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New fungal fucosidases and their synthetic potential
Glozlová, Michaela ; Beránek, Martin (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michaela Glozlová Supervisor: prof. PharmDr. Martin Beránek, Ph.D. Consultants: doc. RNDr. Pavla Bojarová, Ph.D., Mgr. Pavlína Nekvasilová Title of diploma thesis: New fungal fucosidases and their synthetic potential The literary research focuses on α-L-fucosidases, their occurrence, functions, and use in the synthesis of fucosylated compounds, which were also a part of the research, particularly their presence in the human body and involvement in pathological conditions. Furthermore, it explores the oligosaccharides found in human milk, their impact on infant health, and their synthesis, which can be facilitated through the use of α-L-fucosidases. The experimental section involves the analysis of a library of filamentous fungi with the aim of identifying a representative strain with α-L-fucosidase activity towards pNP-α-L-fucoside. Recombinant α-L-fucosidases were selected based on searches in genetic databases and prepared. The feasibility of their production in Pichia pastoris and Escherichia coli was tested, and the produced enzymes were purified by ion exchange chromatography, characterized and their regioselectivity and synthetic potential for the synthesis of fucosylated...
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New fungal fucosidases and their synthetic potential
Glozlová, Michaela ; Beránek, Martin (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michaela Glozlová Supervisor: prof. PharmDr. Martin Beránek, Ph.D. Consultants: doc. RNDr. Pavla Bojarová, Ph.D., Mgr. Pavlína Nekvasilová Title of diploma thesis: New fungal fucosidases and their synthetic potential The literary research focuses on α-L-fucosidases, their occurrence, functions, and use in the synthesis of fucosylated compounds, which were also a part of the research, particularly their presence in the human body and involvement in pathological conditions. Furthermore, it explores the oligosaccharides found in human milk, their impact on infant health, and their synthesis, which can be facilitated through the use of α-L-fucosidases. The experimental section involves the analysis of a library of filamentous fungi with the aim of identifying a representative strain with α-L-fucosidase activity towards pNP-α-L-fucoside. Recombinant α-L-fucosidases were selected based on searches in genetic databases and prepared. The feasibility of their production in Pichia pastoris and Escherichia coli was tested, and the produced enzymes were purified by ion exchange chromatography, characterized and their regioselectivity and synthetic potential for the synthesis of fucosylated...
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Enzymatic detoxification of cyanide using cyanide hydratases
Sedova, Anastasia ; Rucká, Lenka ; Glozlová, Michaela ; Novotný, Petr ; Martínková, Ludmila ; Bojarová, Pavla
Cyanide, a strong chelator of metals in vital enzymes and proteins, is very toxic for most living organisms. Wastewaters from the mining, metallurgical and chemical industries contain significant concentrations of free cyanide. Though it can be largely eliminated by physicochemical methods, these processes may in turn lead to the formation of other waste. In addition, no effective methods have been found to neutralize cyanide spills coming from industrial accidents. The use of enzymes to remove cyanide is an attractive alternative, which is environmentally friendly and applicable to high cyanide concentrations. Cyanide hydratases (CynHs) are of considerable interest for the decomposition of hazardous cyanide wastes. Here we investigate the biochemical properties of new CynHs from Stereum hirsutum and Exidia glandulosa, it is of fundamental importance to preserve the enzyme activity at alkaline pH as cyanide waste is stored under these conditions.
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