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Application of chemical and physical stresses in the late phase of growth to selected strains of microorganisms
Langer, Marek ; Holub, Jiří (referee) ; Szotkowski, Martin (advisor)
A wide spectrum of microorganisms is used in the biotechnological industry for production of meaningful metabolites; including carotenoids and chlorofyles with antioxidant effects, fatty acids, ubiquinone and ergosterol. Ways to increase yield of metabolites to maximum capacity are being researched to increase efficiency and economical sustainability in industry. One of the ways is the application of chemical stress factors on microorganisms as is described in this thesis. In the theoritical part certain microorganisms, their important intracellular metabolites and their metabolism are established. The last subchapter covers the usage of stress factors in industry. In the experimental part each microorganism was subjected to various concentrations of sodium chloride, cobalt(II) sulfate and iron(III) chloride. Significant increase of lipid production was registered after an application of sodium chloride on yeast strain Rhodosporium toruloides. An increase in production of torularhodin in Sporidiobolus pararoseus occured after application of iron salt. After an application of sodium chloride the growth of torularhodin in strains Cystofilobasidium macerans a Rhodosporidium toruloides was significant. Sodium chloride was also a suitable stress factor for lycopene formation in Rhodotorula mucilaginosa. The most meaningful stress factor of algae was cobalt(II) sulfate which increased metabolite production in Scenedesmus obliquus. Cyanobacteria Anabaena torulosa also attained a significant production in the presence of sodium chloride.
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Influence of stress and nutritional conditions on carotenogenic yeast and microalgal metabolism
Sikorová, Pavlína ; Byrtusová, Dana (referee) ; Szotkowski, Martin (advisor)
This bachelor thesis describes the infuence of biological stress (co-cultivations) on the growth and metabolism of selected carotenogenic yeasts and microalgae. The metabolites of these microorganisms are chlorophyll and carotenoids, which are natural pigments and antioxidants. In theory, co-cultivation is a cultivation where microorganisms mutually support each other in the growth and production of metabolites. This would cause the increased production of previously mentioned chlorophyll and carotenoids. The theoretical part of the work is focused on the description of selected species of yeasts and microalgae and further discusses in more detail, the topics of stress factors depending on the growth and metabolism of microorganisms. The experimental part then deals with different types of cultivation and cocultivation and tries to optimize the production media and find the best symbiotic yeasts and microalgae. Furthemore the issue of different ratios of microorganisms in co-cultivations is also addressed here. The cultivated yeasts strains were Rhodotorula kratochvilovae, Rhodosporidium toruloides and Phaffia rhodozyma. And microalgae strains were Desmodesmus acutus, Desmodesmus quadricauda, Coccomyxa sp., Chlorella sorokiniana, Chlamydomonas reinhardtii and Scenedesmus obliquus. Cultivated cyanobacterium was Synechoccocus nidulans. The most successful co-cultivation experiment was the one with R. kratochvilovae and yeast Desm. quadricauda. This experiment was very succesful in all aspects.
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Preparation of microbial, plant and algal extracts to use in cosmetics for infants
Janderová, Šárka ; Dzurická, Lucia (referee) ; Márová, Ivana (advisor)
This bachelor thesis deals with the preparation of a cosmetic product for infants with the addition of active substances from cyanobacteria, microalgae, or plants. In the theoretical part, a review was made, which not only summarizes the basic information about these organisms, but also the antioxidants contained in these microorganisms. In the practical part, several types of extracts from cyanobacteria, algae and plants were prepared and characterized. The content of phenolic substances, chlorophylls and content of carotenoids were measured spectrophotometrically. Content of soluble proteins was determined by Hartree-Lowry method and the fatty acid profile in biomass was obtained by GC. Furthermore, the SPF factor was measured using the spectrophotometric method the Mansur equation. Antioxidant activity was determined by using the TEAC method. Finally, two of the best extracts were selected for the preparation of cosmetic emulsions. The sensory analysis of prepared emulsions was performed for the period of two weeks.
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Biological effects of complex extracts from yeast and algae biomass
Bočán, David ; Byrtusová, Dana (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis was focused on optimalization of extractions of active compounds from microalgae and carotenogenic yeasts. Other part of this work was focused on characterization of these extracts and their tests of cytotoxicity on human keratinocytes. In theoretical part a literary research which summarizes the basic properties and cultivation conditions of used microorganisms was conducted. Then there were listed an examples of antioxidant compounds found in these microorganisms as well as their properties. Finally the term cytotoxicity and methods of its measurement were clarified. The practical part of this thesis consists of series of extractions from mentioned microorganisms. These extracts were characterized by the content determination of phenolic and flavonoid compounds and also chlorophyll and carotenoid compounds. Determination of antioxidant capacity and SPF were done too. Lastly the cytotoxicity of chosen extracts was measured using the MTT test.
