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Study on biotechnological potential of Pseudomonas thermotolerans
Vašířová, Kristýna ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
The aim of this bachelor‘s thesis was to evaluate biotechnological potential of the bacterium Pseudomonas thermotolerans. The ability of bacterium to produce polyhydroxyalkanoates (PHA) and biosurfaktants was examined. Theoretical part deals with characteristic of extremophilic bacteria, PHA and biosurfactants. In the experimental part, the bacterium was tested for the presence of gene that catalyzes the synthesis of mcl-PHA (phaC1), using molecular technique PCR. Presence of the gene was confirmed by the experiment. Based on the result, PHA production conditions were optimized, such as selection of ideal media for production, suitable cultivation temperature and precursors. However, non of the tests confirmed the ability of production. Another of the experiments was to examine the bacterium’s ability to produce biosurfactants. Testing was realized in a suitable mineral medium with different carbon sources. Demonstrable production of biosurfactants was only on mineral medium supplemented by yeast extract and fructose. The thin layer chromatography (TLC) demonstrated, that the bakterium produces a mixture of monorhamnolipids and dirhamnolipids. However, higher portion is represented by dirhamnolipids.
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Characterization of PHA producing microbial cells by advanced microscopic and cytometric techniques
Dlouhá, Karolína ; Nováčková, Ivana (referee) ; Obruča, Stanislav (advisor)
The aim of this bachelor thesis is to document production of polyhydroxyalkanoates in selected bacterial strains, which were Pseudomonas thermotolerans, Chelatococcus daeguensis, Tepidiphilus thermophilus and Chelatococcus thermostellatus. In the case of the microorganisms Tepidiphilus thermophilus and Pseudomonas thermotolerans, the production of PHA´s has not yet been described, and in the Chelatococcus bacteria, which were analysed in this work, the production has not yet been documented by electron microscopy. In this work, the selected producers were analysed by flow cytometry first, using BODIPY and Nile Red as fluorescence probes. Selected producers, for which was production confirmed, were analysed by other methods, such as fluorescence microscopy, cryo scanning electron microscopy and transmission electron microscopy. In the bacterial culture of Pseudomonas thermotolerans, PHA´s production wasn´t confirmed by the first analysis by flow cytometer. For other microorganisms was production confirmed. Chelatococcus bacteria clearly proved to be better producers. Bacterial cells of Tepidiphilus thermophilus produced smaller granules and in lower amount.
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Characterization of PHA producing microbial cells by advanced microscopic and cytometric techniques
Dlouhá, Karolína ; Nováčková, Ivana (referee) ; Obruča, Stanislav (advisor)
The aim of this bachelor thesis is to document production of polyhydroxyalkanoates in selected bacterial strains, which were Pseudomonas thermotolerans, Chelatococcus daeguensis, Tepidiphilus thermophilus and Chelatococcus thermostellatus. In the case of the microorganisms Tepidiphilus thermophilus and Pseudomonas thermotolerans, the production of PHA´s has not yet been described, and in the Chelatococcus bacteria, which were analysed in this work, the production has not yet been documented by electron microscopy. In this work, the selected producers were analysed by flow cytometry first, using BODIPY and Nile Red as fluorescence probes. Selected producers, for which was production confirmed, were analysed by other methods, such as fluorescence microscopy, cryo scanning electron microscopy and transmission electron microscopy. In the bacterial culture of Pseudomonas thermotolerans, PHA´s production wasn´t confirmed by the first analysis by flow cytometer. For other microorganisms was production confirmed. Chelatococcus bacteria clearly proved to be better producers. Bacterial cells of Tepidiphilus thermophilus produced smaller granules and in lower amount.
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Study on biotechnological potential of Pseudomonas thermotolerans
Vašířová, Kristýna ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
The aim of this bachelor‘s thesis was to evaluate biotechnological potential of the bacterium Pseudomonas thermotolerans. The ability of bacterium to produce polyhydroxyalkanoates (PHA) and biosurfaktants was examined. Theoretical part deals with characteristic of extremophilic bacteria, PHA and biosurfactants. In the experimental part, the bacterium was tested for the presence of gene that catalyzes the synthesis of mcl-PHA (phaC1), using molecular technique PCR. Presence of the gene was confirmed by the experiment. Based on the result, PHA production conditions were optimized, such as selection of ideal media for production, suitable cultivation temperature and precursors. However, non of the tests confirmed the ability of production. Another of the experiments was to examine the bacterium’s ability to produce biosurfactants. Testing was realized in a suitable mineral medium with different carbon sources. Demonstrable production of biosurfactants was only on mineral medium supplemented by yeast extract and fructose. The thin layer chromatography (TLC) demonstrated, that the bakterium produces a mixture of monorhamnolipids and dirhamnolipids. However, higher portion is represented by dirhamnolipids.
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