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Polyhydroxyalkanoates and their role in bacterial biofilms
Rucká, Markéta ; Samek,, Ota (referee) ; Obruča, Stanislav (advisor)
This master thesis deals with polyhydroxyalkanoates (PHA) and their role in bacterial biofilms. In the theoretical part the polyhydroxyalkanoates, bacterial biofilm and the relationship between them were reviewed. The experimental part focused on differences in PHA production by planktonic and biofilm cells. In order to study selected topic, bacterial strains of Burkholderia cepacia and Burkholderia sacchari were cultivated using a CDC biofilm reactor. The attention was paid to quantity and especially to the form in which PHA occurs in planktonic and biofilm cells. Results of Raman spectroscopy have shown that PHA exists exclusively in native amorphous form in planktonic bacterial cells. On the other hand, in biofilm PHA occurs also in a partially crystalline form. In addition, the resistance of planktonic and biofilm cells against various stress factors and the effect of osmotic stress on PHA production was tested too. According to the results of the experiment, when the bacteria were exposed to different stress factors (high temperature, low temperature, presence of detergent and so forth) biofilm cells showed a higher stress resistance than planktonic cells. Apart from slowing cell growth and reproduction, increased osmotic pressure in the culture medium also caused decrease of PHA production. In addition, planktonic cells responded to external stimuli more sensitively than biofilm ones.
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Modification of polyurethane by biodegradable poly(hydroxybutyrate)
Kupka, Vojtěch ; Žídek, Jan (referee) ; Vojtová, Lucy (advisor)
Cílem předložené bakalářské práce byla syntéza elastomerního polyuretanu plněného biodegradabilním polyhydroxybutyrátem. Vzorky byly podrobeny analýze morfologie pomocí rastrovací elektronové mikroskopie, mechanické vlastnosti byly testovány jednoosou tahovou zkouškou. Práce se taktéž zabývá předpovědí mechanických vlastností na základě teorií kaučukové elasticity. Bylo zjištěno, že výsledný materiál je částicový kompozit, kde se vzrůstajícím hmotnostním obsahem plniva v materiálu vzrůstá modul pružnosti. Predikce tahových křivek pomocí softwaru SEC.exe ukazovala pro reálný průběh křivky nereálné hodnoty hustot polymerních řetězců. U modelů, které vykazovaly reálné hodnoty hustot, byly naopak hodnoty napětí vyšší, než ve skutečnosti naměřené.
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Utilization of spent wood chips for biotechnological production of PHA
Ladický, Peter ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
The aim of this work was to study the suitability of wood shavings and sawdust as a substrate for microbial production of PHA by bacteria Burkholderia cepacia and Burkholderia sacchari. In the experimental part of the work the most appropriate approach of hydrolysis of wood shaving and sawdust and the effect of polyphenol and furfural concentration on ability to accumulate PHA was studied. Burkholderia sacchari had greater ability to accumulate PHA compared to Burkholderia cepacia. PHB values 87–89 % were achieved when Bulkholderia sacchari was cultivated on medium that contained detoxified hydrolysate of sawdust. Sawdust is therefore a promising substrate for microbial production of PHA in terms of reducing production costs and high content of PHB in biomass.
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Study of the medium influence on the pollutants sorption on microplastics
Hamplová, Marie ; Kučerík, Jiří (referee) ; Mravcová, Ludmila (advisor)
With the increasing production of plastic waste, the pollution of the environment by persistent microplastics is increasing simultaneously. Other environmental pollutants, such as various pharmaceuticals, can be sorbed on microplastics. This increases the toxicity of microplastics and their risk to living organisms. Nowadays, the use of so-called biodegradable polymers, which are supposed to degrade spontaneously, is expanding. However, their degradation can also produce microplastics. Therefore, this bachelor thesis deals with the sorption of drugs onto microplastics from biodegradable polyhydroxybutyrate (PHB) and with the influence of the surrounding aqueous environment on the rate and mechanism of sorption. Sorption of drugs onto microplastics made of synthetic polyethylene terephthalate (PET) was also carried out for comparison. The quantity of drugs was analysed by UPLC-ESI-TQ. Sorption was observed from the drug mixture for antibiotics tetracyclines. The pH value of the surrounding environment influenced the sorption rate of tetracyclines, the highest sorption occurred at acidic pH 4. According to the results of evaluation of sorption by Langmuir and Freundlich adsorption isotherms, PHB microplastics have a higher sorption capacity than PET microplastics. Therefore, PHB microplastics could be a risk for living organisms.
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Encapsulation of active substances into nanofibers and possibilities of their application
Procházková, Lucie ; Pernicová, Iva (referee) ; Skoumalová, Petra (advisor)
The master thesis was based on the optimization of the production of nanofiber covers and to gaine the product for subsequent functional use. The production of nanofiber covers was made by electrospinning and forcespinning from selected materials. Polyhydroxybutyrate, gelatin, chitosan and alginate were used as starting materials. After successful optimization, these materials were enriched with active ingredients ampicillin and ibuprofen for the functionalized use of covers for more effective wound healing. The theoretical part was focused on the issue of skin, healing processes, types of wounds and nanofibers, the characterization of selected starting materials for the formation of nanofibers was also mentioned. The practical part was based on the lengthy optimization of the preparation of fiber covers and later enriched with active ingredients. Furthermore, combined covers made of different materials with contents of both active ingredients were designed. This was followed by the characterization of all prepared covers from the point of view of stability in the short and long term. The gradual release of active ingredients was determined spectrophotometrically and by hifh performance liquid chromatography. It was also important to determine the antimicrobial activity of selected active substances. At the end of all testing, combined coatings containing both active ingredients were used for safety testing with human keratinocyte cells (HaCaT). Safety testing was based on determining the viability of human cells using the MTT test, to verify the LDH test. A scratch test was also performed, a wound healing test after the application of devised combined covers.
