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Preparation and application of nanoparticles and nanofibres with natural UV filters
Plachá, Monika ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
The presented diploma thesis is focused on preparation of nanoparticles and nanofibres with natural UV filters. Liposomes with encapsulated aqueous, ethanol and lipid extracts were prepared. Nanofibers from PHB containing lipid extract were prepared too. As a part of this work, an overview of natural sources with potential effects as UV filters were introduced. Moreover, nanoparticles and nanofibers and methods of their characterization were described. Size, polydisperse index and colloid stability of prepared nanoparticles were characterized via DLS. In experimental part aqueous, ethanol and lipid extracts were prepared from roasted coffee, green coffee and cascara. These extracts were spectrophotometrically characterized for the content of polyphenols, flavonoids, antioxidant activity, tannins and their SPF. Liposomes and liposomes containing PHB with these extracts were prepared and the encapsulation effectivity, short–term and long–term stability as well as SPF of nanoparticles were determined. Nanofibers from PHB containing lipid extracts were prepared via electrospinning and forcespinning methods. Prepared nanofibers were examined via FTIR–ATR. Antioxidant activity, short–term and long–term stability were determined spectrophotometrically. From selected nanoparticles, emulsions and gels were prepared and their SPF was also determined. Three types of emulsions with the best SPF were selected and tested on volunteers. Sedimentation stability of emulsions was tested by analytical centrifuge. Finally, cytotoxicity of selected nanoparticles and nanofibers was tested via MTT assay using human keratinocytes.
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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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Study of selected antibiotics sorption on polyhydroxybutyrate
Amrichová, Anna ; Doležalová Weissmannová, Helena (referee) ; Řezáčová, Veronika (advisor)
The bachelor thesis deals with the issue of sorption of antibiotics on microplastics. Antibiotics and microplastics are now a highly studied topic, due to the positive and negative properties. Antibiotics with groups of sulfonamides and quinolones were selected for the experiment. From sulfonamides, sulfamethoxazole and trimethoprim were selected, and from the quinolones, enrofloxacin and ciprofloxacin, which have already been the impetus for some microplastic studies. Biodegradable polyhydroxybutyrate was used as the microplastic. Two experiments were performed in the laboratory. The first aimed to monitor the time dependence of antibiotic sorption on polyhydroxybutyrate, the second the concentration. The concentration dependence was subsequently evaluated using the Freundlich and Langmuir isotherms. The method of liquid chromatography with mass spectrometry was used for analyzes.
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Crystallization of microbial polyesters - methods and biological consequences
Liczka, Jan ; Kalina, Michal (referee) ; Sedláček, Petr (advisor)
The aim of this bachelor thesis was to test suitable methods for preparation of amorphous form of microbial polyesters and to perform model experiment to study a kinetics of crystallization of prepared polyesters. The theoretical part deals with PHA characteristics and various methods of isolating PHB granules from bacterial cells. In the experimental part, measuring techniques such as analytical centrifugation, dynamic light scattering (DLS) and Fourier transform infrared spectroscopy (FTIR) were tested to characterize the properties of PHB granules. Furthermore, an experiment was performed to isolate native PHB granules from Cupriavidus necator using lysozyme, deoxyribonuclease and cell disruption by ultrasonification. The results of the analyzes showed that the granules were either isolated amorphous only in the wet and crystallized upon drying, or that the granules were completely amorphous but not isolated from the bacterial cell.
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Biodegradation of polyesters and other polymers in soil and compost
Papala, František ; Smilek, Jiří (referee) ; Obruča, Stanislav (advisor)
This diploma thesis studies the biodegradation of polyhydroxybutyrate (PHB) and polylactic acid (PLA) within soil and compost environments. The experimental part of this work is concerned with the differences in degradation across environments and the effects of degraded polymers on the environment itself. Experiments revealed that higher temperature and potentially also higher humidity substantially affect the rate of biodegradation. A significant degradation of all the polymers was observed in the compost medium, while polymers within the soil medium did not demonstrate any visible change nor change in mass after the 8th week of experiment. Even though SEM images revealed slow microbial degradation of soil PHB, this was not seen in any of the soil-incubated PLA samples. This might indicate primarily abiotic mechanism of PLA degradation. Germination and grow tests, which used the prepared polymer-enriched compost as a substrate and fertilizer, respectively, did not in most cases reveal any significant differences between individual plants across substrates. This seems to imply that neither the examined polymers nor their degradation products generated during composting are phytotoxic.
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Hybrid biopolymer composites for 3D printing applications
Menčík, Přemysl ; Bakoš, Dušan (referee) ; Dzik, Petr (referee) ; Jančář, Josef (advisor)
This dissertation work deals with the thermic and the mechanical behavior of plasticized bio-plastics and bio-composites for the 3D printing applications. The influence of plasticizer chemical structure on thermic and mechanical properties of plasticized polymeric blends from the poly-3-hydroxybutyrate and the poly lactic acid was investigated. Used plasticizers are based on derivative of citric acid. The influence of plasticizers on polymeric matrix and their compatibility was estimated by gear torque rate of melt mixer, respectively rate of plasticizer migration from the material during higher temperature. The plasticizer structure influence on the glass transition temperature and on the kinetics of crystallization of plasticized material was investigated by modulated differential scanning calorimetry. The behavior of material during 3D printing was also observed. Mechanical properties of printed samples, especially their elongation at break, were determined by tensile tests. The largest softening effect was observed using tributylcitrate plasticizer, where the glass temperature decreased by 35 °C and elongation at break increased by 150% compared to non-plasticized reference material. This plasticized polymeric blend showed also sufficient 3D printing properties and was used as the matrix for composites in the next part of this work. Composites were filled by kaolin, limestone, halloysit, fumed silica, talc, magnesium hydroxide and chopped flax fibers. Particle distribution in composites in dependence of used surface treatment of filler was observed by scanning electron microscopy. The influence of composite filler on rheological properties, crystallization kinetics and thermal stability of composites, was observed by viscometry and differential scanning calorimetry. Their mechanical properties and heat deflection temperature were observed on samples prepared by 3D print. Kaolin in composite material showed homogeneous particle distribution and insignificant nucleation effect and influence on thermic stability. Composite filled by kaolin also showed 18% smaller warping during 3D printing compared to non-filled reference. Consequently kaolin was evaluated as suitable inorganic filler for bioplastic composite intended for 3D print and this composite was used in the following part of this thesis. Method of mathematical prediction of Young's modulus was described for composite samples prepared by 3D print. Composites filled by one type of filler – kaolin, or limestone, resp. by combination of both fillers were investigated on the basis of the micromechanic Halpin-Tsai model modified by the semiempiric multiparametric Cerny's equation. Additive and combinational method of Young's modulus evaluation is used for composites with hybrid filling. Deflection of measured and theoretical Young's modulus value of composite filled with kaolin was decreased by established correction from 21% to 1% and for composites filled with limestone from 13% to 9%. In this manner it is possible to predict the Young's modulus of the samples prepared by 3D print.
