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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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Evolutionary engineering of cyanobacteria with respect to PHA accumulation
Vašířová, Kristýna ; Slaninová, Eva (referee) ; Samek, Ota (advisor)
The aim of this diploma thesis was to subject selected cyanobacterial strains to adaptive evolution and subsequently investigate the properties of the resulting adapted strains, especially their changes related to polyhydroxyalkanoates (PHA) accumulation. The theoretical part of the work describes in more detail the issue of cyanobacteria, PHA and their interconnection in the field of evolutionary engineering. Cyanobacterial strains Synechocystis sp 6803 and. Synechocystis salina CCALA 192 were used for evolutionary experiments. Selection pressures of hydrogen peroxide and copper. were applied to selected representatives. The resulting cultures and their ability to accumulate PHA were subsequently assessed by gas chromatography. Both of these selection pressures proved to be unsuitable, as strong growth inhibition was observed after their application to cultures, which did not allow the application of evolutionary engineering methods. In the second half of the experimental part, the provided adapted strains to 6% NaCl were monitored. Adaptation has been shown to have a positive effect on microorganisms, as they have a higher biomass content, better stress resistance and a slight increase in PHA accumulation.
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Evolutionary engineering of cyanobacteria with respect to PHA accumulation
Vašířová, Kristýna ; Slaninová, Eva (referee) ; Samek, Ota (advisor)
The aim of this diploma thesis was to subject selected cyanobacterial strains to adaptive evolution and subsequently investigate the properties of the resulting adapted strains, especially their changes related to polyhydroxyalkanoates (PHA) accumulation. The theoretical part of the work describes in more detail the issue of cyanobacteria, PHA and their interconnection in the field of evolutionary engineering. Cyanobacterial strains Synechocystis sp 6803 and. Synechocystis salina CCALA 192 were used for evolutionary experiments. Selection pressures of hydrogen peroxide and copper. were applied to selected representatives. The resulting cultures and their ability to accumulate PHA were subsequently assessed by gas chromatography. Both of these selection pressures proved to be unsuitable, as strong growth inhibition was observed after their application to cultures, which did not allow the application of evolutionary engineering methods. In the second half of the experimental part, the provided adapted strains to 6% NaCl were monitored. Adaptation has been shown to have a positive effect on microorganisms, as they have a higher biomass content, better stress resistance and a slight increase in PHA accumulation.
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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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