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Production of mcl-PHA by selected bacteria
Eremka, Libor ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The aim of this work is to study the possibility of microbial production of polyhydroxyalkanoates with medium chain length (mcl-PHA). For production two bacterial strains were used: Pseudomonas putida and Pseudomonas aeruginosa. Theoretical part of bachelor thesis deals with the biosynthesis of polyhydroxyalkanoates, their material properties and microbial producers. In experimental part selected bacteria were cultivated on different substrates to evaluate their ability to utilize various carbon sources and accumulate mcl-PHA. Content and composition of mcl-PHA accumulated in biomass were analyzed by gas chromatography with FID. Pseudomonas putida produced more mcl-PHA in medium containing canola oil as sole carbon source when compared with Pseudomonas aeruginosa. P. putida produced 2,11% mcl-PHA in medium containing glycerol as carbon source.
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Influence of selected parameters on biotechnological production of polyhydroxyalkanoates
Eremka, Libor ; Pekař, Miloslav (referee) ; Obruča, Stanislav (advisor)
The aim of this work is to study microbial production of polyhydroxyalkanoates (PHA). Theoretical part is focused on production of PHA using microoganisms and transgenic plants. Bacterial strain Cupriavidus necator H16 was used for laboratory production of PHA. Various waste oils were used as sole carbon and energy source. Salt of propionic adic and 1-propanol were used as intermediate for 3-hydroxyvalerate monomer (3HV) unit. Incorporation of 3HV to polymer can improve material features of PHA. The major part of experimental work was focused to study influence of aeration (concentration of dissolved oxygen) to bacterial growth, selected metabolic pathways and formation of PHA. Furthermore, influence of aeration to monomer composition of polymer was evaluated. According to experimental conclusion of this work it was approved dependance between aeration and monomer composition of PHA. Moreover, it was approved that higher concentration of oxygen supports bacterial growth and influences PHA content in cells. In addition, NADPH is one of the substrates influencing flux of acetyl-CoA throughout the metabolism; higher intracellular concentration of NADPH inhibits TCA cycle and enhances accumulation of PHA in cells. For this reason, specific enzymatic aktivity of several selected intracelular enzymes were measured, including those enzymes which can generate NADPH.
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Influence of selected parameters on biotechnological production of polyhydroxyalkanoates
Eremka, Libor ; Pekař, Miloslav (referee) ; Obruča, Stanislav (advisor)
The aim of this work is to study microbial production of polyhydroxyalkanoates (PHA). Theoretical part is focused on production of PHA using microoganisms and transgenic plants. Bacterial strain Cupriavidus necator H16 was used for laboratory production of PHA. Various waste oils were used as sole carbon and energy source. Salt of propionic adic and 1-propanol were used as intermediate for 3-hydroxyvalerate monomer (3HV) unit. Incorporation of 3HV to polymer can improve material features of PHA. The major part of experimental work was focused to study influence of aeration (concentration of dissolved oxygen) to bacterial growth, selected metabolic pathways and formation of PHA. Furthermore, influence of aeration to monomer composition of polymer was evaluated. According to experimental conclusion of this work it was approved dependance between aeration and monomer composition of PHA. Moreover, it was approved that higher concentration of oxygen supports bacterial growth and influences PHA content in cells. In addition, NADPH is one of the substrates influencing flux of acetyl-CoA throughout the metabolism; higher intracellular concentration of NADPH inhibits TCA cycle and enhances accumulation of PHA in cells. For this reason, specific enzymatic aktivity of several selected intracelular enzymes were measured, including those enzymes which can generate NADPH.
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Production of mcl-PHA by selected bacteria
Eremka, Libor ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The aim of this work is to study the possibility of microbial production of polyhydroxyalkanoates with medium chain length (mcl-PHA). For production two bacterial strains were used: Pseudomonas putida and Pseudomonas aeruginosa. Theoretical part of bachelor thesis deals with the biosynthesis of polyhydroxyalkanoates, their material properties and microbial producers. In experimental part selected bacteria were cultivated on different substrates to evaluate their ability to utilize various carbon sources and accumulate mcl-PHA. Content and composition of mcl-PHA accumulated in biomass were analyzed by gas chromatography with FID. Pseudomonas putida produced more mcl-PHA in medium containing canola oil as sole carbon source when compared with Pseudomonas aeruginosa. P. putida produced 2,11% mcl-PHA in medium containing glycerol as carbon source.
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