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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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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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Analysis and characterization of beta-glucans from some natural sources
Vít, Radek ; Hudečková, Helena (referee) ; