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Utilization of PHA producing bacteria in bioremediation technologies
Šuráňová, Zuzana ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
The aim of this work is study of utilization of PHA producing bacteria in bioremediation technologies. For this study were used bacteria Pseudomonas putida KT2440 and two isolates from soil contaminated by petroleum - Pseudomonas gessardii (D2) a Pseudomonas fulva (D3). The experimental part describes especially study of feather biodegradation using selected microbial strains. All the tested bacterial strains were capable of feather degradation and utilization as the sole carbon source. During biodegradation experiment, we monitored weight loss of feather, protease and keratinase activity, concentration of bacterial biomass and PHA content as well as pH. The highest biodegradation ability and keratinase activity was observed in Pseudomonas putida. None of tested bacteria accumulated detectable amount of PHA during growth on waste feather, nevertheless, bacterial biomass grown during feather degradation can be used as an inoculum for PHA production on waste frying oil and octanoic acid. Using this experimental setup, high PHA content (54% of cell dry weight) was achiaved in Pseudomonas putida. Another part of the thesis deals with biodegradation of petroleum oil. The highest capability of growth on this carbon source were determined in Pseudomonas fulva.
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Enzymes of Candida tropicalis yeast biodegrading phenol
Koubková, Zuzana ; Stiborová, Marie (advisor) ; Turek, Michal (referee)
Effluents of industrial wastewaters from oil refineries, paper mills, dyes, ceramic factories, resins, textiles and plastic contain high concentrations of aromatic compounds, which are toxic to organisms. Degradation of these compounds to tolerant limits before releasing them into the environment is an urgent requirement. Candida tropicalis yeast is an important representative of eucaryotic microorganisms that are able to utilize phenol. During the first phase of phenol biodegradation, cytoplasmatic NADPH-dependent phenol hydroxylase of C. tropicatis oxidizes phenol to catechol. Catechol is in the second phase of biodegradative process oxidized to cis,cis-muconic acid by the reaction catalyzed with catechol-1,2-dioxygenase. In this diploma thesis we investigated the effect of the heavy metal ions on NADPH-dependent phenol hydroxylase and catechol-1,2-dioxygenase of C. tropicalis. Phenol hydroxylase was inhibited by Cu2+ and Pb2+ ions. Catechol dioxygenase was inhibited by all substances containing heavy metal ions (Fe2+ , Mn2+ , Cd2+ , Cu2+ and Pb2+ ), which were tested in this work. The most effective inhibition was produced by Pb2+ followed by Mn2+ , Cd2+ Fe2+ and Cu2+ ions. The higher sensitivity of catechol-1,2-dioxygenase to heavy metal ions might follow from the presence of histidine residue...
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Biodegradation of polycyclic aromatic hydrocarbon in wood treated with creosote oil
Fabiánová, Tereza ; Cajthaml, Tomáš (advisor) ; Gabriel, Jiří (referee)
Wood contaminated by polycyclic aromatic hydrocarbons (PAH) represent a serious environmental risk. Composting seems to be an effective method for decontamination of such waste material. PAHs are degraded mainly under aerobic conditions the efficiency of composting could be strongly affected by the process conditions. This work is aimed at the comparison of PAH removal by a 340-days-long composting in laboratory scale composting pilots and a 240-days-long process in a pilot scale. To set up composting piles a "green substrate" commercially used for button-mushroom production and a grass substrate were used. The use of the grass substrate led to a higher PAHs removal (97% of the initial sum of PAHs) in comparison to the green substrate (81% of the initial sum of PAHs). Further, the effect of the size of wood particles on the PAHs removal efficiency was demonstrated. Analysis of phospholipid fatty acids showed an enhanced growth of gramm-negative bacteria and fungi during the composting processes. During the degradation process the enzyme activity of laccase was detected. The ecotoxicological test using a bioluminescent bacteria Vibrio fischeri and a seed germination test using seeds of Hordeum vulgare L. showed that probably no toxic products were formed during the PAHs degradation in these experiments.
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Phylogenetic analysis of Rieske dioxygenases large subunits genes in soil contaminated with jet fuel
Ptáček, Jakub ; Bořek Dohalská, Lucie (advisor) ; Poljaková, Jitka (referee)
The former military air-base Hradcany is among the most contaminated with organic pollutants localities in Czech Republic. Main cleanup strategy in the area is the bioremediation taking advantage on the natural potential of the autochthonous soil microorganisms to evolve catabolic pathways for in situ degradation of the pollutant. The diversity and abundance of the pathways, as well as the specificity and activity of the encoded enzymes are priority biotic factors determining the bioremediation efficiency. Main task of this work was to analyze the bacterial diversity in jet fuel contaminated soils based on key catabolic genes encoding the Rieske non-haem iron dioxygenases of the toluene/ biphenyl oxygenase branch. High molecular soil DNA was extracted and the sequences encoding catabolic genes were selectively enriched by hybridization to biotinylated oligonucleotides on magnetic microbeads with covalently bound streptavidin. Fragments of the genes for the -subunits of Rieske non-haem iron oxygenases were amplified and analyzed by restriction analysis, cloning and sequencing. Their evolutionary histories were inferred using the Neighbour-Joining and the maximum likelihood methods. The catabolic genes diversity in the actively bioremediated and highly polluted soil HRB was compared with the diversity in the...
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Study of bioremediation of some organic polutants by Cupravidus necator
Miléřová, Miluše ; Hurtová, Jana (referee) ; Obruča, Stanislav (advisor)
The aim of this work is the study of bioremediation of p-nitrophenol using bacteria Cupriavidus necator. The theoretical part contains a summary of organic pollutants and the bioremediation aproaches used to remove them from the environment. In the experimental part of this work we used bacteria Cupriavidus necator H16 and its mutant strain Cupriavidus necator H16/PHB-4 unable of PHB accumulation. The experiments were performed with different concetrations of p-nitrophenol and also under none-optimal conditions. Apart from concentration of p-nitrophenol, viability of the bacterial cultures was determined by flow cytometry. During cultivation in progress for sub-optimal conditions with a concentration of 50 mg•l-1 p-nitrophenol at 4°C using a flow cytometer seemed more resistant bacteria Cupriavidus necator H16. On the contrary strain Cupriavidus necator H16/PHB-4 was more resistant at 37 °C. The biodegradation assay was also performed in fermentor, during 51 hours lasting cultivation, bacterial culture degraded 65 % of p-nitrophenol and moreover decreased wavelenght of maximum in absorption spectra from 401 m to 384 nm was observed. The quantity of PHB was analyzed by gas chromatography with FID detector along with the p-nitrophenol determined by spectrophotometry, whose maximum content of PHB was achieved in 20 hour of cultivation
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Influence of soil properties on mobility of selected risk elements in the soil and some methods of their decontamination
BEDRNÍČEK, Jan
Development of society and the increasing population of the planet are related to the increasing discharge of wastes and pollutants. These include heavy metals. Purpose of this bachelor´s thesis is to briefly clarify behaving of lead, cadmium and mercury in the soil; it means content, forms, mobility, sorption and its influencing factors and their impact on the animal and plant organism. This thesis also gives a wider view on the issue of decontamination techniques with focusing on bioremediation and mainly phytoremediation with the use of plant hyperaccumulators.
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