one of which is larger—is called a nanofiber.

8.    Nanocomposite: A multiphase structure that contains at least one dimension at the nanoscale.

9.    Nanostructure: A group of connected nanoscale component parts.

10.Nanostructured materials: materials with externally or internal nanostructure [7].

Within material science, green synthesis has become a stable, durable, and sustainable process for creating a variety of nanomaterials, such as metal oxides, hybrids, and materials inspired by biological processes [8]. Recently, scientists have been more interested in the environmentally friendly synthesis of nanoparticles using microbes and plant extracts. The biogenesis of nanoparticles is considered preferable to chemical synthesis due to the creation of hazardous chemical species that are adsorbed on the particle surface during chemical synthesis. This synthesis can involve bacteria, plants, algae, fungi and other living organisms. Many nonoparticles can be produced because of the phytochemical in its extract, which acts as a stabilizing and reducing agent [9]. In addition to possessing exceptional chemical stability and photocatalytic activity, titanium oxide (TiO2) is inexpensive and safe for biological usage. The two types of titanium oxide (TiO2) that make up most of the material are crystalline and amorphous. When particles are in the crystalline state, they are grouped in a regular and comparable manner. The most stable crystalline phase is formed in rutile when the temperature reaches a high of 800ºC. In contrast, the amorphous form’s uneven shape results from the particles haphazard arrangement. The process of depositing thin sheets at 350ºC results in the formation of anatase, an amorphous material [10]. Three polymorphic forms of titanium dioxide with crystalline structure that can be found in nature are rutile, anatase and brookite. The gemstone industry uses these materials extensively [11]. Researched, developed and integrated CuO, ZnO and CeMo nanoparticles into commercial paints. In technology, titanium oxide (TiO2) is a crucial substance. As a paint pigment, it is widely utilized and exhibits improved performance in photocatalytic application for the removal of different organic pollutants from both air and water. TiO2 is also regarded as a promising option for photoelectrochemical energy production [12]. The semiconducting transition metal oxide material TiO2 has several advantageous characteristics, including easy controllability, low cost, non-toxicity, and robust resistance to chemical erosion. It can therefore be used in application such as environmental distillation, chemical sensors, and solar cells. The three most prevalent crystalline polymorphous forms of dioxide, which can also take on amorphous and crystalline forms, are anatase, rutile, and brookite. For titanium dioxide (TiO2), the aqueous medium is stable and can take both acidic and alkaline solutions. TiO2 nanoparticles have been used in photocatalysis, cosmetics and pharmaceuticals [13]. Naturally hydrophobic in most situations, TiO2 is a bright white, odorless powder. It is an opacifier that works quite well and is highly stable. TiO2 NPs, able to capture 3-4% of solar energy, are the most efficient solar collectors. One insoluble, fire-resistant, highly thermally stable, and non-hazardous metal oxide is titanium dioxide (TiO2). The atomic numbers of titanium (22 from the IV B group) and oxygen (8 from the VI A group) make up TiO2 [14].

II. Material and methods

A. Collection of Industrial Algae

Industrial algae from Siddhachalam Laboratory's (Raipur) stock was used in this investigation. The creation of TiO2  nanoparticles employed this algae as a template.

B. The formation of algae extract

Cleaned the Algae 5 to 6 times with normal water until the calcium is removed and The lastly cleaned the algae with distilled water. Dry until moisture is removed from the algae. 20g of algae powder are mixed with 150ml of D/W. and was kept in boiled under 2 hours. Then shake and centrifuge and filtrate.

C. Synthesis of TiO2 nanoparticles

About 50ml of distilled water was taken in a conical flask and 3.9933g of TiO2 was added to it. The mixture was varied the for magnetic stirrers 2 hours and their algal extract a resolution was added drop wise that an interval of 5minutes. The solution was washed (Centrifuged) by D/W. for 4 to 5 times in an interval of 20 minutes and was placed in centrifuge to obtain the precipitate and filtrate. After that the precipitate was washed with ethanol (Alcohol). Transferred to a glass beaker, is positioned on a Magnetic stirrer at low level (130 ͦ C) for drying. The dried precipitate was placed in crucible on Hot Air Oven under 110 ͦ C for 4 hours. After that the precipitate was transferred to Muffle Furnace for 4 hours at 500 ͦC and then it was cooled in a desiccator and collected. Then placed in a desiccators. After removing it from the desiccator, it is placed in another crucible and pistachios ground finely in the help of Mortar Pestle.

