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Use of diffrent nutrition source by selected carotenogenic yeasts
Čačková, Katarína ; Starečková, Terezie (referee) ; Márová, Ivana (advisor)
Carotenoids are the most common naturally occurring terpenoid pigments. Because of their positive biological functions, carotenoids represent a group of important molecules for the pharmaceutical, chemical, food and feed industries. This work was realized as a comparative study focused on use of some waste substrates as nutritional sources for production of carotenoid pigments by red yeast Rhodotorula aurantiaca. Yeast were cultivated in different media with selected waste substrates – apple fiber, whey, wheat etc. To some media hydrolytic enzymes prepared from some fungal strains were added. Most of waste substrates seems to be useful for R. aurantiaca cultivation. As the best nutritional sources for enriched biomass and carotenoid production apple fiber or wheat processed by mixed preparative of extracellular hydrolases from Fusarium solani were found.
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Use of Waste Substrates to Production of Enriched Yeast Biomass
Starečková, Terezie ; Demnerová, Kateřina (referee) ; Vávrová, Milada (referee) ; doc.PharmDr.Petr Babula, Ph.D. (referee) ; Márová, Ivana (advisor)
Yeasts are like other organisms constantly exposed to environmental influences. Their survival depends on the skills to adapt to environmental changes, including the ability to use various alternative sources of nutrients. In presented PhD thesis carotenogenic yeast belonging to the genera Rhodotorula, Sporobolomyces and Cystofilobasidium were tested for ability to use of selected waste substrates, and also subjected to several types of exogenous stress effects and mutations in order to increase the production of microbial biomass enriched with specific metabolites. As alternative nutrient sources derived from waste substrates from agricultural and farm production apple peel, pulp, corn germ and more were tested. Yeasts were also exposed to osmotic, oxidative and combined stress (benefits of various concentrations of NaCl and H2O2 to the culture media), followed by metal ions of selenium and chromium in concentrations of 0.01 mM, 0.1 mM and 1 mM. The effect of mutagen methanesulfonic acid ethyl ester was tested too. In all experiments the adaptivity of cells, morphological changes, color pigments produced by the media while some important fungal metabolites production and changes in chromosomal DNA fragmentation were analyzed. In order to evaluate potential changes in the yeast genome after treatment with mutagen and stress factors methods for isolation of intact chromosomal DNA and DNA analysis by pulsed field gel electrophoresis was optimized. The amount of produced metabolites was mainly analyzed by RP-HPLC with UV/VIS and MS detection. The work has been shown that most strains are able to use waste substrates and produced selected target metabolites. Biomass, for example, in R. aurantiaca on apple fiber was about 7 g/l and in C. capitatum cultivated on modified whey reached to 9 g/l. Amount of produced carotenoids by R. aurantiaca cultivated on wheat germ and maize after enzymatic hydrolysis by F. solani was 1.01 mg/g and S. roseus on pasta 4.3 mg/g. The values of ergosterol synthesis in R. aurantiaca are on the apple shells around 4.8 mg/g, in S. roseus on pasta with the enzymatic hydrolysis of P. chrysosporium 8.9 mg/g. The best substrate for biomass production and induction of carotenoids are waste substartes containing a mixture of simple and complex carbohydrates enriched with the addition of nitrogen compounds. Potential cytotoxic effect of stress factors of low concentrations was demonstrated. Red yeast genome was able to distribute by optimized PFGE, the karyotype of tested yeasts contain 11 or more chromosomes with visible differences between yeast species and genera. During exchange internship the ability of recombinant yeast S. cerevisiae to convert xylose to xylitol, which would be achieved by increasing the production of bioethanol as alternative fuel sources was studied. It turned out that both ligninocellulose materials to bioethanol production, as well as various waste substrates for microbial synthesis of carotenoids would reduce costs for industrial production of yeast metabolites, as well as to reduce the negative burden on the environment.
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Analysis of biologically active substances in some cereal products
Valentová, Radka ; Starečková, Terezie (referee) ; Márová, Ivana (advisor)
Presented bachelor thesis is focused on study of biologically active substances in cereal products. In theoretical part over view of cereal active substances, predominantly phenolic compounds were introduced. In practical part 15 kinds of cereal products were analyzed. In these materials some group parameters - total phenolics, total flavonoids and total antioxidant activity were measured by spectrophotometry. RP-HPLC/UV-VIS method was performed for analysis of selected individual flavonids. The highest content of phenolic compounds was measured in cereals containing fruits (raspberries, blueberries, tropical and red fruit), cacao, crusty fruits (hazel nuts, sunflower seeds) and buckwheat. All cereal products contain catechine, epicatechine, catechin gallate, epicatechin gallate, chlorogenic and ferulic acid. As a part of bachelor thesis simple sensory analysis was performed and consumer questionnaires were evaluated.