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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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Cultivation of carotenogenic yeasts in the presence of biological stress induced by selected strains of autotrophic microalgae
Sikorová, Pavlína ; Byrtusová, Dana (referee) ; Szotkowski, Martin (advisor)
Presented Master’s thesis was focused on biological and nutrient stress in cultivations of green microalgae and carotenogenic yeasts. The focus of interest was production of biomass in different types of cultivation media, production of metabolites (mainly carotenoids and chlorophylls) and also production of lipids and lipophilic substances. All types of aplied stress was used to lead to an increased production of biomass and metabolites. The theoretical part deals with introduction of individual genera of carotenogenic yeast and green microalgae. The types of cultivation stress were also described. In addition, the analytic methods for qualitative and quantitative analysis are also introduced. The experimental part was focused on the study of biomass, carotenoids, chlorophylls, coenzym Q, sterols and lipids production. Biological stress was induced by cocultivation of microalgae and yeasts together.Nutrient stress meant adding macroelements or waste oils to the medium. Yeasts of interest were Rhodosporidium toruloides, Rhodotorula kratochvilovae and Sporobolomyces pararoseus. Microalgae was represented by Desmodesmus acutus, Desmodesmus armatus, Scenedesmus obliquus, Desmodesmus velitaris, Desmodesmus communis, Coccomyxa sp. Chlamydomonas reinhardtii and Chlorella minutissima. Generally the most successful multicultivator experiment was cocultivation with yeast R. kratochvilovae. In the case of cultivation on waste oils, the best producers of biomass and metabolites were co-cultivation experiments with the yeast R. toruloides. In microalgal experiments, it was found that nutritional stress in the form of glycerol added to the medium had an inhibitory effect on the growth and metabolism of microalgae.
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Development of fortified gluten-free cereals for specific nutrition
Dohnalová, Radka ; Fialová, Lenka (referee) ; Márová, Ivana (advisor)
The bachelor thesis was focused on the development of enriched gluten-free cereals for special nutrition. The fortified cereal product included processed microbial biomass, either as a freeze-dried powder or as an extract, for which the phenolic content, antioxidant content and protein content were determined. The theoretical part includes findings from professional literature on gluten-free cereals, pseudocereals and their chemical composition. Subsequently, a review on microalgae and yeasts was prepared including a list of their metabolites. The literature search focused on green microalgae and carotenogenic yeasts. In the experimental part, the characterization of the selected pseudocereals and optimization of the prepared extracts of the selected microalgae and carotenogenic yeast were firstly carried out. The selected pseudocereals were amaranth, buckwheat and quinoa. The green microalgae strain Chlamydomonas reinhardtii and the carotenogenic yeast strain Cystofilobasidium macerans were selected for enrichment of the pseudocereals. Carbohydrates, proteins, fatty acids, -glucans, gluten, vitamins, phenolic substances, flavonoids and antioxidants were determined in the pseudocereals. Total fatty acids, lipophilic substances, phenolic substances and total antioxidant activity were detected in the microbial biomass. Finally, mixed cereal products were prepared from pseudocereal flour, microbial biomass and microbial extract. The cytotoxicity of the selected extracts was determined by MTT assay. It was found that the addition of algal biomass alone or algal extract to cereal flours resulted in a significant increase in protein. The yeast ethanol extract increased the phenolic and antioxidant content of the cereal products.
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Production of enriched mixed biomass by co-cultivation of selected strains of yeast and microalgae
Langer, Marek ; Němcová, Andrea (referee) ; Szotkowski, Martin (advisor)
The use of yeast and microalgae in industry is constantly increasing. These microorganisms are able to produce significant amounts of lipids, which can be used, for example, for the production of biodiesel. Like their other metabolites, such as carotenoids, chlorophylls, ubiquinone and ergosterol, they can be used in the food industry. Since these chemical substances also have biological effects on the human organism, they are used in food supplements, but are also being investigated as possible medicines. The presented work deals with the possibilities of making the cultivation of these microorganisms cheaper. One of these options is co-cultivation, or mixed cultivation, which aims to increase biomass and metabolite yields. Another investigated possibility is the replacement of financially expensive substrates for the nutrition of microorganisms with cheap waste from the food industry. The algae Scenedesmus ecornis, Desmodesmus communis, Chlamydomonas reinhardtii and Coccomyxa sp. were used in the experimental work. The yeast strains used were Rhodotorula kratochvilovae and Rhodotorula toruloides. Added waste products were oils (hemp, frying and coffee), molasses, whey and waste glycerol. Analysis of the obtained biomass revealed that the highest lipid yields were achieved when coffee and frying oil were used. The experiment was completed by pilot cultivation in a bioreactor, to which cultures of Scenedesmus ecornis and Rhodotorula kratochvilovae were added. Coffee oil and glycerol were added as a source of nutrients.
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Evaluation of the physiological status of European beech in its plantings on localities following the gradient of its ecological valence
Palouš, Daniel ; Albrechtová, Jana (advisor) ; Tomášková, Ivana (referee)
European beech (Fagus sylvatica L.) is one of the dominant tree species in Europe, presenting a keystone species for forest ecosystem functioning. The ongoing climate change alters the environmental conditions in the areas of the current distribution of European beech, leading to stress. Concerning the critical health status of forests in the Czech Republic and other European countries, it's even more urgent to monitor the health status of forests. For this, tools for early detection of a stand's physiological status decline are needed. The aforementioned tools can consist of indicators of the physiological status suggested by this thesis. This thesis aims to evaluate the applicability and sensitivity of the preselected set of indicators of the physiological status of European beech to the gradient of environmental conditions (temperature, precipitation) within its ecological valence. Four stands of European beech presenting the gradient of beech's ecological valence were observed over the peak of one vegetation season. Stand Kocanda presented the microclimatic and ecological optimum; stand Hradecko was on the lower margin of the altitudinal gradient of European beech distribution and presented the limit of low water availability; stand Deštné was on the upper margin of the altitudinal gradient of...
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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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