Characterization of Bacterial Cellulose (BC)
Bacterial cellulose is a natural occurring organic compound. It has the chemical symbol of C6H10O5. According to Agarwl, Palph, Reiner and Verrill (2017), state that bacterial cellulose can be characterized by its structural components. The structural, component of a bacterial cellulose vary depending on the purity, strength of the bonds and the porosity. The purer the bacterial cellulose, the stronger and less porous it is (Agarwal et al., 2017). It is important to characterize bacterial cellulose since different bacteria can be applied in different fields such as biotechnology, microbiology, and materials science (An et al., 2017). In fact, proper identification of the cellular structure of the bacterial cellulose is important in enhancing the flavor and color of processed foods.
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An example of such a characterization is the bacterial cellulose used in the manufacture of Nata de Coco (Aslan & Anik, 2016). Moreover, proper characterization of the bacteria leads to the efficient synthesis into cosmetic and even soft textiles. Characterization enhances product specialization and it can be analyzed using different methodologies, for instance, Fourier Transform Infrared Spectroscopy (FTIR), x-ray diffraction, Field Emission Scanning Electron Microscope (FSEM), and thermogravimetric analysis.
Fourier Transform Infrared Spectroscopy (FTIR)
Fourier transform infrared spectroscopy (FTIR) is a method use to pass infrared spectrum drawn from the synthesis of a solid, liquid or gas. According to Wang, Huang, Y LI and Liang (2016), using Fourier transform infrared spectroscopy, researchers can determine the cellular composition of things. FTIR is most preferred in determining the inter- and intra- molecular hydrogen bonds in cellulose (Wang et al., 2016). Bandyopadhyay, Smarak and Petr (2018), the Fourier transform infrared spectroscopy uses photolysis under different cryogenic temperature variations, to test the specific protein–ligand bonds between the structure of solids, liquids and gases. Fourier transform infrared spectroscopy (FTIR) is also used in Raman Scattering. Rama scattering refers to the technique used evaluate the strength emitted during the dispersion of particles in their different states.
The relationship of Fourier transforms infrared spectroscopy (FTIR) and bacterial cellulose is that the method is used to analyze the characterization using a Hestrin-Schramm medium. In addition, Torres, CCrohua and Troncoso (2019), state that the use of Fourier transforms infrared spectroscopy (FTIR) is fast, reliable, inexpensive, and time efficient. The Fourier transforms infrared spectroscopy (FTIR) has a transmittance mode of a wavelength ranging from 4000 to 400 cm-1. Moreover, the Fourier transforms infrared spectroscopy (FTIR) captures information relating to both the physical and chemical characterization of the bacterial cellulose under investigation. The preference of the Fourier transforms infrared spectroscopy (FTIR) in the characterization of bacteria Gluconobacter is the ability to initially freeze and convert the material into powder. The Fourier transforms infrared spectroscopy (FTIR) has a detector that monitors the wavelength range and transmits the signal to a computer which translates the signal into an absorption spectrum.
FTIR can be used in the characterization of various forms of Gluconobacter oxydans including the strain ATCC 621H strain ATCC 621H. According to Markov, Frece, Pleadin and Bevardi (2019), the cellular components including the proteins, membrane fatty acid and oligo- and polysaccharides had various functional groups varying in their IR-absorption frequencies. The specifications across the different cellular structures leads to significant alterations in corresponding IR spectra. Markov et al., (2019), and Bandyopadhyay et al., (2018), agree on the premise that the variations in the cell surface of viable and heat-treated cells of G. oxydans leads to the dominant absorption of bands (See table below).
