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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 selected complex vitamin supplements
Bandarenka, Andrei ; Fialová, Lenka (referee) ; Márová, Ivana (advisor)
Modern society places great emphasis on a healthy lifestyle and balanced diet, leading to increased interest in products with high levels of vitamins, minerals, and other health-promoting substances. Consequently, there has been an increased interest in various dietary supplements, including so-called "greens" products. These supplements are designed to provide a concentrated source of nutrients from various green plants while also containing a high proportion of vitamins, minerals, antioxidants, and other bioactive compounds. The aim of this thesis is to characterize complex food supplements in terms of their composition and content of active ingredients to better understand their mechanisms of action and identify substances responsible for their health-promoting effects. The theoretical part of the work focuses on describing legislation of dietary supplements, including their characteristics and the evolution of interest in them over recent decades. Furthermore, the theoretical part addresses the description of basic components commonly found in supplements, such as vitamins, plants, algae, prebiotics, and probiotics. Finally, methods for determining these components and the most frequently used techniques for their analysis are described. In the experimental part, 12 different plant-based dietary supplements falling into the category known as "greens" were selected. The content of phenolic compounds, chlorophylls, carotenoids, and antioxidants were determined in these products. Additionally, the influence of these products on the growth of bacterial cultures and the metabolic activity of gram-negative and gram-positive bacteria was examined. Analysis of the vitamin C and B-group vitamins content in these supplements was also conducted. The test results showed that the examined supplements are a rich source of phenolic compounds, chlorophyll, antioxidants, and vitamin C. Furthermore, it was found that these products can be used as prebiotics in the diet. Based on the conducted tests, it was concluded that "greens" food supplements offer a wide range of health benefits and can be considered a useful contribution to a balanced diet and a healthy lifestyle. Practically, in all conducted tests, the products with a high matcha content achieved the best results, making this ingredient a key component for creating quality dietary supplements from plant-based ingredients.
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Bioactive compounds in cruciferous vegetables
KOVAČIKOVÁ, Karolína
This diploma thesis deals with bioactive compounds in cruciferous vegetables (Brassicaceae). The thesis aimed to determine the content of phenolic compounds, chlorophyll A, chlorophyll A+B, carotenoids and also vitamin C, in selected species of cruciferous vegetables: Brussel sprouts (Brassica oleracea var. gemmifera), Fodder Kale (Brassica oleracea var. acephala), Curly Kale (Brassica oleracea var. sabellica) and Ornamental Cabbage (Brassica oleracea var. capitata). The highest amount of chlorophyll A was represented in the sample of ornamental red cabbage, variety Pigeon Victoria F1. The highest content of chlorophyll B, chlorophyll A+B, carotenoids and also vitamin C was determined in curly green kale, a variety Tuscan. The highest amount of phenolic compounds was measured in curly red kale, variety Scarlet.
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Isolation and characterization of beta-glucans of natural origin
Ratsilouskaya, Lizaveta ; Hoová, Julie (referee) ; Márová, Ivana (advisor)
The bachelor thesis deals with the characterization of natural beta-glucans and other active substances isolated from macroalgae and yeasts. The used macroalgae samples differ in the pigment, origin and method of algae processing. The theoretical part focuses on seaweed and yeast species, beta-glucans, their sources and biological effects, other biologically active substances (such as polyphenols and antioxidants) and finally on methods for determining these biologically active substances. In the practical part, samples of selected macroalgae were analyzed, namely samples of Arame, Nori, Kombu, Wakame, Hijiki and Spaghetti. Aqueous, ethanol and acetone extracts of these algae were prepared in this work. The aqueous extracts were used for spectrophotometric determination of phenolic substances, flavonoids and antioxidant activity by the ABTS method. Furthermore, ethanol and acetone extracts of said macroalgae were used to determine total chlorophyll and carotenoid content spectrophotometric. The work also focused on beta-glucan content, analyzing macroalgae and yeast R. toruloides. Arame and Nori algae and R. toruloides were high in beta-glucans. However, beta-glucans were very low in other macroalgae.
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Production of lipids and lipid-soluble compounds by some yeasts and algae
Šimanský, Samuel ; Rapta, Marek (referee) ; Márová, Ivana (advisor)
The submitted bachelor thesis focuses on production of lipids and lipid-soluble compounds by selected strains of carotenogenic yeasts, autotrophic algae and cyanobacteria. Different forms of stress were applied on selected strains and its influence was subsequently compared. The theoretical part deals with the processing of knowledge about selected strains of yeast, algae and cyanobacteria. There are also mentioned the selected metabolites (lipids, carotenoids, ergosterol, ubiquinone and chlorophyll) and the possibilities of their determination. The experimental part deals primarily with the influence of nutritional stress on the production of selected metabolites and the yeast biomass, namely their growth properties on waste fat. Furthermore, there was monitored the effect of elevated and reduced temperature on the cultivation of yeasts, algae and cyanobacteria. Osmotic stress experiments were also carried out by the addition of NaCl. As the most suitable strain for decomposition of the waste fat was selected Sporidiobolus pararoseus, particularly because of its high content of carotenoids, ergosterols and ubiquinone in biomass. Changes in temperature, or increased osmotic pressure had in the most cases a negative effect on biomass production.
