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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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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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Optimization of extraction of pigments from yeast and algae cells
Šimanský, Samuel ; Mikulíková, Renata (referee) ; Márová, Ivana (advisor)
The presented diploma thesis deals with the extraction and storage stability of lipophilic pigments produced by selected strains of yeasts and algae. In this thesis, there was studied the influence of the selected solvents on the efficiency of extraction, as well as the effect of ambient temperature on the stability of the pigments during storage. The work is divided into two main parts, theoretical and practical part. In the theoretical part knowledge about algae, yeasts and their cultivation parameters is summarized. Furthermore, some information regarding the selected lipid metabolites, their properties and possibilities of application in various branches of industry is mentioned. The practical part deals with the preparation of extracts and stability tests. Extracts were prepared from selected biomass samples in solvents suitable for applications in food industry or cosmetics (ethanol and hexane). Subsequently, the long-term stability tests lasting 4 months and short-term stability tests lasting a total of 28 days were performed on these extracts. The pigments were determined by HPLC and spectrophotometrically, the fatty acid content was determined by GC. For the extraction of pigments from biomass, in the most cases ethanol appears to be the optimal solvent. However, for lipid extraction from biomass, hexane appears to be the optimal solvent for a significant number of samples. In most samples, storage in the freezer showed the most favourable effect on pigment stability, but some samples showed comparable stability even when stored in the refrigerator.
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Determination of residual substances from the preparation of colloidal systems based on biopolymers
Borůvková, Tereza ; Vávrová, Milada (referee) ; Mravec, Filip (advisor)
Volatile organic compounds could be isolated and concentrated from liquid solution using methods, which are based on analysis of equilibrium gas phase, because of their volatility. The aim of this bachelor´s thesis is determination the amount of residual tert-butanol, belonging to volatile organic compounds, in the samples after lyophilisation. Samples were prepared by mixing hydrofobic sonde and hydrofilic hyaluronan. For mixing these components it is necessary to add organic solvent tert-butanol, which ensures stable environment. Thanks to its presence mixing of probe and hyaluronan occurs. Consecutive lyopfilisation of the sample leads to the disruption of a water cover of hyaluronan and nonpolar hydrogen atoms, which are hidden in it, can interact with hydrofobic probe. Bonding the probe is important in oncology, where this fact could be used in targeted distribution of pharmaceuticals. However presence of tert-butanol is unacceptable in the final product, mainly in medical applications. The object this study is the minimization of residual tert-butanol amount in final lyophilised cake. The method of equilibrium gas phase analysis (static Head-Space) by gas chromatography with flame ionization detection (GC/FID) was used for the determination of its content.
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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 pigments and lipid substances by microorganisms on waste substrates of the food industry
Hladká, Dagmar ; Němcová, Andrea (referee) ; Szotkowski, Martin (advisor)
The presented study is focused on production of carotenoids, lipids and other substances by carotenogenic yeasts and autotrophic algae. Studied strains were cultivated in media with different composition, including waste substrates from food industry. Studied strains were cultivated under stress conditions to enhance the production of desired metabolites. The theoretical part deals with the information about yeasts and algae, with the information about monitored metabolites such as lipids, carotenoids, ergosterol, ubichinon or chlorophyll. Furthermore, the theoretical part deals with possible methods of metabolite analysis. The experimental part deals with the description of cultivation of yeats and algae. Also experimental part is focused on the description of individual methods. The result part deals with comparition of production of biomass, metabolits and lipids. The selected strains of yeast were Sporidiobolus pararoseus, Sporidiobolus metaroseus, Sporobolomyces roceus, Phaffia rhodozyma and Dioszegia hungarica. The selected strains of algae were Desmodesmus acutus, Desmodesmus quadricauda, Scenedesmus dimorphus and Chlamydomonas reinhardtii. We were optimized conditions for metabolites and lipids production. Optimal medium, which contained coffee hydrolyzate was with carbon to nitrogen ratio C/N 25. Nitrogen in higher concentration had negative effect on production of lipids. The most suitable strain for effective use of nitrogen in different concentrations was Chlamydomonas reinhardtii. The most suitable strain of yeast for effective use of coffee hydrolyzate was Sporidiobolus metaroseus.
