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Biodegradability of bacterial natural and modified polyesters and their composites
Pala, Martin ; Babák, Libor (referee) ; Obruča, Stanislav (advisor)
Presented work was focused on biodegradability of bacterial natural and modified polyesters and their composites. The first part of the work was focused on study of influence of PHA granules structure on their biodegradability using selected enzymes and influence of physiological conditions on PHA stability. Overall, tested polymer either in crytalinne or amorphous phase seems to be rezistent to attack of seleced hydrolytic enzymes such as lipases or proteases and is stable in simulated physiological fluids as well. Because of thies results, it is possible to use tested PHA materials in biomedical applications requiring rather resistant biomaterials. Second part of the work was focused on microbial degradation of modified PHA materials considering their potential environmental impact. Both mixed thermophillic culture originaly used in wastewater treatment plant and bacterial strain Delftia acidovorans were employed for biodegradation tests. Composites containing chlorine PHB and PHB films modified using plasticizers were tested. Films containing chlorine PHB cause inhibition of biomass growth to both tested cultures. The highest rate of degradation (31%) was observed in presence of bacterial culture with film containing 10% chlorine PHB. The results show that used microbial population is important factor affecting biodegradability.
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Production of polyhydroxyalkanoates from waste substrates and their isolation
Grossová, Marie ; Babák, Libor (referee) ; Márová, Ivana (advisor)
The aim of this work is to study the possibility of microbial production of polyhydroxyalkanoates (PHA). PHA can be used as biodegradable materials. Bacterial strain Cupriavidus necator was used for laboratory production of PHA. This bacterium was cultivated in medium with various precursors to produce copolymers of 3HB with 3HV or 4HB. Another part of the work was aimed at cultivation of C. necator on different waste substrates, especially oils, with the aim to achieve the highest production of polymer. Another large part of the thesis is dedicated to isolation strategies of PHA using enzymes. Commercially used proteases – alcalase and pancreatin – can be used with advantages for digestion of bacterial cells. A number of optimization experiments showed that application of proteases leads to enhancement of PHA purity to about 13%. Purity increase up to 90 % was achieved by adding a surfactant, which promotes the solubility of non-PHA forming polymer. This surfactant increases the purity of 20 % when compared to control. The last part of presented work deals with the use of enzyme solution isolated from Bacillus subtilis medium. Its application to C. necator culture led to the yield of polymer at a purity exceeding 95 %. These results could represent the basis for new isolation strategies, which can lead to more efficient yield of PHA.
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Regulation of polyhydroxybutyrate production in Wautersia eutropha
Grossová, Marie ; Čarnecká, Martina (referee) ; Márová, Ivana (advisor)
The aim of this work is study of microbial production of biodegradable polymers (polyhydroxyalkanoates; polyhydroxybutyrate PHB). Bacterium Wautersia eutropha was used for laboratory production of PHB. In this bacterium application of exogenous stress factors (osmotic shock, hydrogen peroxide and ethanol) was tested to enhance PHB production. Influence of exogenous stress on bacterial culture depended on stress factor concentration and time of stress application. Substantial influence on PHB yield exhibited changes of biomass production during stress experiments. The main part of this work is focused on study of PHB production in microbial cells exposed to stress in different growth phases. Application of strong stress in early phases of growth led to PHB and biomass decrease. The most suitable seems to be application of stress at the beginning of stationary phase accompanied by increase of PHB formation. Concentration of stress factor could be optimized too. The highest yields were obtained after addition of 0.5 % ethanol into production medium at 60th hour (PHB increase 21 %). These results can form a basis for new fermentation strategy which leads to enhanced effectiveness of PHB production.
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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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Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) employing selected members of the genus Aeromonas
Čeparová, Klára ; Slaninová, Eva (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis deals with the study of polyhydroxyalkanoates (PHA) production by Aeromonas spp. Four bacterial strains were selected for this study: Aeromonas aquatica (DSM 100827), Aeromonas bivalvium (DSM 19111), Aeromonas hydrophila (CCM 7770) and Aeromonas cavernicola (CCM 7641). The presence of phaC gene responsible for PHA synthesis was verified by polymerase chain reaction (PCR). In three of four selected strains (A. aquatica, A. bivalvium, A. hydrophila) the presence of phaC gene was confirmed. Strains A. cavernicola did not contain the phaC gene. PHA synthesis on different carbon substrates was further tested on strains with confirmed presence of the phaC gene. The aim of the thesis was to optimize the cultivation conditions of bacterial cultures for the PHA synthesis. The production of polyhydroxybutyrate (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) P(3HB-co-3HHx) was monitored. The evaluation of polymer presence and concentration in biomass was determined by gas chromatography. For cultivation, decanoic acid, caproic acid, octanoid acid, glukose, sucrose, fructose were used as carbonaceous substrates. The highest of both PHB and the copolymer P(3HB-co-3HHx) were recorded by A. aquatica with addition of decanoid acid at a concentration of 1 g/l to the production medium. Among the sugar substrates, cultivation on sucrose was the most successful with, resulting in the highest biomass concentration.