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Controlled production of lipids and lipidic substances by selected yeasts and microalgae
Szotkowski, Martin ; Kráčmar, Stanislav (referee) ; Huelva, Ines Garbayo Nores, University (referee) ; Márová, Ivana (advisor)
Karotenoidy jsou přírodní pigmenty vyskytující se v mikroorganismech jako jsou řasy, kvasinky a sinice. Představují nejrozšířenější skupinu antioxidantů s významným biologickým účinkem. V současnosti vzrůstá zájem o karotenoidy vzhledem k jejich příznivým vlivům na lidské zdraví. Chlorofyly jsou zelená fotosyntetická barviva, která nacházejí uplatnění v potravinářství jako intenzivní zelená barviva. Koenzym Q je znám svým pozitivním vlivem pro správnou funkci řady orgánů v lidském těle. Ergosterol je nedílnou součástí membrán kvasinek a hub. Je to provitamin D2, který je důležitou součástí imunitního systému. Mikrobiální lipidy, nebo také ‚‚Single cell oils‘‘ jsou charakteristické vysokým obsahem zdraví prospěšných nenasycených mastných kyselin, které lze využít ve farmacii či kosmetice. Mikrobiální lipidy jsou dále studovány jako alternativa pro výrobu biopaliv. Dizertační práce byla zaměřena na studium a možnosti optimalizace produkce lipidů a lipidických látek vybranými kmeny karotenogenních kvasinek, mikrořas a sinic. V rámci práce byly testované kvasinky rodu Rhodotorula, Rhodosporidium, Cystofilobasidium a Sporidiobolus podrobené kultivacím na sérií médií s různými C/N poměry v rozsahu 13 až 100, obsahujících upravené odpadní substráty z potravinářského průmyslu. Vybrané kmeny byly poté kultivovány v bioreaktorech v médiu obsahujícím kombinaci odpadních substrátů. Kultivace mikrořas rodu Desmodesmus, Scenedesmus, Chlorella, Coccomyxa, Chlamydomonas, Botryococcus se zabývaly optimalizací jednotlivých komponent média a aplikací různých stresů s cílem navýšení produkce studovaných metabolitů. V rámci experimentů s extrémofilní mikrořasou Coccomyxa byly provedeny pilotní velkoobjemové kultivace v otevřených nádržích. V závěrečné části byl provedeny pilotní screeningové a velkoobjemové bioreaktorové experimenty zaměřené na možnosti kokultivace karotenogenních kvasinek a mikrořas. Testované kmeny kvasinek byly s rozdílnou úspěšností schopny utilizovat média obsahující hydrolyzované odpadní substráty. Nejlepším kmenem byl Sporidiobolus pararoseus, který na médiích dosahoval nejvyšších produkcí biomasy i sledovaných metabolitů. Z testovaných odpadních substrátů byla nejlepší kombinace odpadního fritovacího oleje a hydrolyzátu kávové sedliny. Úspěšná optimalizace složení hlavních komponent minerálního média vedla k zvýšené produkci studovaných metabolitů. Největší vliv měl optimální poměr P/N a aplikace oxidačního stresu. Nejlepších výsledků dosáhly mikrořasy rodu Desmodesmus a Scenedesmus. Velkoobjemové kultivace Coccomyxy onubensis potvrdily rezistenci kultury proti kontaminaci vnějšími vlivy a schopnost růstu za vysoké teploty a intenzity světelného záření. Kokultivační experimenty potvrdily schopnost symbiotického růstu kvasinek a mikrořas. Nejlepších výsledků dosahovaly všechny testované kvasinky s mikrořasami rodu Demsodesmus a Scenedesmus a v menší míře i rodu Coccomyxa.
