|
|
Employment of bacterium Tepidimonas taiwanensis for production of polyhydroxyalkanoates on waste substrates
Vidláková, Michaela ; Pernicová, Iva (referee) ; Obruča, Stanislav (advisor)
This diploma thesis is focused on screening PHA production using thermophilic bacterium Tepidimonas taiwanensis and on the study of possible use of grape pomace, molasses and waste paper as a cheap carbon substrate for culturing the characterized bacteria. At first, testing of basic cultivation parameters was performed, including carbon substrate concentration, oxygen availability, ability to utilize nitrogen sources and selected disaccharides. PHA production from waste substrates was tested in three ways. In the first, pre-prepared solids-free hydrolysates from raw materials were used as the carbon source. The second and third procedures were performed by dosing waste materials directly into the mineral media, which differed only in the presence or absence of the enzyme preparation enabling release of fermentable sugars. The most intensive increase in culture and the highest production of PHA was recorded on grape hydrolyzate. The biomass concentration in this sample reached up to 4.8 g/L with a content of 59 % PHA. On the other hand, the addition of grape marc directly to the production medium did not work at all, which was probably due to the presence of a large amount of inhibitory substances from the pomace. The situation was similar with molasses and waste paper, where the bacterium Tepidimonas taiwanensis was able to grow and possibly produce PHA only to a small extent. The work also managed to characterize the effect of temperature and pH on the activity of the enzyme cocktail Viscozyme L and to determine the temperature and pH optimum PHA synthase of the bacterial strain Tepidimonas taiwanensis in the cell lysate.
|
|
|
Utilization of selected extremophilic bacteria for bioplastic production
Vidláková, Michaela ; Obručová,, Hana (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis is focused on the study of the biotechnological potential of thermophilic bacteria Tepidiphilus thermophilus, Tepidimonas taiwanensis and Tepidimonas fonticaldi. The aim of this work was to investigate the ability of bacteria to produce polyhydroxyalkanoates (PHA). The theoretical part includes the characteristics of PHA and extremophilic bacteria. In the experimental part, the investigated bacteria were tested by molecular technique PCR for the presence of the phaC gene encoding for the first class PHA synthase. The presence of the phaC gene was confirmed in bacterial strains Tepidiphilus thermophilus and Tepidimonas taiwanensis. The ability of PHA production on various carbon sources was examined for all the selected microorganisms, optimal temperatures for cultivation were determined, and the ability to synthesize copolymers was investigated by adding suitable precursors to the production medium. The most intensive PHA production was recorded in the bacterial strain Tepidimonas taiwanensis, but other strains were also able to produce PHA. The ability to produce P(3HB-co-3HV) or P(3HB-co-4HB) copolymers has also been demonstrated in some strains. A significant amount of P(3HB-co-3HV) was obtained using the Tepidimonas taiwanensis strain. Morever, Tepidimonas taiwanensis strain was also tested for its ability to use various model lignocellulose hydrolysates to form PHA. The highest PHA gain was obtained using model hydrolysate of softwood. Furthermore, the ability of proteases production by selected bacterial strains was tested. The presence of these enzymes was detected in the Tepidimonas taiwanensis strain, which was followed by determination of the temperature optima of the proteases, the pH optima of the proteases and also of their molecular weights.
|
|
|
Employment of bacterium Tepidimonas taiwanensis for production of polyhydroxyalkanoates on waste substrates
Vidláková, Michaela ; Pernicová, Iva (referee) ; Obruča, Stanislav (advisor)
This diploma thesis is focused on screening PHA production using thermophilic bacterium Tepidimonas taiwanensis and on the study of possible use of grape pomace, molasses and waste paper as a cheap carbon substrate for culturing the characterized bacteria. At first, testing of basic cultivation parameters was performed, including carbon substrate concentration, oxygen availability, ability to utilize nitrogen sources and selected disaccharides. PHA production from waste substrates was tested in three ways. In the first, pre-prepared solids-free hydrolysates from raw materials were used as the carbon source. The second and third procedures were performed by dosing waste materials directly into the mineral media, which differed only in the presence or absence of the enzyme preparation enabling release of fermentable sugars. The most intensive increase in culture and the highest production of PHA was recorded on grape hydrolyzate. The biomass concentration in this sample reached up to 4.8 g/L with a content of 59 % PHA. On the other hand, the addition of grape marc directly to the production medium did not work at all, which was probably due to the presence of a large amount of inhibitory substances from the pomace. The situation was similar with molasses and waste paper, where the bacterium Tepidimonas taiwanensis was able to grow and possibly produce PHA only to a small extent. The work also managed to characterize the effect of temperature and pH on the activity of the enzyme cocktail Viscozyme L and to determine the temperature and pH optimum PHA synthase of the bacterial strain Tepidimonas taiwanensis in the cell lysate.
|
|
|
Utilization of selected extremophilic bacteria for bioplastic production
Vidláková, Michaela ; Obručová,, Hana (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis is focused on the study of the biotechnological potential of thermophilic bacteria Tepidiphilus thermophilus, Tepidimonas taiwanensis and Tepidimonas fonticaldi. The aim of this work was to investigate the ability of bacteria to produce polyhydroxyalkanoates (PHA). The theoretical part includes the characteristics of PHA and extremophilic bacteria. In the experimental part, the investigated bacteria were tested by molecular technique PCR for the presence of the phaC gene encoding for the first class PHA synthase. The presence of the phaC gene was confirmed in bacterial strains Tepidiphilus thermophilus and Tepidimonas taiwanensis. The ability of PHA production on various carbon sources was examined for all the selected microorganisms, optimal temperatures for cultivation were determined, and the ability to synthesize copolymers was investigated by adding suitable precursors to the production medium. The most intensive PHA production was recorded in the bacterial strain Tepidimonas taiwanensis, but other strains were also able to produce PHA. The ability to produce P(3HB-co-3HV) or P(3HB-co-4HB) copolymers has also been demonstrated in some strains. A significant amount of P(3HB-co-3HV) was obtained using the Tepidimonas taiwanensis strain. Morever, Tepidimonas taiwanensis strain was also tested for its ability to use various model lignocellulose hydrolysates to form PHA. The highest PHA gain was obtained using model hydrolysate of softwood. Furthermore, the ability of proteases production by selected bacterial strains was tested. The presence of these enzymes was detected in the Tepidimonas taiwanensis strain, which was followed by determination of the temperature optima of the proteases, the pH optima of the proteases and also of their molecular weights.
|
guest ::
RSS feed.