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Biotechnological production of PHA by selected bacterial isolates
Matějka, Filip ; Šedrlová, Zuzana (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis deals with the production of polyhydroxyalkanoates (PHA) using thermophilic bacterial isolates with designation 34, 35 and BŽ. Bacterial straines were isolated from activated sludge and compost The theoretical part contains a description of PHA, extremophilic bacteria and detection techniques for the determination of polyhydroxyalkanoates. In the experimental part, the presence of the phaC gene, which is crucial for the ability to produce PHA, was first determined by PCR and bacterial strains were also subjected to DNA sequencing of 16S rRNA gene which enabled preliminary taxonomical classification of the isolates. In the next part of the experimental work, the conditions for biomass growth and PHA production were optimized. Suitable carbon source, the ideal temperature for cultivation and the influence of precursors on the production of copolymers were studied and identified. The composition and proportion of PHA were determined spectrophotometrically and by GC-FID. Finally, visual screening of PHA accumulation inside bacterial cells was performed using fluorescence microscopy.
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Biotechnological production of PHA by selected bacterial isolates
Matějka, Filip ; Šedrlová, Zuzana (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis deals with the production of polyhydroxyalkanoates (PHA) using thermophilic bacterial isolates with designation 34, 35 and BŽ. Bacterial straines were isolated from activated sludge and compost The theoretical part contains a description of PHA, extremophilic bacteria and detection techniques for the determination of polyhydroxyalkanoates. In the experimental part, the presence of the phaC gene, which is crucial for the ability to produce PHA, was first determined by PCR and bacterial strains were also subjected to DNA sequencing of 16S rRNA gene which enabled preliminary taxonomical classification of the isolates. In the next part of the experimental work, the conditions for biomass growth and PHA production were optimized. Suitable carbon source, the ideal temperature for cultivation and the influence of precursors on the production of copolymers were studied and identified. The composition and proportion of PHA were determined spectrophotometrically and by GC-FID. Finally, visual screening of PHA accumulation inside bacterial cells was performed using fluorescence microscopy.
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Analýza bakteriálních izolátů a DNA ze společenstva ze čtyř různých sukcesních ploch na území po těžbě hnědého uhlí
Chroňáková, Alica ; Halbritter, A. ; Krištůfek, Václav ; Biró, B.
Microbial communities of 4 different stages (initial [0 years after heaping of the excavated material], early [10], middle [20] and late [42] of primary succession process were studied. They were characterized by genotyping methods: ARDRA - Amplified ribosomal DNA restriction analyses and box-PCR - genomic fingerprinting using specific primer for boxA-element. Box-PCR method was chosen for comparison of methodological approaches used for characterization of heterotrophic bacterial communities. ARDRA patterns of eubacterial communities were so complex and there weren´t shown any differences among individual communities on different stages of primary succession process. Actinomycetes communities were analyzed using two sets of primers: Actinomycetes 1. (243F-1378R, Heuer et al. 1997) and Actinomycetes 2. (243F-A3R; Monciardini et al. 2002). Fingerprints, which were obtained using both of actinomycetes specific primers, showed differences between communities on the early and later stages of primary succession process. Boy-PCR analysis is still in data processing and will be published later
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