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Optimization of cultivation of carotenogenic yeasts on mixed waste substrates
Holub, Jiří ; Mikulíková, Renata (referee) ; Márová, Ivana (advisor)
The master thesis addresses the issue of cultivation of selected strains of carotenogenic yeasts on waste materials of the food industry using a laboratory bioreactor. Carotenogenic yeasts are able to produce highly valuable metabolites during cultivation, which are located predominantly in the lipid part of the cells. Particularly, they are carotenoids, ergosterol, coenzyme Q and fatty acids. The thesis is divided into two main parts, the theoretical part and the practical part. The theoretical part describes individual yeast strains, types of waste materials, produced metabolites and methods of their analysis. The experimental part deals with the processing of waste materials of the food industry, specifically animal fat, whey and spent coffee grounds into the form of substrates usable as nutrition sources for yeast cultivation. Furthermore, cultivations focused on the recovery of the monitored metabolites and their analysis by using HPLC/PDA and GC/FID assemblies were studied as well. The yeast strains Rhodotorula mucilaginosa (CCY 19-4-6), Rhodotorula kratochvilae (CCY 20-2-26), Rhodosporidium toruloides (CCY 062-002-001), Sporidiobolus pararoseus (CCY 19-9-6) a Cystofilobasidium macerans (CCY 10-1-2) were used in this work. As one of the best producing strains Sporidiobolus pararoseus (CCY 19-9-6) was found, which achieved very high productions of carotenoids, coenzyme Q and ergosterol.
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Metabolic adaptation of selected microalgal strains on various nitrogen sources
Kodajek, Matěj ; Šimanský, Samuel (referee) ; Szotkowski, Martin (advisor)
Microalgae and cyanobacteria are photosynthetic organisms that, together with other microorganisms (yeast, bacteria), are used in industry, because they produce a wide range of interesting organic substances. This thesis deals with the metabolic adaptation of microalgae Scenedesmus obliquus, Scenedesmus acutus, Scenedesmus dimorphus, Chlamydomonas reinhardtii, Coccomyxa sp. and cyanobacteria Synechococcus nidulans, Arthrospira maxima, Limnospira maxima, Anabaena torulosa, which were cultivated on various nitrogen sources. These strains are descibed in the theoretical part including the metabolites and their applications in industry. The experimental part describes all chemicals, aids, devices and methods used for cutivation and analysis of microorganisms. The aim of the study was to compare and find out which nitrogen source in the BBM and SPIRULINA medium is the most suitable for a particular strain in terms of production of total biomass and composition of individual metabolites. Sodium nitrate, ammonium sulfate, glycine, urea and whey protein were used as nitrogen sources. The content and composition of lipids in the biomass was determined by gas chromatography. The content of ubiquinone, carotenoids and chlorophylls was determined by liquid chromatography.
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Metabolic adaptation of carotenogenic yeasts to nutritional stress induced by selected wastes of the food industry
Plhalová, Žaneta ; Šimanský, Samuel (referee) ; Szotkowski, Martin (advisor)
This bachelor thesis deals with the production of carotenoids and lipids by selected strains of carotenogenic yeasts cultivated on waste substrates of the food industry. The production properties of the yeast strains Sporidiobolus pararoseus, Sporidiobolus metaroseus, Cystofilobasidium macerans, Rhodotorula mucilaginosa, Rhodotorula kratochvilovae and Rhodosporidium toruloides were studied. The cultures were cultivated by using waste products of food industry: waste frying oil, coffee oil which was obtained by extraction from coffee grounds and waste animal fat from rendering plants. The work consists of two main parts. The theoretical part describes the individual genera of yeasts, monitored metabolites, waste products and methods of analysis of monitored metabolites. The experimental part is focused on the processing of used waste substrates, cultivation of yeasts, acquisition and processing of biomass and analysis of metabolites. Rhodosporidium toruloides and Sporidiobolus pararoseus strains were the most suitable for the production of carotenoids on waste substrates. The highest production of lipid substances was measured in the strain Sporidiobolus pararoseus.
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Application of physical and chemical stress factors in different stages of growth to autotrophic microorganisms
Sniegoňová, Pavlína ; Byrtusová, Dana (referee) ; Szotkowski, Martin (advisor)
Nowadays, the demand for metabolites produced by microalgae and cyanobacteria is still growing, due to their positive effects on the human body and health. These metabolites include mainly carotenoid pigments, that have antioxidant properties, which very attractive to consumers. These substances are mainly used in food supplements; however they are also used in the pharmaceutical industry. These substances include, for example, -carotene, but also chlorophylls, which are significantly involved in the photosynthesis process. Other important metabolites are lipids, which are part of cellular structures and serve a number of other functions. The theoretical part focuses on the description of algae and cyanobacteria and their cultivation. Furthermore, the influence of stress factors on the production of biomass and metabolites. Metabolites are then characterized in terms of properties and biochemical pathways leading to their production. Subsequently, there are mentioned evaluation techniques used to characterize the cultivated biomass. In the result part, the production properties are compared depending on the selected stress factors.
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