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Production of PHB-HV from grape pomace by employing a pure culture of Cupriavidus necator
Jakešová, Michaela ; Kučera, Dan (referee) ; Kovalčík, Adriána (advisor)
This bachelor thesis deal with the production of PHB-HV copolymer employing a pure culture of Cupriavidus necator H16. First, Erlenmeyer flask´s cultivations using pure fructose as a source of carbon were performed. The mode of addition of valeric acid for the synthesis of PHB-HV was studied. Furthermore, the effect of stress factor (ethanol) on PHB-HV synthesis by this microorganism was investigated. The method obtained by optimization in Erlenmeyer flasks was applied to the synthesis of the PHB-HV copolymer in the bioreactor using fructose as a source of carbon, too. Furthermore, after enzymatic hydrolysis, a sugar extract from grape pomace was obtained as a source of carbon for the synthesis of PHB-HV. The fermentation in Erlenmeyer flasks and the bioreactor using an extract of grape pomace was carried out. Using this alternative carbon source 3.51 g/l PHB-HV copolymer was produced in Erlenmeyer flask, 5.06 g/l in bioreactor respectively. In the first case, the proportion of 3-hydroxyvalerate in the copolymer was 23.78 mol. % and in the second one, 9.39 mol. %.
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Production of selected metabolites by yeasts and algae cultivated under stress conditions
Mariničová, Veronika ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
The presented work was focused on the comparison between the production of selected metabolites by carotenogenic yeasts and microalgae cultivated under conditions of external stress. The main metabolites of interest were carotenoids, further lipophilic substances and lipids. Biotechnological overproduction of these metabolites could serve as a source of potentially beneficial substances not only for the pharmaceutical, cosmetic and food industries, but also for the production of third generation biofuels. Recently, there has been a growing interest in biofuels primarily from microalgae, which have a high potential in biofuel production and seem to be a promising source. The theoretical part deals with the description of individual genera of carotenogenic yeasts, microalgae, cyanobacteria, chemical composition of produced metabolites and brief biosynthesis. In addition, individual methods for analyzing the production of the metabolites of interest were described. The experimental part is focused on the comparison of production of carotenoids, coenzyme Q, ergosterols (phytosterols) and lipids by yeasts, microalgae and cyanobacteria. As a source of external stress, temperature, salt and light stress were chosen. The strains of Rhodotorula glutinis, Rhodotorula mucilaginosa, Sporidiobolus pararoseus and Cystofilobasidium macerans were studied from the yeast strains. Microalgae and cyanobacteria were Scenedesmus obliqus, Scenedesmus dimorphus, Chlorella sorokiniana, Chlorella saccharophila, Botryococcus brauni, Synechococcus nidulans and Arthrospira maxima. The yeast and algal strains were optimized for growth, carotenoid and lipid production. Applied salt stress showed a significant liquidation effect on algal and cyanobacterial strains. The thesis also monitored the biological stress, so-called co-cultivation of microalgae and yeasts. Further experiments will be the subject of future work.
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Interfacial rheology as the effective tool to description of interfacial behaviour of biofilms
Kachlířová, Helena ; Kráčalík, Milan (referee) ; Smilek, Jiří (advisor)
The aim of this diploma thesis is to optimize a method of interfacial rheology for testing the interfacial behaviour of biofilms on the liquid-air interface and after that use the method for studying the biofilm formation under optimal and stress conditions. For studying the biofilm formation, Kombucha was used. It is a microbial culture forming a cellulose biofilm on the interface. As the stress conditions, reduction of sucrose concentration, change of pH and change of ionic strength was used. Next, the ability of regeneration of biofilm formed on the interface was studied. The biofilm formation was occured in all cases except of increasing ionic strength. As expected, the best biofilm biofilm growth was observed under optimal condition, which means a sucrose concentration 100 g/l.
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Analysis of C. necator genome changes after evolutionary adaptation
Kroupa, Štěpán ; Obruča, Stanislav (referee) ; Brázda, Václav (advisor)
This bachelor’s thesis deals with analysis of mutations in bacterial populations of Cupriavidus necator H16 evolved in distinct stress conditions. This analysis was performed by processing data from the genome sequencing method „Next Generation Sequencing“, outsourced through the company DNALink. A list of mutations for each adapted population was constructed through bioinformatic methods. These mutations were then associated with specific areas of the reference Cupriavidus necator H16 genome from NCBI and analysed according to available information. Finally, the effect of these mutations on production of storage polymers polyhydroxyalkanoates was discussed.
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Long term sublethal stress in colonies of Saccharomyces cerevisiae deleted in WHI3
Krampotová, Ester ; Schierová, Michaela (advisor) ; Dušková, Michala (referee)
This diploma thesis is devoted to the topic of the development of natural non-pathogenic yeast colonies in non-laboratory diploid strain of Saccharomyces cerevisae with deletion WHI3, during long-term mild stress. The absence of Whi3 causes significantly higher senzitivity to medium composition relative to the parental strain, thus whi3Δ /whi3Δ colonies are a good model for study of the virulence induction due to environmental stress in pathogenic yeasts. Deletion of WHI3 in the BR-F strain results in a significant suppression of the ability to form structured colonies caused mainly by the reduced level of Flo11. The absence of Whi3 also has a negative effect on the expression of other genes involved in the stress response. The aim of the work is to determine whether changes in the expression of genes encoding stress proteins induced by sublethal doses of the inhibitor are dependent on Whi3, Yap6 and Mpt5 proteins. To induce stress in yeast, we used NaCl or CdSO4. The YAP6 and MPT5 genes encode regulatory proteins involved in the stress response, which expression is under control of Whi3. In whi3Δ /whi3Δ , the level of Hsp26, Tsa1, Pab1, and Gre2 was lower than in the parental strain. The Yap6 protein affects the response to the presence of Cd2+ in the medium, although according to SGD, only its...
