Original title: Biotransformace fenolických látek enzymovými systémy kvasinky Candida tropicalis a bakterie Comamonas testosteroni
Translated title: Biotransformation of phenols by enzymatic systems of Candida tropicalis yeast and Comamonas testosteroni bacteria
Authors: Vilímková, Lenka ; Stiborová, Marie (advisor) ; Entlicher, Gustav (referee) ; Mareš, Jaroslav (referee)
Document type: Doctoral theses
Year: 2011
Language: cze
Abstract: [cze] [eng]
Kvasinka Candida tropicalis a bakterie Comamonas testosteroni se vyznačují schopností využívat fenol jako jediný zdroj uhlíku a energie. V naší laboratoři se zabýváme zejména studiem cytoplasmatických enzymů zodpovědných za první a druhý krok odbourání fenolu: NADPH-dependentní fenolhydroxylasy C. tropicalis i C. testosteroni a katechol 1,2- dioxygenasy C. tropicalis. Cílem naší práce je izolovat a částečně charakterizovat tyto enzymy. Fenolhydroxylasa C. tropicalis byla purifikována použitím následující kombinace metod: chromatografie a rechromatografie cytosolárních proteinů na DEAE Sepharose, frakční precipitace polyethylenglykolem 6000 a gelové permeační chromatografie na Sephacrylu S-300. Extracelulární fenolhydroxylasa C. testosteroni byla získána frakční precipitací kultivačního média polyethylenglykolem 6000 a gelovou permeační chromatografií na 4B Sepharose a Sephacrylu S-300. Katecholdioxygenasa byla z cytosolu C. tropicalis izolována za použití metod: chromatografie a rechromatografie cytosolárních proteinů na DEAE Sepharose, lyofilizace enzymového preparátu a gelové permeační chromatografie na Sephadexu G-100 Enzymová aktivita byla měřena buď pomocí HPLC nebo spektrofotometricky. Čistota enzymů byla sledována elektroforézou SDS-PAGE. Molekulová hmotnost enzymů byla určena pomocí... Candida tropicalis yeast and bacteria Comamonas testosteroni have been considered to be able to metabolize phenol and utilize it as the only source of carbon and energy. In our laboratory we investigated the cytoplasmic enzymes responsible for the first and second step of phenol degradation, NADPH-dependent phenol hydroxylase of both C. tropicalis and C. testosteroni and catechol 1,2-dioxygenase of C. tropicalis. The aim of our study was to isolate and partially characterize those enzymes. Phenol hydroxylase purification consisted of preparation of cytosol from C. tropicalis yeast by fraction centrifugation, chromatography and re-chromatography on a column of DEAE Sepharose, fractionation by precipitation of the enzyme with polyethylene glycol 6000 and gel permeation chromatography on a column of Sephacryl S-300. Extracellular phenol hydroxylase of C. testosteroni was purified by fraction precipitation with polyethylene glycol 6000 and by gel permeation chromatography on 4B Sepharose and Sephacryl S-300. Catechol 1,2-dioxygenase was purified using the procedure consisting of: chromatography and re- chromatography on a column of DEAE Sepharose, lyophilization of the enzyme and gel permeation chromatography on a column of Sephadex G-100. The enzyme activity was determined by two methods: use of HPLC...
Keywords: Biotransformation; Candida tropicalis; Comamonas testosteroni; enzymes; oxidation; phenols; Biotransformace; Candida tropicalis; Comamonas testosteroni; enzymy; fenolické látky; oxidace

Institution: Charles University Faculties (theses) (web)
Document availability information: Available in the Charles University Digital Repository.
Original record: http://hdl.handle.net/20.500.11956/47270

Permalink: http://www.nusl.cz/ntk/nusl-311550


The record appears in these collections:
Universities and colleges > Public universities > Charles University > Charles University Faculties (theses)
Academic theses (ETDs) > Doctoral theses