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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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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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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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|
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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Metamorphoses of \kur{Borrelia burgdorferi} sensu lato spirochetes: from dormant to motile forms
MORÁVKOVÁ, Veronika
The aim of the study was to obtain and elaborate information focused on metamorphoses of Borrelia burgdorferi sensu lato (s.l.) spirochetes. The research included detection of various stress conditions for production of dormant forms of Borrelia burgdorferi s.l. spirochetes in vitro. Subsequently, metamorphoses from dormant to motile stages was observed under favourable conditions. Proper PCR method for primers aimed to detect dormant forms of Borrelia was delevoped. The infectious potential of dormant spirochetes of Borrelia burgdorferi sensu stricto (s.s.) complex was observed in vivo. Transformations of spirochetes have also been observed in real time and individual stages have been recorded.
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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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