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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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Vegetable adaptogens - Schizandra chinensis
Pospíchalová, Lucie ; Půčková, Helena (referee) ; Kotlík, Josef (advisor)
This thesis deals with plant adaptogens (bioactive substances) in the plant Schisandra chinensis. The theoretical part is focused on characterization of the Schisandra genus, characterization of the Schisandra chinensis as a plant, its properties, occurrence and growing possibilities in the regional climate zone of the South Moravian Region as well as the use of this plant. Furthermore, adaptogens are described from a chemical point of view and their possible methods of isolation from individual parts of plants. The experimental part deals with extraction of fruits and leaves from plant Schisandra chinensis. Further, in the leaves extract was determined chlorophyll a, chlorophyll b and carotenoids by spectrophotometry, where the quantity of individual substances was confirmed for months April – May.
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Production of selected metabolites by yeasts and algae cultivated under stress conditions
Mariničová, Veronika ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
The presented work was focused on the comparison between the production of selected metabolites by carotenogenic yeasts and microalgae cultivated under conditions of external stress. The main metabolites of interest were carotenoids, further lipophilic substances and lipids. Biotechnological overproduction of these metabolites could serve as a source of potentially beneficial substances not only for the pharmaceutical, cosmetic and food industries, but also for the production of third generation biofuels. Recently, there has been a growing interest in biofuels primarily from microalgae, which have a high potential in biofuel production and seem to be a promising source. The theoretical part deals with the description of individual genera of carotenogenic yeasts, microalgae, cyanobacteria, chemical composition of produced metabolites and brief biosynthesis. In addition, individual methods for analyzing the production of the metabolites of interest were described. The experimental part is focused on the comparison of production of carotenoids, coenzyme Q, ergosterols (phytosterols) and lipids by yeasts, microalgae and cyanobacteria. As a source of external stress, temperature, salt and light stress were chosen. The strains of Rhodotorula glutinis, Rhodotorula mucilaginosa, Sporidiobolus pararoseus and Cystofilobasidium macerans were studied from the yeast strains. Microalgae and cyanobacteria were Scenedesmus obliqus, Scenedesmus dimorphus, Chlorella sorokiniana, Chlorella saccharophila, Botryococcus brauni, Synechococcus nidulans and Arthrospira maxima. The yeast and algal strains were optimized for growth, carotenoid and lipid production. Applied salt stress showed a significant liquidation effect on algal and cyanobacterial strains. The thesis also monitored the biological stress, so-called co-cultivation of microalgae and yeasts. Further experiments will be the subject of future work.
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Production of carotenoi by yeasts of the genus Cystofilobasidium
Vavrysová, Alena ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
Carotenoids are important industrial pigments present practically in all living organisms. The aim of presented work is the study of regulation of carotenoid production in yeasts of the genus Cystofilobasidium in presence of exogenous stress factors. Growth curve of C. capitatum exhibited typical two-stage course with prolonged stationary phase similar to other carotenogenic yeasts. Maximal production of biomass and beta-carotene occurred in 103rd hour. Applied stress factors (2-5% NACl, 2-5 mM H2O2, 0,01-1 mM Se(IV), 0,1-5 mM Cr(III)) exhibited no significant influence on biomass production, which reached on average 8-9 g/l. Positive effect was observed in presence of 5mM Cr where 10 g/L of biomass was produced. Beta-carotene formation was positively influenced by many applied stress factors, the highest yield (695 g/g) was reached in presence of 0,1 mM Se(IV). No simultaneous regulation of ergosterol and carotenes was observed in Cystofilobasidium cells. Production properties of yeast strain C. capitatum CCY 10-1-1 wee compared with those of other carotenogenic yeasts of the genes Rhodotorula and Sporobolomyces. C. capitatum produced similar biomass yield as Rhodotorula sp. in presence of salt. Production of beta-carotene by C. capitatum was slightly higher than in Rhodotorula glutinis, but lower than in Sporobolomyces strains which exhibited substantially lower biomass production. Karyotype of C. capitatum is relatively different when compared with karyotype of other carotenogenic yeasts. Based on summary of our results in seems that yeasts C. capitatum exhibit similar physiological as well as production properties as some Rhodotorula strains. Thus, yeasts of the genus Cystofilobasidium could be potentially used to industrial production of carotenoid pigments as well as yeast biomass rich in carotenoids and some biogenic elements.