1. UV-Vis spectra analysis

TiO2 particles were described using a Shimadzu UV-Vis Spectroscopy 1900i. Since TiO2 nanoparticles are soluble in ethylene glycol, the sample must first be diluted with water or ethylene glycol before being placed in the UV-Vis Spectroscopy. This allows the sample to be directly in the light source, allowing the spectrum to be seen on the screen in the form of a graph. The absorption of the TiO2 nanoparticles was recorded in the transmittance mode in the region of 200–800 by UV-Vis light.

2. FT-IR

Using an FT-IR spectrum analyzer (Perkin Elmer's LS-65 Light-emitting spectrophotometer), the material composition of the synthetic titanium oxide nanoparticle was studied. The solution was described in the 4000-400 cm-1 range after being dried at a temperature of 75°C and powdered using the KBr pellet technique.

3. XRD

One special technique for figuring out a compound's crystallinity is XRD. A cathode ray tube is used to create the X-rays, which are then focused on the specimen, collimated to provide monochromatic radiation, and filtered. When conditions meet Bragg's law (nλ = 2dsinϴ), the interaction of the incident rays with the sample results in constructive interference (and a diffracted ray). Actually, one of the foundational ideas in the study of X-ray diffraction is Bragg's law. Bragg's law equation has an integer n, a characteristic wavelength (λ) representing the X-ray beams impinging on the crystallized sample, an interplanar spacing (d) representing the distance between rows of atoms, and an angle (ϴ) representing the X-ray beams relative to these planes.

4.Methylene Blue Degradation Analysis

Due to the solubility of TiO2 in water, the TiO2 sample solution with D/W was employed for MBD analysis. Several absorption graphs are created in each cycle of Vis Spectroscopy, which illustrates the metal oxide's degradation of Methylene Blue over a period of time.

5. For Antimicrobial study

TiO2 NPs are tested for antibacterial activity using the well diffusion method. The well diffusion method is frequently used to determine a compound's minimal inhibitory concentration as well as for antimicrobial testing of antibiotics and other metal compounds. This procedure involves dispersing the microbe to be tested, creating tiny wells via the borer, filling them with a metal oxide nanoparticle solution, and allowing the nanomaterials to diffuse. The impact of the NPs is to inhibit the growth of the organisms that are grown.

5.1 For Antibacterial Study

NAM media should be prepared for antibacterial study of TiO2 NPs

Peptone            - 5.00

HM Peptone B  - 1.50

Yeast Extract    - 1.50

Sodium Chloride           - 5.00

Agar                 - 15.00

D/W                 - 1000 ml

Final pH                       - 7.4 (0.2)

A pure culture of bacteria is inoculated by spreading and well-formed media that has been prepared and poured into Petri plates. TiO2 NPs solution is added in little amounts to the well, which are then examined and incubated at 37°C for the entire night.

5.2 For Antifungal Study

PDA media should be prepared for antifungal study of TiO2 NPs

Infusion from Potatoes   - 200g

Dextrose (Glucose)        - 20g

Agar                             - 15g

D/W                             - 1000ml

Final Ph                        - 7.4 (0.2)

After preparing the media, pouring it into Petri plates, and allowing it to solidify, a pure culture of a fungal colony is inoculated. A small amount of TiO2 NPs solution is introduced to the well, and it is then examined for 48 hours at 25°C during incubation.

III. Result also Discuss

v  UV-Visible spectra evaluation

The UV-Visual spectrophotometer serves as the foundation for the artificial materials UV-Vis spectrum NPs. The band gap of the produced TiO2 NPs was determined to be 3.4 by analyzing the UV-visible spectra, and we obtained the total absorbance for the NPs at that point. Based on UV absorbance spectra, we deduced that TiO2 reacts when UV light acts as a photo catalyst, breaking extraordinarily strong covalent bonds.Figure 1: Results showing UV-Vis spectraphotometer of TiO2 NPs synthesized from Industrial algae.

