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Study on PHA production in extremophiles from genus Bacillus and related genuses
Reinohová, Nikola ; Kouřilová, Xenie (referee) ; Pernicová, Iva (advisor)
This Bachelor thesis is focused on study of production od polyhydroxyalcanoteas by extremophilic bacteria of genus Bacillus and related genera. In this thesis were studied microorganisms from german and czech collections Ureibacillus composti DSM 171951, Alkalibacillus haloalkaliphilus DSM 5271, Alicyclobacillus acidocaldarius DSM 446, Halobacillus halophilus CCM 3527, Thermobacillus composti DSM 18247, Bacillus licheniformis CCM 2206 and isolated microorganisms from natural sources Aneurinibacillus thermoaerophilus LK7, Aneurinibacillus thermoaerophilus F109, Aneurinibacillus thermoaerophilus AFn2, Geobacillus thermodenitrificans F101, Geobacillus thermodenitrificans F102, Geobacillus stearothermophilus A12, Geobacillus sp. AH11. The theoretical part describes extremophilic microorganisms, polyhydroxyalkanoates and their applications. In the experimental part, the detection of PHA production at the genotype level was performed using the PCR method, where the presence of first and fourth class PHA synthases was detected. Detection of presence of the 16SrRNA gene was performed by PCR. PHA production was also tested at the phenotype level, where the ability of utilization of different carbon sources and the ability of microorganisms to form 4HB and 3HV copolymers with different precursors at different temperatures was monitored. Copolymers are very interesting because of their properties, which make them suitable for a wide range of applications. The microorganism Aneurinibacillus thermoaerophilus AFn2 proved to be the best representative for PHA production in this work, producing PHA up to 1,99 g/l and 3HV copolymer up to 0,49 g/l.
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Production, characterization and application of polymers of malic and glutamic acids
Čangelová, Katarína ; Kučera, Dan (referee) ; Obruča, Stanislav (advisor)
Subject of this bachelor thesis was study of biopolymers production using microorganisms. Polymer of malic acid was produced by Aureobasidium pullulans, a yeast-like fungus. Biosynthesis of polymer of glutamic acid was carried out by Bacillus licheniformis bacterium. Particular polymers, their microbial producers and possible application in different areas are characterised in the theoretical part of the thesis. Through experiments, the effect of various conditions on cell growth and production of these polymers was studied. Cultivation conditions were optimised in terms of polymers production. Subsequently, waste substrates - whey and poultry feathers were used. Microbially-produced polymer of malic acid was hydrolysed with 2 M sulfuric acid. Concentration of malic acid was analysed by IEC with a conductivity detector. Concentration of polymer of glutamic acid was determined by a spectrophotometric method using CTAB.
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Stanovení aktivity průmyslově významných enzymů produkovaných mikroorganismy izolovanými z potravinového odpadu
Lymarenko, Sofiia
The microbiome and the produced enzymes play an important role in processing food waste. Bacillus licheniformis is one of the representatives of microflora in composting and producer of industrially important hydrolytic enzymes: protease, lipase and cellulase. As part of the thesis, enzymatic activity using spectrophotometric methods was determined in seven strains of B. licheniformis for a period of 96 to 192 hours. In all seven strains, cellulase, protease and lipase activity was detected, which agrees with previously published literature. The highest cellulase activity determined by two methods was recorded for strain 3A-52 (1 U/ml) and 1A-52 (1.5 U/ml) after 24 hours of cultivation. The pH optimum for cellulase activity was 7, and the temperature optimum 50 °C to 60 °C. The maximum protease activity was detected after 120 and 144 hours of cultivation in two strains: 3A-52 (1.3 U/ml) and 3D-42 (1.4 U/ml). During the optimization, it was found that the pH optimum was 9 to 11 and the temperature optimum was 40 to 45 °C depending on the tested strain. The maximum lipase activity was approximately 0.7 U/ml after 144 hours of cultivation in 4D-41 and 3D-42. The temperature optimum was found at 50 to 60 °C and the range of pH optimum for this activity was very wide depending on the studied strain. For the strains with the highest enzymatic activity, a growth curve was determined, where the highest cellulase activity corresponded to the time of transition of the bacterial culture from the exponential phase to the stationary phase, and the highest lipase and protease activities were found later only at the transition from the stationary phase to the death phase. Strain 3A-52 and 3D-42 showed the highest activity for all three monitored enzymes. Subsequently, these strains isolated during composting can be used to produce enzymes industrially.
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Study on PHA production in extremophiles from genus Bacillus and related genuses
Reinohová, Nikola ; Kouřilová, Xenie (referee) ; Pernicová, Iva (advisor)
This Bachelor thesis is focused on study of production od polyhydroxyalcanoteas by extremophilic bacteria of genus Bacillus and related genera. In this thesis were studied microorganisms from german and czech collections Ureibacillus composti DSM 171951, Alkalibacillus haloalkaliphilus DSM 5271, Alicyclobacillus acidocaldarius DSM 446, Halobacillus halophilus CCM 3527, Thermobacillus composti DSM 18247, Bacillus licheniformis CCM 2206 and isolated microorganisms from natural sources Aneurinibacillus thermoaerophilus LK7, Aneurinibacillus thermoaerophilus F109, Aneurinibacillus thermoaerophilus AFn2, Geobacillus thermodenitrificans F101, Geobacillus thermodenitrificans F102, Geobacillus stearothermophilus A12, Geobacillus sp. AH11. The theoretical part describes extremophilic microorganisms, polyhydroxyalkanoates and their applications. In the experimental part, the detection of PHA production at the genotype level was performed using the PCR method, where the presence of first and fourth class PHA synthases was detected. Detection of presence of the 16SrRNA gene was performed by PCR. PHA production was also tested at the phenotype level, where the ability of utilization of different carbon sources and the ability of microorganisms to form 4HB and 3HV copolymers with different precursors at different temperatures was monitored. Copolymers are very interesting because of their properties, which make them suitable for a wide range of applications. The microorganism Aneurinibacillus thermoaerophilus AFn2 proved to be the best representative for PHA production in this work, producing PHA up to 1,99 g/l and 3HV copolymer up to 0,49 g/l.
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Production, characterization and application of polymers of malic and glutamic acids
Čangelová, Katarína ; Kučera, Dan (referee) ; Obruča, Stanislav (advisor)
Subject of this bachelor thesis was study of biopolymers production using microorganisms. Polymer of malic acid was produced by Aureobasidium pullulans, a yeast-like fungus. Biosynthesis of polymer of glutamic acid was carried out by Bacillus licheniformis bacterium. Particular polymers, their microbial producers and possible application in different areas are characterised in the theoretical part of the thesis. Through experiments, the effect of various conditions on cell growth and production of these polymers was studied. Cultivation conditions were optimised in terms of polymers production. Subsequently, waste substrates - whey and poultry feathers were used. Microbially-produced polymer of malic acid was hydrolysed with 2 M sulfuric acid. Concentration of malic acid was analysed by IEC with a conductivity detector. Concentration of polymer of glutamic acid was determined by a spectrophotometric method using CTAB.
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