The aim of this diploma thesis realized as a comparative study was the study of regulation of carotenoid and ergosterol production in several carotenogenic yeast strains. Yeasts were exposed to exogenous stress factors. Salt stress and oxidative stress (hydrogen peroxide) were reached by addition of NaCl and hydrogen peroxide into production media. Complex changes on metabolome (e.g. pigment and ergosterol production, RP-HPLC), proteome and genome were followed. Proteome changes were analyzed by PAGE-SDS and 2D electrophoresis. To isolation and analysis of chromosome DNA pulsed field gel electrophoresis (PFGE) was used. Six yeast strains were enrolled into the comparative study; three strains of the genus Rhodotorula and three strains of the genus Sporobolomyces. While yeasts Rhodotorula sp. were characterized by enhanced biomass as well as carotenoid production in normal and stress conditions, production of biomass by Sporobolomyces sp. was substantially lower. Carotenoid production in Sporobolomyces sp. was higher than in Rhodotorula sp.; the highest increase of was beta-carotene production was observed in Sporobolomyces salmonicolor cells stressed by salt (4x higher than in control) or peroxide (5x higher). Proteins were isolated from yeast cells by combination of mechanical and chemical disruption by glass beads and NaOh or SDS. Better yields were obtained by NaOH. Two staining methods were tested in PAGE-SDS protein analysis. Coomassie Brilliant Blue staining exhibited lower sensitivity, silver staining led to better visualisation of minor protein fractions too. 1D protein profiles was difficult to evaluate, therefore, 2D electrophoresis of selected strains (R.glutinis, R.rubra) was done. In yeast genome analysis by PFGE at minimum 7 DNA fractions were observed. These results probably are not final, further study will be needed for detailed characterization of red yeast genome.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:216333 |
| Date | January 2008 |
| Creators | Starečková, Terezie |
| Contributors | Breierová, Emília, Márová, Ivana |
| Publisher | Vysoké učení technické v Brně. Fakulta chemická |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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