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Characterization of individual phases of cultivation of red yeasts
Valentová, Lucie ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
This bachelor thesis deals with the problematic of metabolic activity and morphology of several biotechnologically significant red yeasts - Rhodotorula glutinis, Sporobolomyces shibatanus and Phaffia rhodozyma during the cultivation time. These yeasts were cultivated on glucose medium, during the cultivation samples were taken for the analysis of produced metabolites. The amount of -carotene, astaxanthin, coenzyme Q and ergosterol were monitored. The amount of biosynthesised lipids and fatty acid profile was also investigated. HPLC/PDA and GC/FID were used for the analysis of these metabolites. Yeasts were cultivated 56 hours (they reached early stationary phase). The highest production of carotenoid pigments showed Phaffia rhodozyma (1856.58 g/g), 42 % of this amount represents -carotene. The highest lipids biosynthesis efficiency was reached by Rhodotorula glutinis. To study the inner structure of yeast cells the Fluorescence Life-time Imaging was used, the Autofluorescence of carotenoids and fluorescence of lipid bodies induced by lipophilic dye Nile red was observed. This method is very useful for imaging cell inner structure (membranes, some kinds of organelles). The changes that the cell undergoes during its evolution could be quantified too. The relationship between the level of carotenoid concentration, membrane rigidity and metabolic shift was found.
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Use of carotenogenic yeasts to production of lipid soluble metabolites
Mariničová, Veronika ; Hlaváček, Viliam (referee) ; Márová, Ivana (advisor)
Carotenoids are fat-soluble chemical compounds that occur as natural pigments in many plants and protect them from sunlight. Lipids are also essential lipophilic substances and they are part of biomembranes. Their main function is primarily to serve as a power supply for the cell, protective function and thermal protection against adverse environmental influences. This bachelor thesis deals with cultivation of selected carotenoid yeast genes, subsequent isolation of carotenoids and other lipid substances, which can be used as a source of potentially beneficial substances for the pharmaceutical or cosmetic industry. The theoretical part deals with the description of carotenoid yeasts, chemical composition and biosynthesis of the metabolites produced, and description of the methods used for their determination. The experimental part is focused mainly on the production of lipid substances by various strains of yeasts using cheap waste substrates and the application of exogenous stress (nutritional stress) to the biotechnological overproduction of selected metabolites using the modification of the production medium. The content of carotenoids, ergosterol and coenzyme Q was analyzed by high performance liquid chromatography with a PDA detector. The lipids accumulated in yeast cells were determined by gas chromatograph with a FID detector. In this work the strains of Sporobolomyces pararoseus, Sporobolomyces metaroseus, Rhodotorula glutinis and Cystofilobasidium infirmominiatum were studied. As waste substrates and carbon sources were used glycerol, which is produced as a waste product in the production of biofuels and whey as an unusable product in dairy technology. The best production on waste substrates was observed in the strains Rhodotorula glutinis and Cystofilobasidium infirmominiatum.
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Comparison of growth and productivity of autotrophic algae and red yeasts
Kouřilová, Xenie ; Rapta, Marek (referee) ; Márová, Ivana (advisor)
The submitted bachelor thesis focuses on the comparison of growth and productivity of red yeasts Rhodotorula aurantiaca and Xanthophyllomyces dendrorhous and autotrophic algae Chlamydomonas reinhardtii and Chlorella vulgaris under different physical conditions. The theoretical part contains the characteristics of selected micro-organisms, stress factors caused by temperature, radiation, combination of aeration and radiation and osmotic stress. Furthermore, knowledge on metabolites and the possibilities of their analysis are mentioned. Physical stresses, namely 2%, 3%, 4% and 5% solution of sodium chloride, intense radiation, absence of lighting, 8°C and 30°C, were applied to selected strains in cultivation. Under these conditions, changes in production of carotenoids, fatty acids, ergosterol, ubiquinone and proteins were being observed. A significant inhibitive effect on all the above mentioned micro-organisms was brought about by the use of 5% solution NaCl. The highest production of metabolites with X. dendrorhous was discovered when 4% solution of NaCl was used. R. aurantiaca demonstrated higher substance utilization at 30°C. For the selected algae strains were the used stresses lethal. Within the cultivation process, the possibilities of co-cultivation of yeasts and algae were also tested, which will be the subject of further studies.
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A comparison of metabolic activity and use of microalgae and pigment-formed yeasts
Szotkowski, Martin ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
Carotenoids are natural pigments occurring in plants and many microorganisms, such as algae, yeast and bacteria. They represent the most common group of antioxidants with significant biological effect. Lipids are an essential component of all living organisms. They are the source and the reservoir of energy for organisms, which can also be used in the petrochemical industry for the production of biofuels. This thesis deals with the production properties of carotenoids and lipids by selected strains of yeasts, algae, and canobacteria, in order to apply the acquired knowledge and find cheap suitable alternatives for microbial biotechnological production of these metabolites using waste substrates. Carotenoids, coenzyme Q, and ergosterol in cells were determined by liquid chromatography. The lipid content and fatty acid profile was analyzed by gas chromatography. Cell morphology and localization of selected metabolites were analyzed by fluorescence lifetime imaging microscopy. In this work yeast genera Sporobolomyces, Cystofilobasidium and Rhodotorula were used. As the representatives of the algae and cyanobacteria strains of Desmodesmus, Cyanothece, Chlamydomonas, Synechoccocus and Chlorella strains were studied. In the overall comparison, the yeast strains were more productive than algae and cyanobacteria. The highest carotenoid production was found in S. pararoseus, C. capitatum and R. mucilaginosa cells, while the highest lipid yield was observed in strains of C. infirmominiatum and S. metaroseus.
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Determination of residual substances from the preparation of colloidal systems based on biopolymers
Borůvková, Tereza ; Vávrová, Milada (referee) ; Mravec, Filip (advisor)
Volatile organic compounds could be isolated and concentrated from liquid solution using methods, which are based on analysis of equilibrium gas phase, because of their volatility. The aim of this bachelor´s thesis is determination the amount of residual tert-butanol, belonging to volatile organic compounds, in the samples after lyophilisation. Samples were prepared by mixing hydrofobic sonde and hydrofilic hyaluronan. For mixing these components it is necessary to add organic solvent tert-butanol, which ensures stable environment. Thanks to its presence mixing of probe and hyaluronan occurs. Consecutive lyopfilisation of the sample leads to the disruption of a water cover of hyaluronan and nonpolar hydrogen atoms, which are hidden in it, can interact with hydrofobic probe. Bonding the probe is important in oncology, where this fact could be used in targeted distribution of pharmaceuticals. However presence of tert-butanol is unacceptable in the final product, mainly in medical applications. The object this study is the minimization of residual tert-butanol amount in final lyophilised cake. The method of equilibrium gas phase analysis (static Head-Space) by gas chromatography with flame ionization detection (GC/FID) was used for the determination of its content.
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