Original title: Isolace a charakterisace katechol 1,2-dioxygenasy kvasinky Candida tropicalis
Translated title: Isolation and characterization of catechol 1,2-dioxygenase of Candida tropicalis
Authors: Jechová, Jana ; Stiborová, Marie (advisor) ; Martínková, Markéta (referee)
Document type: Master’s theses
Year: 2011
Language: cze
Abstract: [cze] [eng]
Kvasinka Candida tropicalis je mikroorganismus vykazující vysokou toleranci vůči fenolu a silnou schopnost jej degradovat. Tato kvasinka je schopná využít fenol jako zdroj uhlíku a energie, aniž by produkovala sekundární polutant. Z kvasinky C. tropicalis byla isolována katechol-1,2-dioxygenasa. K purifikaci bylo využito chromatografie na DEAE Sepharose a gelové permeační chromatografie na Sephadexu G-100. Uvedeným postupem byl získán homogenní enzym. Enzymová aktivita byla sledována pomocí HPLC (úbytek katecholu a nárůst cis,cis-mukonátu). Na základě měření aktivity při různých teplotách bylo určeno teplotní optimum, jeho hodnota je 30řC. Kinetické charakteristiky byly studovány při různém pH. Hodnota Km a Vmax byla 0,52 mM a 17,2 nM/min pro úbytek katecholu a 0,34 mM a 12,6 nM/min pro nárůst cis,cis-mukonové kyseliny při optimálním pH reakce, pH 7,6. Candida tropicalis yeast is a microorganism that possesses high tolerance for phenol and strong phenol degrading activity. This yeast is capable of utilizing phenol as the sole source of carbon and energy without formation of any secondary waste product. Catechol-1,2- dioxygenase was isolated from cytosolic fraction of this yeast by the procedure consisting of chromatography on DEAE-Sepharose and gel permeation chromatography on Sephadex G- 100. The catechol-1,2-dioxygenase was purified to homogeneity. The enzyme activity was followed by HPLC (catechol consumption and/or cis,cis-muconic acid formation). The activity profiles at different temperatures showed temperature optimum of 30řC. Kinetic characterizations were studying in different values of pH. The values of Km and Vmax of 0,52 mM and 17,2 nM/min for consumption of catechol, respectively, and 0,34 mM and 12,6 nM/min for formation of cis,cis-muconic acid, respectively, were found at optimum pH of the reaction, pH 7,6.
Keywords: biodegradation; Candida tropicalis; catechol-1,2-dioxygenase; phenol; biodegradace; Candida tropicalis; fenol; katechol-1,2-dioxygenasa

Institution: Charles University Faculties (theses) (web)
Document availability information: Available in the Charles University Digital Repository.
Original record: http://hdl.handle.net/20.500.11956/34540

Permalink: http://www.nusl.cz/ntk/nusl-477317


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Universities and colleges > Public universities > Charles University > Charles University Faculties (theses)
Academic theses (ETDs) > Master’s theses