Kim et al., (2016) use the sorbitol dehydrogenase (GoSLDH) from Gluconobacter oxydans G624 (G. oxydans G624) as expressed in Escherichia coli. According to their findings, the presence of additional observed peaks at 1,600–1,800 cm−1 in the Fourier transforms infrared spectroscopy spectra of GoSLDH on particles, has a strong positive correlation to the amide bonds and -N= C=O and C=C stretches. Shaban, Mohamed and Abdallah (2018) take on a different approach with their use of the ZnO precursor concentration. Yusof, Dipak, Ahmad and Takeshi (2016) agree with the aforementioned premise. The results of their study show that FTIR spectra of two samples coated at pH 6 and pH 8 have stronger vibrational modes, Moreover, as the percentage of the ZnO increases, the percentage of the OH% increases too. The final result is the formation of a thick layer of zinc oxide on the gluconobacter. Wang, Liang, Huang and Li (2016) use FTIR on Gluconobacter in their analysis of techniques to catalytically convert carbohydrates into lactic acid (LA). The results of the aforementioned study confirm that the low temperature desorption indicate the presence of weak acidic sites in the material. The result is similar to that of Tabarsa, Sheykhnazari, Ashori, Maskour and Khazaeian (2017), in their analysis of nano bacterial cellulose. Yusof, Rana, Ismail and Matsuura (13) use the FTIR on Gluconobacter to analyze the microstructure of the polyacrylonitrile among various carbon fibers. FTIR spectroscopy was used to observe the presence of functional groups and chemical structure transformation of PAN/AM fibers prepared under various heat treatment stages of the fiber. The result of their study shows that porosity increases together with the loss of volatile components through the activation process (Yusof et al., 2016: Chen et al., 2016). The importance of the study by Yusof et al., (2016) is that it shows the varied manifestations of physical activation of carbonized AM/PAN fiber using carbon dioxide gas.
X-ray Diffraction
X-Ray Diffraction is hapless procedure that offers specific details of the crystallographic anatomy, chemical constitution, and physical structure of materials. According to Torres, Ccorahua, Arroyo and Trancoso (2019), X-ray diffraction shows the crystalline pattern of unmodified cellulose I up to moderate degrees of acetylation. The variation in peak widths shows that acetylation proceeded from the surface of micro fibrils, leaving the core portion unreacted. Agarwal et al., (2017) use X-ray diffraction to study the patterns in moisture-content using the bacterial cellulose crystallinity and the variable sizes. The findings of the study affirm that using the various cellulose samples, the effect of moisture change upon diffract gram derived parameters was investigated. Karimi and Taherzadeh (2016), state that non-structural carbohydrates, extractable ash, proteins, starch, and high extractive contents, re-localization and cell wall delamination. On the other hand, Auta, Adamus, Kwiecien, Radecka and Hooley (2017), use x-ray diffraction to investigate the production and subsequent characterization of the bacterial cellulose prior to the enzymatic hydrolysis. The findings by Auta et al., (2017) affirm that X-Ray Diffraction (XRD) revealed a high purity of BC indicating type I cellulose with high crystalline nature. Fan, Dai and Huang (2012) use X-ray diffraction to analyze the X-ray diffraction has been a powerful tool to investigate hydrogen bonds, visualization, lengths and angles in natural fibers. Fan et al., (2012) and Shao et al., (2016), affirm that the molecular orientation, crystallization and formation of micro fibrils differ from one plant to another as well as different environments and other physical effects.
According to Feng et al., (2015), the physio-chemical and morphological structure of the bacterial cellulose produced in Super Optimal Broth are determined using powder X-ray diffraction (pXRD) among other tools. The study shows that the use of Super Optimal Broth and X-ray diffraction enhances the yield of bacterial cellulose and allows conversion of 50% of the carbon source to bacterial cellulose. The yield is much higher as compared to a marginally low 7% conversion in the case of traditional Hestrin–Schramm medium after 7 days of interaction. The importance of the study by Feng et al., (2015), lies in the application bacterial cellulose in food industries, that still require exploration. Specialization is important in such a volatile industry as companies seek to reduce their work force, time, costs, waste, and environmental pollution.
Field Emission Scanning Electron Microscope (FSEM)
Field Emission Scanning Electron Microscope (FSEM) visualizes particularly minute topographic details on the surface or entire or fractioned objects. Sajjad et al., (2019), state that the FSEM works with electrons using particles with a negative charge as opposed to other visualizations that make use of light. Sakeena, Sven, Bernhard, Philip, Wolfgang and Falk (2018), agree with the aforementioned assessment as applied in their research of carbohydrate polymers. The electrons in the FSEM tool are liberated by a field emission source. In retrospect, the object under observation in the FSEM is scanned by electrons according to a zig-zag pattern. An et al., (2017), uses the FSEM to understand the reaction of cellulose membranes treated under the Electron Beam Irradiation as used in guided bone regeneration. Santos et al., (2015), analyzes the Bacterial cellulose from the Nata de coco product, under the high visualization offered using the FSEM, has high purity and properties presents a potential biomaterial in various industries. Lotfiman, Biak, Tey, Suryani and Saeid (2018), through the findings of their studies agree that the high cellulose is used in industries such as the biomedical/cosmetics, electronic/smart materials applications and reinforcement agent in composites. Santos et al., (2015), highlight the morphological changes, as projected from the alkaline environment and ultrasound specifications on a commercially available bacterial cellulose such as that sampled from Nata de coco.
