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Raman spectroscopy as a tool for analysis of microbial cells
Pokorný, Petr ; Enev, Vojtěch (referee) ; Samek, Ota (advisor)
This bachelor thesis is focused on the use of Raman spectroscopy for analysis of ectoine and hydroxyectoine presence in bacterial strain Halomonas elongata. Theoretical part compile characteristics of extremophiles with closer look on halophilic organisms and one of their main osmolytes, ectoines. Following by description of Raman spectroscopy method and its uses. Experimental part deals with cultivation of bacteria on different combinations of substrate and salt concentration in order to reach optimal production of PHA and for achieving the highest possible yield of biomass to be analysed by Raman spektroskopy. As the best substrate for PHA production turned out to be glukose along with salt concentration 30 g/l NaCl, where percentage representation of PHB makes 30,5229 %. As an ideal substrate for the highest yield of biomass proved to be sacharose. Bacteria that grew on sacharose with three different salt concentrations, 40, 70 and 100 g/l NaCl was measured along with Halomonas salina and Halomonas organivorans for comparison. Raman spectroscopy unfortunately turned out to be inadequate for measurement of this type of sample, thus we were not able to measure ectoine of hydroxyectoine presence in cells.
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Analytical methods for determination of polyhydroxyalcanoates content in bacterial cells
Černayová, Diana ; Enev, Vojtěch (referee) ; Sedláček, Petr (advisor)
This bachelor thesis deals with methods that are used for determination of polyhydroxyalkanoates (PHA) in cells of bacteria Cupriavidus necator H16. To the applied methods belong Fourier transform infrared spectroscopy, Raman spectoscopy, turbidimetry and thermal analysis (thermogravimetric analysis and differential scanning calorimetry). The results of each method were compared to amount of PHA determinated by gas chromatography. Fourier transform infrared spectroscopy and Raman spectroscopy revealed a linear relation between ratio of characteristic peaks and amount of PHA. FTIR enabled to determine not only the quantitative amount of PHA, but the quality of the sample (cristallinity, amorphous state) as well. Another method was turbidimetry, which has shown correlation between dispersion of UV-VIS light to amount of PHA in bacterial cells. Thermogravimetric analysis was the most effective technique for determination of PHA. The dependence of weight loss of PHA on its content (found by gas chromatography) was determined with the highest accuracy to gas chromatography. Tha last method- DSC was not alternative replacement for quantitative PHA determination. However, DSC indicated crystalline and amorphous state of the polymer.
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Stability of soil organic matter and humic substances
Nováková, Šárka ; Enev, Vojtěch (referee) ; Klučáková, Martina (advisor)
This diploma thesis is focused on changes of stability in organic matter by extraction in different agents. Two soils of a different type and isolated humic acids were used for stability determination. Extraction agents were selected usually used for soil metal extraction, and a changes in the structure of the organic matter was observed. Samples were characterized using FTIR analysis and elemental analysis, the extracts were measured by UV-VIS spectroscopy, absorption ratios E2/E3, E4/E6 were discovered, dynamic light scattering were determined for particle size distributions. Next part of the thesis was the assessment of the change of thermal stability using thermogravimetric analysis and differential scanning calorimetry, degradation temperatures of the extracted samples were discovered and compared with the original samples.
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Characterization of humic substances isolated from natural waste treated by vermicomposting process
Marková, Kamila ; Doskočil, Leoš (referee) ; Enev, Vojtěch (advisor)
This bachelor thesis deals with the study of humic substances isolated from vermicompost natural waste through spectrometric methods. The aim of this thesis is to study physicochemical properties of the humic substances through modern instrumental techniques such as molecular absorption spectroscopy in the ultraviolet and visible regions of electromagnetic radiation (UV/Vis), infrared spectroscopy with Fourier transformationn (FTIR), elemental analysis (EA), thermogravimetric analysis (TGA) and inductively coupled plasma emission (ICP OES). A research focused on basic information about the humic substances was developed in the theoretical part. A description about their distribution or structure was made and also their utilization in medicine, industry or ecology. Then, methods, which are used for study of the humic substances, were described as well. An essential part of the thesis was isolation of humic and fulvic acid from the vermicompost natural waste. Absorption coefficients were calculated from the measured UV/VIS spectrum which served to following characterization of HK and FK. FTIR spectrum was used to determination of functional groups of studied HL. An area of biogenic elements was specified using the elementary analysis. Total amount of studied trace elements was determined with with the help of ICP-OES.
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Experimental study on the release of active ingredients from gel matrices
Volhinava, Anastasiya ; Enev, Vojtěch (referee) ; Sedláček, Petr (advisor)
This thesis deals with the characterization of hydrogels containing humic substances and salicylic acid. Experimental study of the transport of active substances was performed from the gel matrix over the skin and synthetic membranes. During this work was writing literature review focused on the use of gels with humic substances in cosmetic, medical and pharmaceutical industries. The model humic hydrogels was prepared with salicylic acid and their characterization by basic methods of material analysis. These basic methods were and visual assessment of consistency during the preparation and rheology. The study of the release of active and humic substances was performed due to the vertical diffusion cells. The amount of released humic substance was characterized by UV-VIS method and the amount of released salicylic acid from gel matrix was determined by HPLC.
