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Isolation, identification and characterization of extremophiles capable of PHA production
Vlasáková, Terézia ; Turková, Kristýna (referee) ; Obruča, Stanislav (advisor)
This diploma thesis is focused on isolation and identification of thermophilic microorganisms capable of production of polyhydroxyalkanoates (PHA) in the sample of activated sludge from wastewater treatment. 6 culture samples were isolated from activated sludge by means of cultivation technics and methods of molecular biology. They were closer specified by comparing nucleotide sequences of 16S-rRNA gene and assigned to bacterial genus Anoxybacillus. The production of PHA by this genus was not reported in literature so far. Samples were confirmed to contain phaC gene that codes the enzyme PHA-synthase and they also gave a positive response to staining colonies with Nile red, what refers to presence of intracellular lipidic structures. However, the PHA production by isolates was not successful. The reason should be an inappropriate production medium or conditions. The positive phenotype result of Nile red dyeing was probably achieved by production of huge amount of lipids by bacterial cells that provides similar fluorescence than PHA granules.
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Production of polyhydroxy alkanoates from selected waste substrates
Vlasáková, Terézia ; Obručová,, Hana (referee) ; Obruča, Stanislav (advisor)
This bachelor's thesis is focused on study of polyhydroxyalkanoates (PHA) production by bacterial strains Burkholderia sacchari and Burkholderia cepacia. It also deals with influence of various factors on their production. The main studied area was PHA production from substrate prepared from corn stover. In theoretical part a brief description of PHA was given. We also summarized options on their practical applications and possibilities of their bacterial production from waste substrates. In experimental part of work the impact of used precursors on biomass growth, PHA production and incorporation of 3HV monomer into the polymer chain was monitored. Further, a method of corn material hydrolysis was optimized. Bacterial cultures were cultivated on prepared hydrolysates. The best results in cultivation on hydrolysates of corn silage were achieved when the hydrolysis was performed by combination of chemical and enzymatic way. Cultivations of both bacteria were also transferred into the laboratory bioreactor.
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Isolation, identification and characterization of extremophiles capable of PHA production
Vlasáková, Terézia ; Turková, Kristýna (referee) ; Obruča, Stanislav (advisor)
This diploma thesis is focused on isolation and identification of thermophilic microorganisms capable of production of polyhydroxyalkanoates (PHA) in the sample of activated sludge from wastewater treatment. 6 culture samples were isolated from activated sludge by means of cultivation technics and methods of molecular biology. They were closer specified by comparing nucleotide sequences of 16S-rRNA gene and assigned to bacterial genus Anoxybacillus. The production of PHA by this genus was not reported in literature so far. Samples were confirmed to contain phaC gene that codes the enzyme PHA-synthase and they also gave a positive response to staining colonies with Nile red, what refers to presence of intracellular lipidic structures. However, the PHA production by isolates was not successful. The reason should be an inappropriate production medium or conditions. The positive phenotype result of Nile red dyeing was probably achieved by production of huge amount of lipids by bacterial cells that provides similar fluorescence than PHA granules.
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Production of polyhydroxy alkanoates from selected waste substrates
Vlasáková, Terézia ; Obručová,, Hana (referee) ; Obruča, Stanislav (advisor)
This bachelor's thesis is focused on study of polyhydroxyalkanoates (PHA) production by bacterial strains Burkholderia sacchari and Burkholderia cepacia. It also deals with influence of various factors on their production. The main studied area was PHA production from substrate prepared from corn stover. In theoretical part a brief description of PHA was given. We also summarized options on their practical applications and possibilities of their bacterial production from waste substrates. In experimental part of work the impact of used precursors on biomass growth, PHA production and incorporation of 3HV monomer into the polymer chain was monitored. Further, a method of corn material hydrolysis was optimized. Bacterial cultures were cultivated on prepared hydrolysates. The best results in cultivation on hydrolysates of corn silage were achieved when the hydrolysis was performed by combination of chemical and enzymatic way. Cultivations of both bacteria were also transferred into the laboratory bioreactor.
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