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A comparison of metabolic activity and use of microalgae and pigment-formed yeasts
Szotkowski, Martin ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
Carotenoids are natural pigments occurring in plants and many microorganisms, such as algae, yeast and bacteria. They represent the most common group of antioxidants with significant biological effect. Lipids are an essential component of all living organisms. They are the source and the reservoir of energy for organisms, which can also be used in the petrochemical industry for the production of biofuels. This thesis deals with the production properties of carotenoids and lipids by selected strains of yeasts, algae, and canobacteria, in order to apply the acquired knowledge and find cheap suitable alternatives for microbial biotechnological production of these metabolites using waste substrates. Carotenoids, coenzyme Q, and ergosterol in cells were determined by liquid chromatography. The lipid content and fatty acid profile was analyzed by gas chromatography. Cell morphology and localization of selected metabolites were analyzed by fluorescence lifetime imaging microscopy. In this work yeast genera Sporobolomyces, Cystofilobasidium and Rhodotorula were used. As the representatives of the algae and cyanobacteria strains of Desmodesmus, Cyanothece, Chlamydomonas, Synechoccocus and Chlorella strains were studied. In the overall comparison, the yeast strains were more productive than algae and cyanobacteria. The highest carotenoid production was found in S. pararoseus, C. capitatum and R. mucilaginosa cells, while the highest lipid yield was observed in strains of C. infirmominiatum and S. metaroseus.
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Production of lipids by different yeast strains
Ponížil, David ; Byrtusová, Dana (referee) ; Szotkowski, Martin (advisor)
The bachelor's thesis is focused on the cultivation of selected species of yeast. It is proven that these lipids have many uses and can be applied in the production of biofuels or food industry products. Lipids are forming basic biomolecules that are part of biomembranes in living organisms. The theoretical part describes the structure, chemical composition, and nutrition of these microorganisms. Furthermore, this work contains the characteristic not only of the yeast strains used but also of the lipid metabolites. The experimental part is focused on the description of the preparation of the culture medium designed for yeast growth. The total proportion of lipids and lipid metabolites in the biomass was monitored and compared to individual yeast strains grown on culture media with different compositions. The yeast was cultivated on mediums with carbon and nitrogen from different sources and in different ratios. Waste substrates were also used as carbon and nitrogen sources to compare them to conventional sources. The profile of fatty acids produced by selected yeasts was compared to the fatty acids contained in vegetable oils and butter. The series of experiments showed that the largest proportion of lipids in biomass was observed in the strain Rhodosporidium toruloides grown on a waste substrate of animal fat.
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Comparison of growth and productivity of autotrophic algae and red yeasts
Kouřilová, Xenie ; Rapta, Marek (referee) ; Márová, Ivana (advisor)
The submitted bachelor thesis focuses on the comparison of growth and productivity of red yeasts Rhodotorula aurantiaca and Xanthophyllomyces dendrorhous and autotrophic algae Chlamydomonas reinhardtii and Chlorella vulgaris under different physical conditions. The theoretical part contains the characteristics of selected micro-organisms, stress factors caused by temperature, radiation, combination of aeration and radiation and osmotic stress. Furthermore, knowledge on metabolites and the possibilities of their analysis are mentioned. Physical stresses, namely 2%, 3%, 4% and 5% solution of sodium chloride, intense radiation, absence of lighting, 8°C and 30°C, were applied to selected strains in cultivation. Under these conditions, changes in production of carotenoids, fatty acids, ergosterol, ubiquinone and proteins were being observed. A significant inhibitive effect on all the above mentioned micro-organisms was brought about by the use of 5% solution NaCl. The highest production of metabolites with X. dendrorhous was discovered when 4% solution of NaCl was used. R. aurantiaca demonstrated higher substance utilization at 30°C. For the selected algae strains were the used stresses lethal. Within the cultivation process, the possibilities of co-cultivation of yeasts and algae were also tested, which will be the subject of further studies.
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Characterization of individual phases of cultivation of red yeasts
Valentová, Lucie ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
This bachelor thesis deals with the problematic of metabolic activity and morphology of several biotechnologically significant red yeasts - Rhodotorula glutinis, Sporobolomyces shibatanus and Phaffia rhodozyma during the cultivation time. These yeasts were cultivated on glucose medium, during the cultivation samples were taken for the analysis of produced metabolites. The amount of -carotene, astaxanthin, coenzyme Q and ergosterol were monitored. The amount of biosynthesised lipids and fatty acid profile was also investigated. HPLC/PDA and GC/FID were used for the analysis of these metabolites. Yeasts were cultivated 56 hours (they reached early stationary phase). The highest production of carotenoid pigments showed Phaffia rhodozyma (1856.58 g/g), 42 % of this amount represents -carotene. The highest lipids biosynthesis efficiency was reached by Rhodotorula glutinis. To study the inner structure of yeast cells the Fluorescence Life-time Imaging was used, the Autofluorescence of carotenoids and fluorescence of lipid bodies induced by lipophilic dye Nile red was observed. This method is very useful for imaging cell inner structure (membranes, some kinds of organelles). The changes that the cell undergoes during its evolution could be quantified too. The relationship between the level of carotenoid concentration, membrane rigidity and metabolic shift was found.
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