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Ecotoxicological evaluation of selected plastics
Demová, Kristína ; Mravcová, Ludmila (referee) ; Zlámalová Gargošová, Helena (advisor)
The presented bachelor thesis deals with the issue of plastics in the environment and their ecotoxicological evaluation. This topic is currently being brought to the fore due to the development of new, more easily degradable plastics. This is intended to be a solution to the current environmental problem. That is why the aim of this work was to verify the assumption of ecological safety of such plastics. The experimental part focuses on one selected biodegradable plastic - polyhydroxybutyrate, which was evaluated using acute and chronic toxicity tests. The tests were performed in an aquatic arrangement to predict the effect on the aquatic ecosystem. Two organisms were chosen from its representatives, the crustacean Daphnia magna and the plant Lemna minor. The chronic test on the organism Daphnia magna shows the greatest results, which indicates the negative effects of the selected plastic on the tested organism.
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Photoprotective properties of PHB granules
Hesko, Ondrej ; Obruča, Stanislav (referee) ; Mravec, Filip (advisor)
The objective of this work is study and design of physical-chemical methods for verification of photoprotective properties PHB inclusions using bacteria Cupriavidus necator H16 and its mutated bacteria strain Cupriavidus necator PHB-4 which differs in production capability of polyhydroxyalkanoates. Basic information about of polymer inslusions and methods used for proving photoprotectivity are discussed in theoretical part. Methods such as nephelometry and continous fluorescence microphotolysis (CMF), which is used to characterize the mechanism of interaction of light with inclusions, were optimized. In experimental part properties of light scattering in bacteria were determined and characteristic curves were obtained. Further the photostability of fluorescence probe specific for nucleic acids were compared in bacteria strains. This method should prove bigger tolerance of bacteria H16 for UV light. It should also define possible interaction of nucleic acids with PHB inclusions.
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Isolation of PHA from bacterial biomass and their subsequent purification
Zlochová, Barbora ; Kouřilová, Xenie (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis deals with the isolation and characterization of the bacterial polymer PHA isolated from the bacterial strain Halomonas halophila (CCM 3662) by osmotic stress in distilled water, ie hypo shock, when the cells crack and release PHA. The addition of biosurfactants simultaneously solubilizes the proteins and lipids of the residual biomass, theoretically achieving a purer PHB product without the use of toxic substances such as chloroform. The theoretical part of the work focuses on PHA in terms of their general properties and use, the mechanism of synthesis of PHA microorganisms and their subsequent degradation and the existing strategy for isolating PHA from bacterial biomass. The experimental part describes the production of PHB bacteria Halomonas halophila and the subsequent process of isolation of PHB by hypo shock of halophilic bacteria in distilled water with the addition of selected biosurfactants. Biosurfactants used in the work are coconut soap, lecithin, rhamnolipids, sophorolipids, coco glucoside, lauryl glucoside and cocoamidopropyl betaine. The properties of the isolated PHB material were analyzed by gas chromatography with FID detection and SEC-MALS and FTIR methods. Based on the results, we chose the pair of coco glucoside and lauryl glucoside as the most suitable biosurfactants that can be used to isolate PHB from bacterial biomass. In combination with washing the material in ethanol, at concentration of biosurfactant 5 g/l and isolation in 70 ° C, it was possible to achieve a purity of 90 % of material using cocot glucoside and 89 % purity of material using lauryl glucoside.
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Detoxification of lignocellulose hydrolyzates
Vašíčková, Monika ; Pořízka, Jaromír (referee) ; Obruča, Stanislav (advisor)
The aim of this work was study of the detoxification of lignocellulose material hydrolysates and to investigate sawdust suitability as a substrate for microbial production of PHA by bacteria Burkholderia cepacia and Burkholderia sacchari. In the experimental part of the work the most suitable way of detoxification of model and real hydrolysate was studied. After that, detoxification methods used were evaluated. Criteria for evaluation were concentration of polyphenols as the most important microbial inhibitors and reduction saccharides as the main carbon substrate. Furthermore, fermentability of the hydrolysates was also tested by cultivation of two bacteria capable of PHA accumulation. Burkholderia sacchari demonstrated higher ability to accumulate PHA then Burkholderia cepacia. Then in the summary – most effective way for detoxification was ‚overliming‘. Major increase of PHB in biomass was obtained when Burkholderia sacchari was cultivated on media gained by application of overliming of real lignocellulose hydrolysate. However, total gains of PHB were more likely low and then sawdust can not be considered as a substrate for PHB production at industrial scale.
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