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Biotechnological production of yeast metabolites under combined stress
Baťková, Zuzana ; Byrtusová, Dana (referee) ; Dzurendová, Simona (advisor)
This master's thesis focuses on utilizing waste whey for the biotechnological production of carotenogenic yeast biomass and its valuable metabolites such as carotenoids, ubiquinone, ergosterol, fatty acids, and -glucans. Combined stress conditions were used to evaluate the influence on the yeast biomass metabolites production. These metabolites find applications in various industries, for example pharmaceutical, cosmetic, and food. The thesis is divided into theoretical and experimental parts. The theoretical section describes the composition of whey and its valorization in various industries. Selected carotenogenic yeasts and their metabolites are also presented. Furthermore, the impact of nutritional, oxidative stress, and the influence of metals on the production of yeast biomass and metabolites are described. The experimental part is reporting the whey treatment, yeasts biomass production and methods for the determination of individual metabolites. Five strains of red yeasts were used in this thesis: Cystofilobasidium macerans (CCY 10-1-2), Phaffia rhodozyma (CCY 77-1-1), Sporobolomyces shibatanus (CCY 19-9-6), Rhodotorula kratochvilovae (CCY 20-2-26) and Rhodotorula toruloides (CYY 62-2-4). The strain Cystofilobasidium macerans showed the best results for the utilization of whey substrate under the influence of combined stress, especially in biomass, lipid, and -glucan production. A promising strain in carotenoid production under combined stress was Sporobolomyces shibatanus.
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Characterization of biological effects of extracts obtained from yeasts and autotrophic microorganisms
Špačková, Dominika ; Bendová, Agáta (referee) ; Szotkowski, Martin (advisor)
Carotenogenic yeasts and microscopic autotrophic microorganisms are increasingly being studied in the biotechnology industry because of their ability to produce a variety of substances with a wide range of utilization. These substances mainly include lipids and carotenoids, which are used in the food and pharmaceutical industries. The aim of this bachelor thesis is to determine the way these microorganisms behave in the presence of stress factors, and which of them would have a positive effect on the production of metabolites that can be further used in various industries. The theoretical part is dedicated to a characterization of substances produced by carotenogenic yeasts, cyanobacteria and microalgae, which will be further analyzed in the experimental part. The metabolites mainly include carotenoids, which are a wide group of naturally occurring pigments that possess, among other things, antioxidant and provitamin activity. No less important substances produced by these organisms are lipids, which are part of cell membranes, they also serve as a reservoir of energy, and the substances contained in lipids are part of the precursors of many hormones. Yeast cell walls also contain -glucans, which have many beneficial effects on immunity and human health. Due to this fact, interest in their research have been rising recently. The analytical methods used to determine these substances are also mentioned, as well as stress factors and their effects on the cells of the mentioned organisms. The experimental part deals with the exposure of these microorganisms to oxidative stress and stress caused by heavy metals. The aim was to analyze which concentrations of these stress factors are lethal to microorganisms, and which of them are optimal for increase of the production of the above-mentioned substances. Of the yeast strains, the strains Sporidiobolus pararoseus and Rhodotorula mucilaginosa achieved increased production of metabolites due to selenate ions. In contrast, cobalt had a beneficial effect only on the accumulation of lipids and -glucans in cells. The positive effect of selenium was also observed in the strain Chlorella minutissima and Arthrospira maxima, however, the most remarkable changes in the production of metabolites were achieved by the addition of the optimal concentration of the solution inducing oxidative stress.
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Processsing of food supplements based on medical types of mushrooms
Vetchá, Vendula ; Uhlířová, Renata (referee) ; Hoová, Julie (advisor)
This diploma thesis deals with the characterization of nutrition substances in medical mushrooms, specifically Ganoderma lucidum and Hericium erinaceus. It also focuses on the biological impact of these rare mushrooms on human cell lines HaCaT and Caco-2, and investigates the production of pro-inflammatory cytokines IL-6 and IL-8 by immortalized keratinocytes and colon cancer cells after exposure to aqueous extracts prepared from mushroom powder. The theoretical part of the thesis deals with the characterization of mushrooms in terms of morphological characteristics and their taxonomic classification. It further studies the effects of edible and medicinal species and provides a detailed description of medicinal mushrooms such as Ganoderma lucidum, Lentinula edodes, Flammulina velutipes, Grifola frondosa, Hericium erinaceus and Pleurotus ostreatus, thesis informing about their effects, utilization and possibilities for application in the food and pharmaceutical industries. It mentions possible negative effects of mushrooms on consumers' health and presents methods for processing mushrooms and their use in the food industry. Finally, it focuses on the possibilities of in vitro toxicity testing and provides a brief overview of commonly used cytotoxicity tests such as the MTT test and LDH test. The experimental part of the thesis deals with the preparation of water extracts from mushroom powders and their characterization in terms of nutritional and biologically active substances. HPLC analysis is used to monitor the content of lipophilic substances and vitamin C in mushroom samples. To confirm the biological effect on human health, antimicrobial tests were performed using four model microorganisms, and cytotoxicity tests were performed on two human cell lines. Lastly, a sensory analysis of instant vegetable soup enriched with mushroom powder was performed. Biological effects of the analyzed powdered extracts from Ganoderma lucidum mushroom have been confirmed, especially their cytotoxic effect on colon epithelial tumor cells. However, the application of reishi in the food industry is not easy due to the presence of triterpenoids that cause a bitter taste. On the other hand, the safety of using the Hericium erinaceus mushroom in food has been confirmed through cytotoxicity tests. Sensory analysis results have shown a positive effect of Hericium erinaceus on the taste profile of instant vegetable soups.
