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Modification of porosity of bacterial cellulose in situ
Ondruchová, Barbora ; Pernicová, Iva (referee) ; Kovalčík, Adriána (advisor)
This bachelor thesis deals with the production and modification of porosity of bacterial cellulose in situ using the bacterium Komagataeibacter xylinus. The theoretical part of the work was focused on the review of various methods of culturing Komagataeibacter xylinus and the production of porous samples of bacterial cellulose. The sizes of pores in bacterial cellulose depend mainly on the applied cultivation method. Bacterial cellulose produced statically or dynamically contains pores with the dimensions of approximately 0.02 µm to 10 µm. The difference in porosity in bacterial cellulose prepared by static and dynamic cultivation was confirmed experimentally. The production yields of bacterial cellulose were compared and discussed. Next, the porosity of the bacterial cellulose was modified in situ by the addition of wax particles. Scanning electron microscopy confirmed, that the accumulation of wax particles in the production medium could significantly support the porosity of bacterial cellulose and, at the same time, increase its production.
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Analysis of adherent cells confluency in 2D culture
Bracková, Michaela ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
First part of this semester work dealing with the theoretical description of adherens cells, particularly structure and functions of this cells. Subsequently work is devoted to description of cell culturing, with mention of conditions which are necessary for cell culturing, following by substances which promote cell growth. Last part of theoretical research is concern with microscopy technique that is suitable for studying of adherent cells. Subsequently it is about fluorescence and confocal microscopes. Practical part dealing with cell culturing of adherent cells and the evaluation of realized experiments.
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Usage of flue gas as a source of CO2 for algae cultivation in bioreactors
Slonek, Jaroslav ; Juřena, Tomáš (referee) ; Brummer, Vladimír (advisor)
This master`s thesis is focused on possibility of use of flue gas as source of CO2 for algae cultivation in bioreactors. The first part of the master`s thesis is devoted to a qualitative and quantitative description of flue gas sources as CO2 source for algae cultivation. In the thesis industry sectors with the largest share of CO2 emissions on a global scale and in the Czech republic are summarized. On the basis of these data it was possible to determine cheap sources of CO2 for algae cultivation producing valuable biomass and other bio-products. The composition of flue gas from these sources was discussed in this section also with given expected outlook within the following years. The next part of the thesis is devoted to the influence of flue gas properties on algae cultivation. The impact of flue gas individual parameters on cell growth rate, quality and composition of biomass is described there. The description and design of the bioreactor is processed in the next part of the thesis. The experimental part of the thesis was performed on a bioreactor with similar properties. The next part of the thesis describes the effect of light on algae cultivation and the design of a regulation scheme for bioreactor light source. The experimental part of the thesis describes methods used to determine the concentrations of flue gas substances that could pass from flue gas into cultivation medium for algae cultivation to measure the changing composition of the flue gas in the closed experimental bioreactor system. The processed data from experimental measurements are presented and are discussed. The change in the composition of the culture medium for algae (without the presence of algae) due to flue gas aeration was monitored in the experimental part. It includes also the measurement of the change in the composition of the flue gas in time. All obtained and collected information are then summarized and discussed in the final part of the master`s thesis.
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Study of oligandrin protein secreted by oomycete Pythium oligandrum
Neykulova, Anastasia ; Bělonožníková, Kateřina (advisor) ; Adámková, Lyubina (referee)
Oomycete Pythium oligandrum acts as a mycoparasite of pathogenic fungi, bacteria and oomycetes in soil due to the production of a number of hydrolytic enzymes for the degradation of polysaccharides and proteins in the cell wall of the host. At the same time, P. oligandrum can interact with plant roots through specific elicitors and stimulate plant defense and growth. Thus, P. oligandrum is a successfully used environmentally friendly biological control agent of plants. Among the secreted elicitors P. oligandrum to the low molecular weight proteins belong oligandrins (~10 kDa), which have a characteristic structure and a conserved sequence among the group of so-called elicitins, and at the same time are not homologous to plant proteins. In the framework of this bachelor thesis, was analyzed the production of oligandrin in various types of growing media after cultivation of P. oligandrum. The total proteolytic activity and the content of phenolic substances as other possible elicitors of plant defense reactions were also observed in culture media. Further, this work focused on the possibility of recombinant oligandrin expression in E. coli and its subsequent purification. Key words: Pythium oligandrum, oligandrin, elicitors, cultivation, recombinant expression [IN CZECH]
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Vliv jarní kultivace ozimé pšenice na její výnos a zaplevelení
Novák, Radim
This bachelor thesis is focused on the impact of spring cultivation on the cover of the winter wheat, its number of straws and the weed infestation. A small field experiment was performed for an assessment. There were 10 smaller parcels of land picked out for this experiment. The first 5 parcels of land were used for the testing of different types of cultivation: rotary hoe + tine harrow, rotary hoe + meadow harrow, rotary hoe, tine harrow, meadow harrow. The other 5 parcels of land were used for the control. These factors were monitored: the number of straws, yield and the number of weed individuals. The number of straws was affected the most by the combination of the rotary hoe and the tine harrow. The number of the weed individuals per 1 m2 was reduced the most by the combination of the rotary hoe and the tine harrow. The lowest efficacy of the weed regulation was measured by the meadow harrow. Thanks to this data, the machines for the spring cultivation can be well organized and used.