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Use of Waste Substrates to Production of Enriched Yeast Biomass
Starečková, Terezie ; Demnerová, Kateřina (referee) ; Vávrová, Milada (referee) ; doc.PharmDr.Petr Babula, Ph.D. (referee) ; Márová, Ivana (advisor)
Yeasts are like other organisms constantly exposed to environmental influences. Their survival depends on the skills to adapt to environmental changes, including the ability to use various alternative sources of nutrients. In presented PhD thesis carotenogenic yeast belonging to the genera Rhodotorula, Sporobolomyces and Cystofilobasidium were tested for ability to use of selected waste substrates, and also subjected to several types of exogenous stress effects and mutations in order to increase the production of microbial biomass enriched with specific metabolites. As alternative nutrient sources derived from waste substrates from agricultural and farm production apple peel, pulp, corn germ and more were tested. Yeasts were also exposed to osmotic, oxidative and combined stress (benefits of various concentrations of NaCl and H2O2 to the culture media), followed by metal ions of selenium and chromium in concentrations of 0.01 mM, 0.1 mM and 1 mM. The effect of mutagen methanesulfonic acid ethyl ester was tested too. In all experiments the adaptivity of cells, morphological changes, color pigments produced by the media while some important fungal metabolites production and changes in chromosomal DNA fragmentation were analyzed. In order to evaluate potential changes in the yeast genome after treatment with mutagen and stress factors methods for isolation of intact chromosomal DNA and DNA analysis by pulsed field gel electrophoresis was optimized. The amount of produced metabolites was mainly analyzed by RP-HPLC with UV/VIS and MS detection. The work has been shown that most strains are able to use waste substrates and produced selected target metabolites. Biomass, for example, in R. aurantiaca on apple fiber was about 7 g/l and in C. capitatum cultivated on modified whey reached to 9 g/l. Amount of produced carotenoids by R. aurantiaca cultivated on wheat germ and maize after enzymatic hydrolysis by F. solani was 1.01 mg/g and S. roseus on pasta 4.3 mg/g. The values of ergosterol synthesis in R. aurantiaca are on the apple shells around 4.8 mg/g, in S. roseus on pasta with the enzymatic hydrolysis of P. chrysosporium 8.9 mg/g. The best substrate for biomass production and induction of carotenoids are waste substartes containing a mixture of simple and complex carbohydrates enriched with the addition of nitrogen compounds. Potential cytotoxic effect of stress factors of low concentrations was demonstrated. Red yeast genome was able to distribute by optimized PFGE, the karyotype of tested yeasts contain 11 or more chromosomes with visible differences between yeast species and genera. During exchange internship the ability of recombinant yeast S. cerevisiae to convert xylose to xylitol, which would be achieved by increasing the production of bioethanol as alternative fuel sources was studied. It turned out that both ligninocellulose materials to bioethanol production, as well as various waste substrates for microbial synthesis of carotenoids would reduce costs for industrial production of yeast metabolites, as well as to reduce the negative burden on the environment.
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Study of metabolic changes in carotenogenic yeasts cultivated under different conditions
Starečková, Terezie ; Breierová, Emília (referee) ; Márová, Ivana (advisor)
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.
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Use of diffrent nutrition source by selected carotenogenic yeasts
Čačková, Katarína ; Starečková, Terezie (referee) ; Márová, Ivana (advisor)
Carotenoids are the most common naturally occurring terpenoid pigments. Because of their positive biological functions, carotenoids represent a group of important molecules for the pharmaceutical, chemical, food and feed industries. This work was realized as a comparative study focused on use of some waste substrates as nutritional sources for production of carotenoid pigments by red yeast Rhodotorula aurantiaca. Yeast were cultivated in different media with selected waste substrates – apple fiber, whey, wheat etc. To some media hydrolytic enzymes prepared from some fungal strains were added. Most of waste substrates seems to be useful for R. aurantiaca cultivation. As the best nutritional sources for enriched biomass and carotenoid production apple fiber or wheat processed by mixed preparative of extracellular hydrolases from Fusarium solani were found.
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Analysis of biologically active substances in some cereal products
Valentová, Radka ; Starečková, Terezie (referee) ; Márová, Ivana (advisor)
Presented bachelor thesis is focused on study of biologically active substances in cereal products. In theoretical part over view of cereal active substances, predominantly phenolic compounds were introduced. In practical part 15 kinds of cereal products were analyzed. In these materials some group parameters - total phenolics, total flavonoids and total antioxidant activity were measured by spectrophotometry. RP-HPLC/UV-VIS method was performed for analysis of selected individual flavonids. The highest content of phenolic compounds was measured in cereals containing fruits (raspberries, blueberries, tropical and red fruit), cacao, crusty fruits (hazel nuts, sunflower seeds) and buckwheat. All cereal products contain catechine, epicatechine, catechin gallate, epicatechin gallate, chlorogenic and ferulic acid. As a part of bachelor thesis simple sensory analysis was performed and consumer questionnaires were evaluated.
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