For band gap we use the Touc Relation, which is given by the formula;

hv = A (hv – Eg)n


hv = Incident Wavelength

Band gap calculation from the graph:

 = 3.4

Figure 2: Band gap of TiO2 NPs synthesized from Industrial algae.

v  FT-IR

The FT-IR spectrum used to examine the TiO2 NPs' functioning groups. To obtain an absorption frequency in the I region, an FT-IR spectrophotometer was employed. The stretching and bending vibration of the –OH groups is responsible for the spectra's peak at 3400 and 1650 cm-1. A minor peak at 1450 cm-1 is evident as the Ti-O-Ti stretching vibration. The functional groups that were present in the generated TiO2 NPs were identified using FT-IR spectra. Measurements were made between 4000-650 cm-1 at a resolution of 4 cm-1. The 800-1200 cm-1 range of the FT-IR spectra showed three prominent peaks. verifies the presence of many functional groups that suggest the extracts contain phenols, organic acids, and aliphatic amines. These substances may serve as reducing and stabilizing agents when the TiO2 NPs are being formed.

Figure 3: FT-IR spectrum of TiO2 NPs synthesized from Industrial algae.

v  XRD

Figure 4 shows the synthesized TiO2  NPs' X-ray diffraction pattern. The peak details are located at 36, 53, and 57, which correspond to the (004), (116), and (215) crystal planes, respectively.

Figure: 4 XRD spectrum of TiO2 NPs

v  Methylene Blue Degradation Analysis

Methylene blue is a chemical compound with the molecular formula C16H18C1N3S. Methylene blue was catalyzed to degrade by TiO2 NPs, and the extent of the degradation was assessed by UV-Visible spectroscopy. The visiblespectrum absorption peaks of methylene blue dye in water were found to be centered at 664 nm.

Figure 5: Showing dye for Methylene blue degradation.

The photocatalytic activity of TiO2 NPs during production was investigated by means of regular interval degradation of Methylene Blue Dye. For the test, 10-1 Methylene blue dye was utilized, and MB solution was significantly diluted with TiO2 NPs sample solution. The sample data is collected by UV-Vis Spectrophotometer for approximately 40 cycles at regular intervals of time, and the graph is produced. The modification of color MB in deep on light blue hue indicates a reduction in dye and photo catalytic effect. The photocatalytic activity of TiO2 NPs is demonstrated by the decrease in absorbance at various wavelengths over time.

Figure 6: TiO2 Photo catalytic Absorbance graph.

v  Antimicrobial Study

The solid agar medium is tested for microorganisms using the well diffusion method. Whereas no zone or no effect is observed in the fungal culture plate, indicating that no antifungal activity is observed in synthesized TiO2 NPs, an area of clear restriction around the well containing TiO2 NPs had been observed in the bacterial culture plate, demonstrating the antibacterial properties of synthesized TiO2  NPs.

             (A)                             (B)

Figure 7:  (A) TiO2 Antibacterial Study, Staphylobacillus

 (B)TiO2 Antifungal Study, Alternaria.


In order to cause a loss of barrier integrity, an increase in membrane fluidity, cellular content leakage, and ultimately cell lysis, TiO2 NPs have been focused on the oxidation of phospholipids and its impact on membrane integrity.


Finally, we can say that using a green synthesis approach, the process of creating nanoparticles was effectively finished. The FT-IR data confirm the vibration NPs of TiO2, and the UV-Vis spectroscopy absorbance data conformance regarding the absorbance and the result reveals that the NPs were manufactured appropriately. The absorbance can be observed at 320 nm, which is similar to the absorbance of TiO2 NPs and allows us to compute the band gap of around 3.4. The Methylene Blue Reduction Test verifies the photocatalytic effect, and the dye reduction of Methylene Blue over around 40 cycles over the course of an hour is demonstrated by the blue color becoming lighter and a removal percentage of roughly 70%. By preventing the bacteria Staphylobacillus from growing, antibacterial activity demonstrates the antimicrobial properties of NPs.

The clear zones surrounding the well holding the TiO2 NPs are established after the incubation period of one day. We can also conclude that TiO2 NPs are a convenient antimicrobial agent for use against bacteria.

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