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Utilization of physico-chemical and spectroscopical methods in study on microbial extremofiles
Dobešová, Kateřina ; Enev, Vojtěch (referee) ; Sedláček, Petr (advisor)
The aim of this bachelor thesis is the determination od polyhydroxyalkanoates in the cells of extremophille bacterie, preformed by several methods. The chosen bacterium was Schlegelella thermodepolymerans DSM 15 344 Thermogravimetric analysis, infrared spectroscopy with Fourier transformation, and Raman spectroscopy were used in this thesis. The TGA method was used for the determination of extracellular and intracellular water in samples. The FTIR method was used in order to observe the amount od PHA, but aslo to observe the physical properties of the polymer in the sample, specifically to determine either amorphous or crystalline featuers of this polymer. Raman spectroscopy was used in oder to confirm the presence of PHA in the samples. All results e´were compared eith the results accomplished by gas chromatografy.
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Study of the interaction of selected pharmaceuticals with microplastics
Ioani, Aldina ; Řezáčová, Veronika (referee) ; Doležalová Weissmannová, Helena (advisor)
The bachelor thesis aims to study the interaction of selected pharmaceuticals from the group of analgesics and antibiotics with microplastics. The analysis was conducted in the mid-infrared region, which is used to identify chemical structure of the substances. Fourier transform infrared spectroscopy was used as an analytical method. The samples were subjected to technique attenuated total reflection. Ibuprofen and trimethoprim were chosen as analytes due to their common usage in the population. Sorption of pharmaceuticals was undergoing on polyhydroxybutyrate, which is a kind of biodegradable plastics and polyethene terephthalate, which is synthetic. Measured results showed that for the study of interaction with pharmaceuticals, it is more suitable to use biodegradable plastic polyhydroxybutyrate than polyethene terephthalate because of its better results. Infrared spectra output is very influenced by a selection of a solvent.
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Study on metabolism of polyhydroxyalkanoates in Rhodospirillum rubrum
Rubanová, Blanka ; Vodička, Juraj (referee) ; Slaninová, Eva (advisor)
This bachelor thesis deals with the study of polyhydroxyalkanoate (PHA) metabolism in Rhodospirillum rubrum under different cultivation conditions. The conditions differed in the aeration rate (aerobic, anaerobic, microaerobic) and in the presence of a light source (light cultivation, dark cultivation). First, different volumes of medium were suggested and tested for the fastest and the most suitable inoculum cultivation. Subsequently, cultures of Rhodospirillum rubrum under different conditions in terms of PHA content were compared by gas chromatography. In addition to gas chromatography, other complementary methods were used for better characterization of bacterial growth and metabolism, such as flow cytometry, UV-VIS spectroscopy (transmission and diffusion transmission modes). Fluorescence microscopy and cryo-scanning electron microscopy were used in terms of cell morphology. Furthermore, the ability of bacteria to synthesize color pigments (carotenoids and bacteriochlorophyll a) was investigated. The bacteria were cultivated on different types of media (liquid, solid), the color pigments were subsequently extracted, and their concentrations were calculated. UV-VIS spectroscopy, Raman spectroscopy and Fourier transform infrared spectroscopy (FTIR) were also used to measure color pigments.
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Properties of biocomposites based on flax fibers
Pelánová, Markéta ; Obruča, Stanislav (referee) ; Přikryl, Radek (advisor)
This thesis is focused on composites, which are composed of degradable components. The theoretical part deals with composite materials, treatment of fibers and properties of composites. The purpose of this study is to create a composite consisting of polyhydroxybutyrate, which is reinforced with flax fibers with various modifications. The thesis also evaluates the effect of reinforcing fibers in the composite material on the tensile properties and hydrolytic stability and degradability of the composite in compost in laboratory conditions.
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Controlled production of polyhydroxyalcanoates by bacterial strain Ralstonia eutropha using different substrates
Šnajdar, Ondřej ; Duroňová, Kateřina (referee) ; Márová, Ivana (advisor)
This bachelor’s thesis deals with production of polyhydroxyalkanoates (PHA) by bacterial strain Ralstonia eutropha. Production of PHA on different substrates has been studied to lower the costs of feedstock. In theoretical part the review has been done about the most important types of PHA, production strains and possibilities of cultivation of Ralstonia eutropha using different substrates. In practical part there has been studied production of PHA on different vegetable oils, including waste oils from different sources (restaurants, homes, food companies). Incorporation of different precursors for copolymer production control was studied too. The highest yields of poly-3-hydroxybutyrate (P3HB) have been achieved using waste oils. For economical and ecological reasons these oils are very suitable substrates. Using concentration 20 g/l of waste rapeseed oil from university canteen there has been produced 13,32 g/l of biomass containing 58,63% of P3HB in 84th hour of cultivation. The highest yield of PHA in precursors study has been achieved applying 1% propanol in 24th hour of cultivation. The PHA increase has been 97%. This PHA was composed from 91% of 3HB and 9% of 3-hydroxyvalerate.
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