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Use of autotrophic algae to production of biomass and some metabolites
Chrástová, Nikola ; Kostovová, Iveta (referee) ; Márová, Ivana (advisor)
Chlorophyll and carotenoids are natural pigments and antioxidants. These pigments are suitable for use in the pharmaceutical, cosmetics or food industries. Lipids are another major metabolite of algae and are a potential feedstock for biofuel production. This bachelor thesis deals with the cultivation of selected genera of autotrophic algae, subsequent isolation of these selected metabolites and their application. The theoretical part focuses on the description of the algae, the metabolites produced, the cultivation and the chosen methods. The experimental part deals with the optimization of the cultivation of algae of the genera Chlorella, Chlamydomonas and Desmodesmus, depending on the chosen medium and intensity of illumination, determination and quantification of algal biomass. Further, extraction of lipidic and carotenoid substances and determination by HPLC and GC was performed. In the overall comparison, the best producer of biomass is the algal genus Desmodesmus and the highest production of pigments and lipid substances was reached in algae of the genus Chlamydomonas.
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Simultaneous co-cultivation of selected strains of carotenogenic yeasts and autotrophic bacteria
Blažková, Jana ; Němcová, Andrea (referee) ; Szotkowski, Martin (advisor)
The submitted diploma thesis was focused on the study of co-cultivation of selected microorganisms, which were carotenogenic yeasts and cyanobacteria. The production of selected metabolites was compared in these co-cultivations. The main metabolites monitored were carotenoids, sterols, coenzyme Q10, chlorophylls and lipids. Furthermore, this work focused on the study and possibilities of optimizing the production of lipids and lipid substances in selected strains of carotenogenic yeasts and cyanobacterial species. The theoretical part is focused on the description of yeasts, especially carotenogenic yeasts, cyanobacteria and the chemical composition of the produced metabolites. Microorganisms such as yeast and cyanobacteria contain carotenoids, which are natural pigments and are classified as antioxidants. As antioxidants, they have significant biological effects, such as effects on human health. Coenzyme Q has a positive effect on the functioning of organs in the human body. Chlorophyll is widely used in the food industry as a green dye. Lipids produced by microorganisms contain a large amount of unsaturated fatty acids, which is currently used in cosmetics or pharmacy. The theoretical part also described the individual analytical methods by which the production of the monitored metabolites could be determined. The experimental part is focused on the production of carotenoids, sterols, coenzyme Q10 and chlorophyll, which were determined by HPLC, lipids and fatty acid profile were determined by GC. The determined metabolites are monitored in different types of co-culture partners (carotenogenic yeast and cyanobacteria) in media with different additions of macroelements (P, N and Mg). This was followed by a co-cultivation experiment using waste oils (frying and coffee oil) and a study of the effect of waste oils added to co-cultivations. Co-cultivation experiments confirmed the ability of carotenogenic yeasts and cyanobacteria to grow together. The best results were obtained with Rhodosporidium toruloides and Anabena torulosa, Rhodosporidium toruloides and Arthrospira maxima.
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Potential use of waste lipid substrates and glycerol to production of carotenoids by yeasts
Kostovová, Iveta ; Obruča, Stanislav (referee) ; Márová, Ivana (advisor)
The aim of this study was use of lipid waste substrates and waste glycerol as carbon sources for red yeasts. Six yeast strains, namely Rhodotorula glutinis, Rhodotorula rubra, Rhodotorula aurantiaca, Cystofilobasidium capitatum, Sporobolomyces roseus and Sporobolomyces shibatanus were used for cultivation. All studied yeast strains were able to utilize glycerol and rapeseed oil as a carbon source. Cultivation in oil medium was in most of strains accompanied by decreased production of carotenoids. Increased production of ergosterol, lycopene and biomass was observed in some strains only. Most of yeast strains cultivated in glycerol medium produced increased amount of biomass as well as carotenoids. The best producer of bi-omass and pigments was yeast strain Sporobolomyces roseus CCY 19-6-4 grown in medium containing glucose and glycerol in a 1:1 ratio.
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