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Preparation and properties of cyanide-transforming enzymes and their analytical uses
Šťastná, Katarína ; Martínková, Ludmila (advisor) ; Čermáková, Michaela (referee)
Cyanides are compounds that are both naturally occurring and used in various industries. Their negative effects on human health, but also on the environment, have been demonstrated and their occurrence should therefore be monitored. The aim of this master thesis is the enzymatic determination of free cyanide, focusing on the characterization and application of specific enzymes within enzymatic cascades. The main objectives include the production and purification of key enzymes involved in cyanide conversion, namely cyanide hydratase (CynH), formamidase (AmiF), and cyanide dihydratase (CynD). The enzymes' catalytic properties and effectiveness in detecting cyanide were assessed. Spectrophotometric techniques were used to measure intermediates and products enzymatic reactions. The development, testing and optimization of enzyme cascades was carried out to demonstrate their feasibility and efficiency in cyanide determination. As a result, two selective, sensitive and rapid enzymatic assays for the detection of free cyanide have been developed. The proposed assays require small amounts of samples, enzymes and other components and enable rapid cyanide determination in less than two hours. Overall, this research contributes to the advancement of methods for cyanide analysis, with potential applications...
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