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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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Removal of selected xenobiotics at municipal WWTP´s
Žižlavská, Adéla ; Rusník,, Igor (referee) ; Bodík,, Igor (referee) ; Hlavínek, Petr (advisor)
The development of the chemical and pharmaceutical industry and the massive use of synthetic substances in various industries has become indispensable for our society. Unfortunately, this progress has brought a whole new kind of pollution. It is a group of substances which imagine for living organisms’ greater risk than 'normal' organic pollution. Because they have extraneous origin compared with naturally occurring compounds, they are called "Xenobiotics." It is a group comprising a large number of different substances, with dissimilar physical, chemical and biological properties such as pharmaceuticals, pesticides, detergents, paints, varnishes, plastic packaging, food additives, cosmetic products, drugs, and many others. Although these substances began to enter the water cycle as early as 30 years of the 20th century, their concentration often moves to the limit of the detection limit, so their presence in the water began to be examined in more detail in the 1980s, following the development of analytical technologies chemical composition of water. The main danger of these substances is that doesn’t primarily effect on water quality, but directly affect on the organisms by confusing of the endocrine system or by directly damaging RNA or DNA, causing mutagenic changes, cancer-causing proliferation, birth defects, metabolic disorders, gender changes, the degeneration of reproductive capacity of entire populations or the disappearance of self-preservation instinct. The source of xenobiotics in the water cycle is predominantly wastewater, which generated through the wastewater treatment, which not designed to eliminate this type of pollution. he dissertation thesis deals with the issue of xenobiotics in wastewater and is divided into the theoretical and experimental part. The theoretical part summarizes the xenobiotics current state of knowledge in wastewater from their history, origin, classification, possibilities of elimination to impacts on the environ
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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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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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Responses of Rhodococcus strains to heavy metal stress
Volf, Jan ; Štěpánek, Václav (advisor) ; Martínková, Ludmila (referee)
Increasing heavy metal pollution can be dealt with not only through physical and chemical decontamination processes but also utilizing microbial bioremediation, a much more environmentally friendly way. Members of genus Rhodococcus are suitable candidates for effective bioremediations of heavy metals due to their considerable adaptability and extreme resistance to different stress conditions, including those related to presence of heavy metals or their toxic compounds. Individual rhodococcal strains are more resistant to heavy metals than most of other microorganisms and they are capable of biotransformating them to less toxic forms or at least of accumulating effectively or adsorbing them on produced polysaccharides and specific surface active substances. Their bioremediation potential is very high even though, so far, only molecular mechanisms of rhodococcal resistances to toxic arsenic compounds have been studied more in detail.
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Removal of selected xenobiotics at municipal WWTP´s
Žižlavská, Adéla ; Rusník,, Igor (referee) ; Bodík,, Igor (referee) ; Hlavínek, Petr (advisor)
The development of the chemical and pharmaceutical industry and the massive use of synthetic substances in various industries has become indispensable for our society. Unfortunately, this progress has brought a whole new kind of pollution. It is a group of substances which imagine for living organisms’ greater risk than 'normal' organic pollution. Because they have extraneous origin compared with naturally occurring compounds, they are called "Xenobiotics." It is a group comprising a large number of different substances, with dissimilar physical, chemical and biological properties such as pharmaceuticals, pesticides, detergents, paints, varnishes, plastic packaging, food additives, cosmetic products, drugs, and many others. Although these substances began to enter the water cycle as early as 30 years of the 20th century, their concentration often moves to the limit of the detection limit, so their presence in the water began to be examined in more detail in the 1980s, following the development of analytical technologies chemical composition of water. The main danger of these substances is that doesn’t primarily effect on water quality, but directly affect on the organisms by confusing of the endocrine system or by directly damaging RNA or DNA, causing mutagenic changes, cancer-causing proliferation, birth defects, metabolic disorders, gender changes, the degeneration of reproductive capacity of entire populations or the disappearance of self-preservation instinct. The source of xenobiotics in the water cycle is predominantly wastewater, which generated through the wastewater treatment, which not designed to eliminate this type of pollution. he dissertation thesis deals with the issue of xenobiotics in wastewater and is divided into the theoretical and experimental part. The theoretical part summarizes the xenobiotics current state of knowledge in wastewater from their history, origin, classification, possibilities of elimination to impacts on the environ
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Composting of materials contaminated by micropollutants
Kočí, Petra ; Cajthaml, Tomáš (advisor) ; Innemanová, Petra (referee)
Micropollutants are compounds of anthropogenic origin, detected at concentrations of ng·g-1 - mg·g-1 in the environment. Micropollutants are synthetic or natural compounds. Micropollutants are released into the environment due to various human activities. Despite low concentrations in the environment, they represent a real threat to organisms, because of their specific biological effects. Sewage sludge is used in agriculture due to its hight nutritious content. Via using sludge in agriculture, contained micropollutants can be released to environment. Composting is a cheap bioremediation method, used from 80's of the last century. This diploma thesis is focused on composting of sewage sludge, contamined with micropollutants. High degradation rate was observed in artificially contaminated sludge, mainly with endocrine disruptors (up to 100 %). Lower degradation rate was observed in sewage sludge from a pharmaceutical plant, some of the compounds were not degraded at all.
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