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Electrochemical evolution of hydrogen peroxide for biotechnological applications
Runštuková, Nikola ; Kratochvíl, Matouš (referee) ; Ehlich, Jiří (advisor)
This master‘s thesis deals with the connection of two distinct fields, electrochemistry and biotechnology, where electrochemically generated peroxide was subsequently used in bacterial cultivation. The peroxide was synthesized in an electrolytic cell by reducing oxygen at the cathode to which suitable electrical potential was applied. The aim of this thesis was to investigate the dependence of the amount of hydrogen peroxide produced on a few selected parameters, such as cathode potential, electrode material, the geometry of the measuring cell, or the stirring speed, and then to apply these findings in the continuous stressing of the bacterium Cupriavidus necator H16, in order to increase the production of polyhydroxyalkanoates. The selected parameters were first tested in small-scale measuring cells and then experiments were carried out on a larger scale, in Erlenmeyer flasks. During the experiments, the amount of electric current produced was measured and also the peroxide concentration was examined spectrophotometrically along with the oxygen concentration measured by using an optical sensor. Based on the obtained results, bacteria were cultivated in flasks with a built-in system of steel electrodes, due to which oxidative stress in the form of in situ generated hydrogen peroxide was continuously applied. The bacterial cultures were evaluated by measuring the change in optical density, gravimetric analysis and GC-FID analysis. It was found that even small amounts of hydrogen peroxide (~25 M) can induce a stress response in the organism in the form of increased polyhydroxybutyrate (PHB) synthesis. A percentage increase in the PHB content of the biomass was achieved, however, the overall polymer yield was lower due to lower biomass growth.

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