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Optimization of methods for the isolation and analysis of beta-glucans produced by carotenogenic yeasts
Šimková, Eva ; Márová, Ivana (referee) ; Szotkowski, Martin (advisor)
The aim of this thesis was to optimize the methods of isolation and analysis of -glucan polysaccharides produced by carotenogenic yeasts. As part of the experiment, the yeast Rhodotorula toruloides, which was cultivated in laboratory bioreactors, was selected as the production strain. Theoretical part describes the subject of carotenogenic yeasts and the metabolites they produce. The main metabolite that this work deals with is the polysaccharide -glucan. -glucans are polysaccharides that form the cell wall in the yeast cell. They are known for their immunostimulating effects, antioxidant effects, promote wound healing and help lower cholesterol and blood glucose levels. The experimental part of the work deals with the cultivation of yeast biomass and its characterization. Furthermore, optimization of the enzymatic determination of -glucans using a commercial enzymatic kit and isolation of -glucans from yeast biomass. As part of the experiment, yeast biomass extraction methods were also further developed, enabling the separation and analysis of -glucans, lipids, lipidic substances and proteins from a small sample of yeast biomass in one extraction procedure.
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Characterization of biological effects of extracts obtained from yeasts and autotrophic microorganisms
Špačková, Dominika ; Bendová, Agáta (referee) ; Szotkowski, Martin (advisor)
Carotenogenic yeasts and microscopic autotrophic microorganisms are increasingly being studied in the biotechnology industry because of their ability to produce a variety of substances with a wide range of utilization. These substances mainly include lipids and carotenoids, which are used in the food and pharmaceutical industries. The aim of this bachelor thesis is to determine the way these microorganisms behave in the presence of stress factors, and which of them would have a positive effect on the production of metabolites that can be further used in various industries. The theoretical part is dedicated to a characterization of substances produced by carotenogenic yeasts, cyanobacteria and microalgae, which will be further analyzed in the experimental part. The metabolites mainly include carotenoids, which are a wide group of naturally occurring pigments that possess, among other things, antioxidant and provitamin activity. No less important substances produced by these organisms are lipids, which are part of cell membranes, they also serve as a reservoir of energy, and the substances contained in lipids are part of the precursors of many hormones. Yeast cell walls also contain -glucans, which have many beneficial effects on immunity and human health. Due to this fact, interest in their research have been rising recently. The analytical methods used to determine these substances are also mentioned, as well as stress factors and their effects on the cells of the mentioned organisms. The experimental part deals with the exposure of these microorganisms to oxidative stress and stress caused by heavy metals. The aim was to analyze which concentrations of these stress factors are lethal to microorganisms, and which of them are optimal for increase of the production of the above-mentioned substances. Of the yeast strains, the strains Sporidiobolus pararoseus and Rhodotorula mucilaginosa achieved increased production of metabolites due to selenate ions. In contrast, cobalt had a beneficial effect only on the accumulation of lipids and -glucans in cells. The positive effect of selenium was also observed in the strain Chlorella minutissima and Arthrospira maxima, however, the most remarkable changes in the production of metabolites were achieved by the addition of the optimal concentration of the solution inducing oxidative stress.
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Malotechnology of application forms herbal remedies and bee products
HÝBL, Marian
The aim of this thesis is to clarify the concept of malotechnology, focusing on herbal preparations, bee products and build three functional herbal preparations enriched with bee products. Herbs and bee products have always been people very treasured commodity. They played an irreplaceable role in healing but also in the production of alcohol, perfumes, teas and conventional foods. Now, these commodities are valued for their versatility in the pharmaceutical, cosmetic, chemical and food industries. The main objective of this work is to propose three different herbal preparations enriched with bee products. Description of the chemical composition, effects, application possibilities and contraindications. There have been proposed and described in detail following preparations: 1) Tincture of sage enriched with propolis, high quality with maximum content of active substances exhibiting strong antimicrobial and immunomodulatory effects 2) Oat meal rich in -glucans with honey, enriched with royal jelly. Nutritionally and dietetic valuable, tasty porridge with a high content of -glucans, with a recommended daily dose of royal jelly and honey in a biologically active state. 3) Hemp-propolis body cream enriched with sea buckthorn oil and marigold. Natural body cream enriched with extracts of the highest quality from cannabis sativa, propolis, sea buckthorn and marigold with versatility use (healing of burns and various damage and chronic skin diseases).
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