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Decomposition and stability of organic matter in soil
Tabaková, Eva ; Enev, Vojtěch (referee) ; Klučáková, Martina (advisor)
In the soil, the plant material is decomposed by several factors. In the process, one part of the carbon returns to the atmosphere as carbon dioxide, whereas another part is stabilised in the soil. The aim of this study is decomposition and stability of organic matter in soil using a simple and innovative TBI method. Through this method, we examined the weight change after the incubation period of 90 days, where biomass of green tea was degraded in cambodia soil. Tea bags have provided us with valuable information on the decomposition and amount of carbon in the soil, because the biomass itself is of organic origin. This information is based on the fact that the soil and the decomposition processes have a direct impact on the change in climatic conditions. Thanks to further analytical methods, elemental analysis, thermogravimetric analysis and infrared spectroscopy with Furier transformation, was found indispensable information about tea composition.
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Raman spectroscopy as a tool for analysis of biotechnologically relevant microorganisms
Záhorská, Linda ; Enev, Vojtěch (referee) ; Mgr.Ota Samek, Ph.D. (advisor)
The diploma thesis deals with the study of biotechnologically significant microorganisms, using the Raman spectroscopy. Content of the theoretical part is brief characteristic of Raman spectroscopy as a method, its use in practice and also use as a tool for monitoring of biotechnologically processes. Thesis was further focus on the biotechnologically significant microorganism Aureobasidium pullulans, its use in biotechnology and also for over-produced substances and in particular poly-L-maleic acid and pullulan. The content of the experimental part was study of selected strains A. pullulans, specifically stains as DSMZ, CCM F148 and CCM 8182, using Raman spectroscopy on the various types of culture media. Subject of practical part research was too production of extracellular polymers, acid poly-L-apple and pullulan, by selected strains A. pullulans. Objective of my thesis was described and determinate, spectra of individual strains as well as extracellular products, mainly pullulan, and then choose suitable production medium and optimal production strain A. pullulans. During experimental work was found, that optimal production strain was DSMZ strain culture on the mineral medium with the addition of yeast autolysate, which was optimal medium type. The content of the pullulan produced was for gravimetric determination, 6,3g/L, which also confirmed the results of the HPLC method. It was experimentally found, that Raman spectroscopy isn´t suitable method for quantification of extracellular products, but is appropriate and was used for PCA analysis of individual strains.
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Study of biological activity of superabsorption polymers
Männlová, Adriana ; Enev, Vojtěch (referee) ; Klučáková, Martina (advisor)
This thesis is focused on the study of the biological activity of superabsorbent polymers for environmental and agricultural applications. Generally these hydrogels perform many functions in the soil, which is increasingly stressed by treatment with synthetic fertilizers and thus loses their natural properties, such as the uptake of moisture. Addressing these issues is now in superabsorbent polymers that can absorb and also retain a lot of water around the root system as long as possible. They can perform the function of carriers gradual release fertilizer. Based on the literature review was designed and conducted the study of biological activity in the soil model on corn sown. SAPs were prepared in the framework of contract research Faculty of Chemistry of the Technical University in Brno with the company Amagro Ltd. The content of the experiment, the superabsorbent polymer, observing effects on the growth of dent corn. He was also studied root system for absorption of nutrients. Further changes were observed rheological properties of prepared superabsorbent polymer, superabsorbent polymers since they provide a source of nutrients and help to retain moisture in the soil, it is necessary to know their viskoelstick properties even at temperatures below freezing. This work is based primarily on the use of great motivation superabsorbent polymers in agriculture and also in environmental protection.
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Carbon nanoparticles from lignite
Kohoutková, Eliška ; Enev, Vojtěch (referee) ; Pekař, Miloslav (advisor)
These bachelor thesis focuses on lignite as a potential raw material for the carbon quantum dots extraction. There was proposed mechanochemical process of isolation these nanoparticles from lignite based on literature search. Lignite was mechanically stressed and subsequently oxidized to hydrogen peroxide. The results show that proposed method will have to be modified but lignite could be used as a feedstock for the carbon quantum dots extraction.
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Fractionation and molecular organization of humic acids
Chytilová, Aneta ; Enev, Vojtěch (referee) ; Klučáková, Martina (advisor)
Humic acids are part of the natural organic matter occurring all around us. The aim of this thesis is to study the molecular organization, conformation of humic acids in aqueous solutions, that always raise a number of questions. For a long time, the scientists all over the world argue, if humic acid are polymers, micelles or supramolecules. Over time, thanks to new technologies, their opinions are moving away from a polymer model and tend rather to supramolecular arrangement of humic acids. Studying humic complex systems is not easy, because they are polydisperse and heterogeneous, which significantly complicates any characterization. Moreover, its molecular organization is affected by many factors such as e.g. pH, ionic strength and etc. For the study of the conformation of humic acids, concentration series of IHSS (International humic substances asociation) Leonardite humic acids stamdards in four different mediums were prepared: 0,1 M NaOH, humic acids in water with pH modified to 12 (basic medium), 0,1 M NaOH + 0,1 M HCl, 0,1 M NaCl (neutral environment). Furthermore it has been performed the fractionation of humic acids for the purpose of simplifying the complicated structure. Prepared concentration series were characterized with several analytical methods such as ultraviolet and visible spectroscopy, dynamic light scattering, electrophoretic light scattering, microrheology, gel permeation chromatography, potentiometric pH determination and direct conductometry. Diploma thesis is built on the previous bachelor thesis in which different sample of humic acids was studied. The measured results indicate that the studied systems show supramolecular behavior and in some cases are subject to aggregation into larger units (micelles).
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