Haghighi, Abbaspour, Karimi, and Fattahi (2016), use the high imaging capacity of FSEM for the eco-friendly synthesis and antimicrobial events surrounding their case sample of Silver Nanoparticles. The partciles used by Haghigh et al., (2016) use the Dracocephalum moldavica Seed extract that contains the gluconobacter cellulose including Escherichia coli, as a significant part of the membrane. The importance of the use of the highly visualized FSEM tool in the analysis by Haghigh et al., (2016) is the synthesis of nanoparticles (NPs) has over time evolved into a matter of significant interest due to their varying return properties and applications in different fields. Currently, various strategies have been proposed in light of the generation of metallic nannoparticles including electro-chemical and other photochemical processes. On the other hand, a considerable number of the techniques suffer from the use of invariably high toxic, hazardous chemicals that result in significant challenges in purification.
Thermogravimetric Analysis
Thermogravimetric analysis is the deep outlook of the weight changes as a function of temperature and time of particles. According to the introductory definition of thermogravimetric analysis by Liu, Wang, Cheng, Sheng and Yang (2019), understanding the weight changes is crucial in understanding the variations of polymeric materials as a result of decomposition and oxidation reactions. Other changes in the weight physical processes include sublimation, vaporization, and desorption of particles (Frone et al.,2018). Pacheco et al., (2017) adds unto the discussion by stating that the importance of the use of thermogravimetric analysis in the analysis of Gluconobacter and their molecular structure as polymers and their transition provides information regarding changes in mass of the materials due to change in temperature or time in a controlled manner. Aslan and Anik (2016), uses thermogravimetric analysis to develop microbial glucose biosensor to understand the adaptation of this favorable system to G.oxydans based microbial biofuel cell.
Thermogravimetric analysis of bacteria ensures the synthesis of inorganic frameworks with specific and organized core networks is of potential advantage in processes such as catalysis, separation technology and biomaterials engineering. According to Shao et al., (2016), thermal processing, also known as thermogravimetric analysis, is the most preferred technology applied in the preservation of food materials. The main goal of thermal processing is to inactivate the spoilage and pathogenic microorganisms and produce a safe product with enhanced shelf life. The studies by Pa’e et al., (2019), to understand the effect of microbial inactivation under heat is potentially important in maximizing options for heat treatments to reduce and eradicate foodborne disease. In this light, the risk of spinning that is a result of common and emerging strains while circumventing over the manufacturing of the food material.
Thermogravimetric analysis has led to the realization that microorganism’s degeneration is linked to the irreversible denaturation of membranes, ribosomes, and nucleic acids. On the contrary, the trends of macromolecular changes that foster the cell death of microorganisms during thermal treatment is still vague. Karimi and Taherzadeh (2016), agree with the aforementioned trends of macromolecular changes with reference to using differential scanning calorimetry. Under a specific thermal temperature, the bacteria strain can transition from one thermophile to another (Kumar et al., 2019: Semjonovs et al., 2016). In conclusion, the aforementioned studies show a significant correlation in the calculated viability drawn from the calorimetric data. Moreover, and calorimetric data drawn from thermogravimetric analysis offers a different set of data unique information using the direct measurement of the thermodynamic energy necessary to inactivate microorganisms.
To conclude, evaluating and quantifying variances among the thermograms of entire cells that have specific components permits a researcher to rank the varying ranges of thermal stabilities of the various cellular structures and identify those most vulnerable to thermal disruption. Kumar et al., (2019) state that Polymers reinforced with nano fillers offer promisingotions foraconiderable number of applications. The definition of the exact weight of a polymer filter is drawn from analyzing its weight using thermogravimetric analysis. Moreover, Faria et al., (2019) states that the preferred characteristics is lightweight and high-performance, multifunctional materials. Du et al., (2018) focuses his study on various techniques of visualization including thermogravimetric analysis. According to Du et al., (2018), the fundamental characteristics of Nano sized materials that are responsible for their specific characteristics are the nanometer length scales (available in thermogravimetric analysis) and significantly large surface area-to-volume ratio. Using the thermogravimetric analysis the researchers can come up with new applications in aqueous systems, including replacing chemicals and dyes with water while retaining the native form and properties of the pellicle. In this regard, more bacterial cellulose is used in making natural hydrogel material.
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