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Biotechnological production of PHA employing Cupriavidus malaysiensis
Drábková, Kateřina ; Kovalčík, Adriána (referee) ; Obruča, Stanislav (advisor)
The subject of this diploma thesis is the study of the production of polyhydroxyalkanoates (PHA) employing Cupriavidus malaysiensis. The aim of this thesis is to obtain and characterize the polymer with the most advantageous properties. The theoretical part deals with polyhydroxyalkanoates and their biotechnological production. Then selected microorganisms producing 3-hydroxybutyrate and 4 hydroxybutyrate, P(3HB co 4HB), are described. After that, the theoretical part deals with the production of PHA employing Cupriavidus malaysiensis. The bacterial strains of Cupriavidus malaysiensis DSM 19416, 19379 and 25816 were used in the experimental part. The strain Cupriavidus necator H16 was used as a control strain. First, the ability to utilize various carbon sources by the given bacterial strains was determined. Then, the aim was to select the optimal carbon source, a precursor for the synthesis of a polymer with high content of 4-hydroxybutyrate, and to determine the ideal culturing conditions for the growth of the microorganisms, which have been studied, and the production of P(3HB co-4HB). A two-stage culture and bioreactor culture was also performed to produce the desired copolymer. Furthermore, the production of terpolymer of 3-hydroxybutyrate, 4 hydroxybutyrate and 3-hydroxyvalerate, P(3HB co 4HB co 3HV), was studied by single stage and two-stage cultivation. In the last part, the produced polymers were characterized by gas chromatography with flame ionization detector (GC-FID), size exclusion chromatography with multi-angle static light scattering (SEC-MALS), differential scanning calorimetry (DSC) and thermogravimetry.
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Biotechnological production of PHA employing Cupriavidus malaysiensis
Drábková, Kateřina ; Kovalčík, Adriána (referee) ; Obruča, Stanislav (advisor)
The subject of this diploma thesis is the study of the production of polyhydroxyalkanoates (PHA) employing Cupriavidus malaysiensis. The aim of this thesis is to obtain and characterize the polymer with the most advantageous properties. The theoretical part deals with polyhydroxyalkanoates and their biotechnological production. Then selected microorganisms producing 3-hydroxybutyrate and 4 hydroxybutyrate, P(3HB co 4HB), are described. After that, the theoretical part deals with the production of PHA employing Cupriavidus malaysiensis. The bacterial strains of Cupriavidus malaysiensis DSM 19416, 19379 and 25816 were used in the experimental part. The strain Cupriavidus necator H16 was used as a control strain. First, the ability to utilize various carbon sources by the given bacterial strains was determined. Then, the aim was to select the optimal carbon source, a precursor for the synthesis of a polymer with high content of 4-hydroxybutyrate, and to determine the ideal culturing conditions for the growth of the microorganisms, which have been studied, and the production of P(3HB co-4HB). A two-stage culture and bioreactor culture was also performed to produce the desired copolymer. Furthermore, the production of terpolymer of 3-hydroxybutyrate, 4 hydroxybutyrate and 3-hydroxyvalerate, P(3HB co 4HB co 3HV), was studied by single stage and two-stage cultivation. In the last part, the produced polymers were characterized by gas chromatography with flame ionization detector (GC-FID), size exclusion chromatography with multi-angle static light scattering (SEC-MALS), differential scanning calorimetry (DSC) and thermogravimetry.
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