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Comparison of fatty acids profile of selected type of cheese
Pop, Zdeněk ; Diviš, Pavel (referee) ; Vítová, Eva (advisor)
This master thesis deals with the determination of bound and free fatty acids in selected types of cheeses with/without „Protected designation of the Europen Union“ and their mutual comparison. The theoretical part focuses on a milk fat, a brief overview of cheeses, protected designation of the Europen Union, lipids, fatty acids and a possibility of their determination. In the experimental part, free and bound fatty acids profile in 12 selected samples of white mold cheeses was determined. The declared amount of total fat was verified. The cheeses were compared by a content of bound and free fatty acids, the profile of fatty acids was compared based on a type of milk used. Finally, the results were statistically treated by PCA method. The method according to ČSN EN ISO 1735 was chosen for extraction of lipids from the cheese samples. A methanolic solution of boron trifluoride was used for fatty acids esterification. The fatty acids methyl esters were determined by gas chromatography with flame ionization detection.
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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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Analysis of glucans in plant and microbial samples
Vít, Radek ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The aim of the diploma thesis is study of glucans production in selected species of yeasts, algae and plants. Cultivation conditions for yeast strains were performed to gain increased production of glucans under different temperature conditions and in media of different composition. Into the set of tested yeasts species strains Saccharomyces cerevisiae (CCY 6646), Sporidiobolus pararoseus (CCY 19-9-6), Phaffia rhodozyma (CCY 77-1), Rhodotorula glutinis (CCY 20-78-26) and Cystofilobasidium infirmominiatum (CCY 17-18-4) were enrolled. Saccharomyces cerevisiae was cultivated as a control strain because of its verified production of -glucans. -glucans were determinated by the enzyme kit K-YBGL Megazyme. For comparison, algal strain Euglena gracilis (CCALA 349), some species of mushrooms (shiitake, oyster mushroom, garden champignon and Jew’s ear) and cereals (wheat, rye, oats, rice and barley) were analysed too. Further, fatty acid content in the yeast cells was determined by the GC/FID. The best producer of yeast -glucans was R. glutinis CCY 20-7-26, which showed the highest biomass production (12-14 g/l) and also a relatively high amount of -glucans (25-30 %), in cultivation at 15 °C in a medium containing yeast extract in combination with ammonium sulphate. The presence of -glucans has been demonstrated in the microalgae, Euglena gracilis CCALA 349, as well as in samples of higher fungi and cereals.
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Production of beta-glucans by some yeasts and algae
Veselá, Markéta ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
Several yeast strains and microalgae were selected for this diploma thesis. -glucans, lipids, carotenoids, ergosterol and coenzyme Q were determined in selected producers, and the cultivation conditions for yeast strains were optimized to gain enhanced production of -glucans. Microalgae cultivations were carried out according to the instructions of the Collection of Autotrophic organisms (CCALA). Selected microalge strains include Desmodesmus acutus, Dunaliella salina, Arthrospira maxima and Cyanothece sp. Selected yeast species include Rhodotorula glutinis, Cystofilobasidium macerans and Sporidiobolus metaroseus. Edible yeast Saccharomyces cerevisiae was cultivated to compare with other yeast strains because of it's verified production of -glucans. -glucans were then determined by the enzymatic kit K-YBGL Megazyme, carotenoids, ergosterol and coenzyme Q were analyzed by HPLC/PDA and fatty acids were analyzed by GC/FID. The best producer of yeast -glucans was R. glutinis and S. metaroseus, and the best conditions for the production of -glucans and other metabolites was the C/N ratio of 70. Within the microalgae species, only -glucan production was observed, the best producer was D. acutus.
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Determination of lipids and fatty acid profile in a barley caryopsis
Cvrková, Jana ; Vítová, Eva (referee) ; Svoboda, Zdeněk (advisor)
The diploma thesis deals with the determination of lipids and fatty acid profile in a barley caryopsis.(Hordeum vulgare). The theoretical part describes the synthesis of fatty acids and their degradation in plant material, secondly, it described the possibilities of lipid extraction and their determination and the possibilities of determination of fatty acids. In the experimental part method of lipid extraction on automated extractor Fex ®IKA and determination of fatty acids by GC-FID were optimized. For analysis of fatty acids two capillary columns SLB-IL 100 and Supelcowax were compared. Twenty varieties of barley from the year 2008 and twenty varieties from the year 2009 were compared based on the content of lipids and the representation of fatty acids in a barley caryopsis. The diploma thesis was realized in the Research Institute of Brewing and Malting, Plc. in Brno.
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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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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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Production and characterization of immunologically active microbial beta-glucans by yeast.
Gerspitzerová, Nela ; Šimanský, Samuel (referee) ; Szotkowski, Martin (advisor)
The aim of this bachelor thesis is to study the effect of different cultivation conditions on the production of biomass, beta-glucans, lipids, carotenoids and other metabolites in selected strains of carotenogenic yeast. The thesis is divided into two main parts, a theoretical one and an experimental one. Theoretical part contains a characterisation of beta-glucans, their sources and uses, furthermore a description of the selected yeast genera and other metabolites they produce and finally possible analytical methods of these compounds. Experimental part describes the cultivation, analysis and obtained results. During the first experiment, the yeast strains were cultivated in production media with different C/N ratio, during the second experiment, the yeasts were cultivated either at room or at lower temperature with different content of phosphorus in the media. The selected strains of yeast were Rhodotorula glutinis, Rhodotorula mucilaginosa, Rhodosporidium toruloides, Cystofilobasidium macerans, Sporidiobolus metaroseus and Sporidiobolus pararoseus. Increasing C/N ratio generally had a positive effect on the production of lipids, while exhibiting the opposite effect on carotenoids, it did not have an unequivocal effect on the amount of beta-glucans. It was discovered that lowering the cultivation temperature had a significant negative effect on the production of all observed metabolites. Increased content of phosphorus in production media positively influenced the production of biomass and beta-glucans in most strains, on the other hand the highest amount of carotenoids was reached at lower concentration of phosphorus. The amount of phosphorus had almost no effect on the production and the profile of lipids. The highest amount of beta-glucans in biomass was discovered in C. macerans, their biggest production was reached by R. toruloides. The strain S. metaroseus was the most suitable for accumulation of lipids in its biomass. The highest amount of total carotenoids was reached by R. mucilaginosa.
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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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Regulation of production of lipids and lipid compounds in yeasts
Rapta, Marek ; Breierová, Emília (referee) ; Márová, Ivana (advisor)
Oleogenic yeasts under appropriate conditions produce and accumulate lipids and lipid-soluble metabolites in high amounts. This attribute is characteristic also for red yeasts that except lipids accumulate also carotenoids – natural pigments used in food industry and dietary supplements. The aim of this diploma thesis was designed as a comparative screening study of production properties of strains Cystofilobasidium capitatum, Rhodotorula glutinis, Sporobolomyces roseus and Sporobolomyces shibatanus. Choosen carbon sources were glucose and glycerol as waste by-product in biofuel industry. The best production properties were found in Cystofilobasidium capitatum and Rhodotorula glutinis. These two strains produced increased amounts of lipids as well as higher amounts of carotenoids. Strains were tested by FTIR spectroscopy that enables high-throughput, uncomplicated and accurate analysis.
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