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Autophagy as a mechanism of adaptation the yeast
Zieglerová, Leona ; Váchová, Libuše (advisor) ; Zikánová, Blanka (referee)
Autophagy is a degradation pathway, conserved from yeast to mammals. The uniqueness of this pathway lies in its function, it is applied in the cell especially under the adverse conditions. It helps the cell to deliver essential nutrients for life, it removes the damaged or superfluous organelles, protein aggregates and helps with recycling and maintains a constant inner environment. These functions can prolong cell life and the cells survive the adverse conditions. Autophagy may induce the programmed cell death type II. This paper describes the basic of autophagy machinery, regulation and influence of yeast autophagy to adapt to the stressful conditions. Understanding the mechanism and regulation of autophagy in yeast may help with the study of autophagy in mammals. In mammals, this degradation pathway disorders cause many diseases (especially neurodegenerative), autophagy also effects the formation of tumors. Powered by TCPDF (www.tcpdf.org)
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Degradation of the active pharmaceutical ingredient canagliflozin using electrochemical oxidation
Bolíková, Markéta ; Křížek, Tomáš (advisor) ; Kozlík, Petr (referee)
This bachelor thesis is aimed at degradation of the active pharmaceutical ingredient canagliflozin by the electrochemical oxidation. The aim is to explore the influence of given measurement conditions on the course and the results of the electrochemical oxidation and to find out, under which conditions the largest percentage of the degradation products is formed. These conditions are, for example, the concentrations and pH values of the buffers which are used to dissolve the samples of canagliflozin, the flow rate of the electrochemically stressed substance solution through the flow cell or the thickness of the gasket inserted in front of the working electrode. Canagliflozin is an orally administered blood glucose lowering antidiabetic and is used to treat diabetes mellitus 2. type. It is one of the inhibitors of the sodium-glucose transporter type 2. For the separation of the degradation products of this pharmaceutically active substance, a high performance liquid chromatography (HPLC) method with a UV/VIS diode array detector using an Agilent Poroshell 120 SB-Aq chromatographic column (2.1 × 100 mm; 2.7 µm) was chosen. The mobile phase contained an aqueous component, which was 10 mM formate buffer with pH 3.5 and the organic component, which was acetonitrile. The electrochemical oxidation of...
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Analysis of C. necator genome changes after evolutionary adaptation
Kroupa, Štěpán ; Obruča, Stanislav (referee) ; Brázda, Václav (advisor)
This bachelor’s thesis deals with analysis of mutations in bacterial populations of Cupriavidus necator H16 evolved in distinct stress conditions. This analysis was performed by processing data from the genome sequencing method „Next Generation Sequencing“, outsourced through the company DNALink. A list of mutations for each adapted population was constructed through bioinformatic methods. These mutations were then associated with specific areas of the reference Cupriavidus necator H16 genome from NCBI and analysed according to available information. Finally, the effect of these mutations on production of storage polymers polyhydroxyalkanoates was discussed.
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Production of PHB-HV from grape pomace by employing a pure culture of Cupriavidus necator
Jakešová, Michaela ; Kučera, Dan (referee) ; Kovalčík, Adriána (advisor)
This bachelor thesis deal with the production of PHB-HV copolymer employing a pure culture of Cupriavidus necator H16. First, Erlenmeyer flask´s cultivations using pure fructose as a source of carbon were performed. The mode of addition of valeric acid for the synthesis of PHB-HV was studied. Furthermore, the effect of stress factor (ethanol) on PHB-HV synthesis by this microorganism was investigated. The method obtained by optimization in Erlenmeyer flasks was applied to the synthesis of the PHB-HV copolymer in the bioreactor using fructose as a source of carbon, too. Furthermore, after enzymatic hydrolysis, a sugar extract from grape pomace was obtained as a source of carbon for the synthesis of PHB-HV. The fermentation in Erlenmeyer flasks and the bioreactor using an extract of grape pomace was carried out. Using this alternative carbon source 3.51 g/l PHB-HV copolymer was produced in Erlenmeyer flask, 5.06 g/l in bioreactor respectively. In the first case, the proportion of 3-hydroxyvalerate in the copolymer was 23.78 mol. % and in the second one, 9.39 mol. %.
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Interfacial rheology as the effective tool to description of interfacial behaviour of biofilms
Kachlířová, Helena ; Kráčalík, Milan (referee) ; Smilek, Jiří (advisor)
The aim of this diploma thesis is to optimize a method of interfacial rheology for testing the interfacial behaviour of biofilms on the liquid-air interface and after that use the method for studying the biofilm formation under optimal and stress conditions. For studying the biofilm formation, Kombucha was used. It is a microbial culture forming a cellulose biofilm on the interface. As the stress conditions, reduction of sucrose concentration, change of pH and change of ionic strength was used. Next, the ability of regeneration of biofilm formed on the interface was studied. The biofilm formation was occured in all cases except of increasing ionic strength. As expected, the best biofilm biofilm growth was observed under optimal condition, which means a sucrose concentration 100 g/l.
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