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Biodegradability of bacterial natural and modified polyesters and their composites
Pala, Martin ; Babák, Libor (referee) ; Obruča, Stanislav (advisor)
Presented work was focused on biodegradability of bacterial natural and modified polyesters and their composites. The first part of the work was focused on study of influence of PHA granules structure on their biodegradability using selected enzymes and influence of physiological conditions on PHA stability. Overall, tested polymer either in crytalinne or amorphous phase seems to be rezistent to attack of seleced hydrolytic enzymes such as lipases or proteases and is stable in simulated physiological fluids as well. Because of thies results, it is possible to use tested PHA materials in biomedical applications requiring rather resistant biomaterials. Second part of the work was focused on microbial degradation of modified PHA materials considering their potential environmental impact. Both mixed thermophillic culture originaly used in wastewater treatment plant and bacterial strain Delftia acidovorans were employed for biodegradation tests. Composites containing chlorine PHB and PHB films modified using plasticizers were tested. Films containing chlorine PHB cause inhibition of biomass growth to both tested cultures. The highest rate of degradation (31%) was observed in presence of bacterial culture with film containing 10% chlorine PHB. The results show that used microbial population is important factor affecting biodegradability.
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Production of polyhydroxyalkanoates from waste substrates and their isolation
Grossová, Marie ; Babák, Libor (referee) ; Márová, Ivana (advisor)
The aim of this work is to study the possibility of microbial production of polyhydroxyalkanoates (PHA). PHA can be used as biodegradable materials. Bacterial strain Cupriavidus necator was used for laboratory production of PHA. This bacterium was cultivated in medium with various precursors to produce copolymers of 3HB with 3HV or 4HB. Another part of the work was aimed at cultivation of C. necator on different waste substrates, especially oils, with the aim to achieve the highest production of polymer. Another large part of the thesis is dedicated to isolation strategies of PHA using enzymes. Commercially used proteases – alcalase and pancreatin – can be used with advantages for digestion of bacterial cells. A number of optimization experiments showed that application of proteases leads to enhancement of PHA purity to about 13%. Purity increase up to 90 % was achieved by adding a surfactant, which promotes the solubility of non-PHA forming polymer. This surfactant increases the purity of 20 % when compared to control. The last part of presented work deals with the use of enzyme solution isolated from Bacillus subtilis medium. Its application to C. necator culture led to the yield of polymer at a purity exceeding 95 %. These results could represent the basis for new isolation strategies, which can lead to more efficient yield of PHA.
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|
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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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Regulation of polyhydroxybutyrate production in Wautersia eutropha
Grossová, Marie ; Čarnecká, Martina (referee) ; Márová, Ivana (advisor)
The aim of this work is study of microbial production of biodegradable polymers (polyhydroxyalkanoates; polyhydroxybutyrate PHB). Bacterium Wautersia eutropha was used for laboratory production of PHB. In this bacterium application of exogenous stress factors (osmotic shock, hydrogen peroxide and ethanol) was tested to enhance PHB production. Influence of exogenous stress on bacterial culture depended on stress factor concentration and time of stress application. Substantial influence on PHB yield exhibited changes of biomass production during stress experiments. The main part of this work is focused on study of PHB production in microbial cells exposed to stress in different growth phases. Application of strong stress in early phases of growth led to PHB and biomass decrease. The most suitable seems to be application of stress at the beginning of stationary phase accompanied by increase of PHB formation. Concentration of stress factor could be optimized too. The highest yields were obtained after addition of 0.5 % ethanol into production medium at 60th hour (PHB increase 21 %). These results can form a basis for new fermentation strategy which leads to enhanced effectiveness of PHB production.
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