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Human Dental Pulp Stem Cells Cultured in Xenogeneic-Free Supplemented Media
Suchánková Kleplová, Tereza ; Jouklová, Nela (advisor) ; Merglová, Vlasta (referee) ; Bartoňová, Marie (referee)
Human dental pulp stem cells cultured in xenogeneic-free supplemented media Summary Introduction: The topic of the study is the cultivation of dental pulp stem cells (hDPSCs) in a xenogeneic-free culture medium. It is not permissible to use cells upon growing under the influence of xenogeneic (extraneous) substances in human clinical practice. The most frequently used in cultivation of hDPSCs is fetal calf serum (FCS/FBS). Unfortunately, these supplements are widespread in hMSCs cultivation, and all gold standard hMSCs properties were postulated in cells cultivated using these supplements. This raises the basic question if and how xenogeneic blood derivatives affect the properties of cells and their growth characteristics. There are two options for replacing these xenogeneic substances in the culture medium: the so-called serum-free media, or human blood supplements, ideally autologous ones. The conducted research was aimed at identifying the effects of xenogeneic and human blood supplements on basic hDPSCs characteristics that are fundamental to introduce the cell therapy into regular medical practice. Method: By culturing 12 hDPSC lines obtained from adult, deciduous, and natal teeth in 12 different culture media, we investigated the effect of FCS, human blood derivatives, i.e., blood plasma (HP), and...
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Microalgae harvesting
Drozd, Jiří ; Naď, Martin (referee) ; Lošák, Pavel (advisor)
This work describe microalgae and their properties. Within the scope of the work, a research was conducted on the utilized microalgae and the possibilities of cultivation in the Central European region were examined. Furthermore, harvesting and drying methods for microalgae were analyzed, and an experiment was conducted using a promising spray drying method. The research revealed the properties and potential uses of microalgae. Various types of microalgae, their nutritional values, and possibilities of application in a wide range of fields were investigated. The cultivation possibilities of microalgae in the Central European region, including the utilization of wastewater and renewable energy sources, were also examined. Harvesting and drying methods for microalgae were analyzed, considering the advantages and disadvantages of different techniques such as flocculation, centrifugation, and spray drying. An experiment utilizing the spray drying method was conducted, and the results showed that this method is effective and enables the achievement of high quality and purity of dried microalgae. The work also explores the possibilities of further development and utilization of microalgae. It describes the potential use of microalgae as food supplements and as alternatives to traditional protein sources. Additionally, the potential applications of microalgae in the pharmaceutical and cosmetic industries were mentioned. In conclusion, this work provides information on the utilization of microalgae and the methods of cultivation, harvesting, and drying. The results suggest that microalgae have great potential for further development and utilization in various industries.
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The influence of Biochar Cultivation in Soil on its Physicochemical Characteristics
Hladký, David ; Sovová, Šárka (referee) ; Kalina, Michal (advisor)
This bachelor thesis is focused on the effect of soil factors on the physical-chemical characteristics of biochar. It describes the change in biochar after a long-term cultivation experiment, as well as, sequential leaching that was implemented on a shorter time scale. The theoretical part describes individual physical-chemical characteristics, structure, possible applications and optimal methods of biochar characterization. In the thesis, we have learnt why biochar is called a soil conditioner, not a fertilizer. The experimental part deals with the study of changes in the physicochemical characteristics of biochar after the cultivation experiment. These results are compared with the sequential leaching of biochar in selected media (water, citric acid, hydrogen peroxide) that simulate the effects that can act on biochar in soil. The experimental results show the similarity of individual observed values of the samples after the long-term cultivation experiment and sequential extractions. It can be seen that the alkaline nature of biochar is caused by salts of alkali metal and alkaline earth metal. Mostly due to the presence of potassium in the biochar structure and then also the presence of calcium and sodium. These ions are also the explanation of the high conductivity, which together with pH, decreases significantly after five leaching cycles, and stabilize after a two-year cultivation period. All biochars samples used in the work have a high content of organic matter, which is a optimal prerequisite for the use in agriculture and the improvement of soil properties.
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Optimization of culture conditions of selected {E.coli} strains producing recombinant proteins
MICHALCOVÁ, Zuzana
Escherichia coli is one of the most preferred organisms to produce recombinant proteins. E. coli is a well-established host that offers ease of genetic manipulation, short culture times, continuous fermentation capability and affordable culture media. To achieve high levels of protein expression, and therefore high recombinant protein production, optimization of culture conditions is required. This thesis is focused on the optimization of cultivation, the practical part is focused on the production of Uridine Phosphorylase (UP) and Purine Nucleoside Phosphorylase (PNP) enzymes. E. coli bacteria producing UP or PNP were cultured on six media with different nutrient sources. Protein production was verified on a spectrophotometer by the Bradford assay and further by polyacrylamide gel electrophoresis with SDS (SDS-PAGE). Enzyme activity was measured by HPLC assay. In addition, the dry weight biomass was determined for all samples. Based on results from these assays, Terrific Broth medium with glucose addition after 4 h of cultivation (TBG4) was selected and subsequently used for upscaling to the fermenter. Successful cultivation in the fermenter confirmed the results of cultivation in Erlenmeyer flasks.
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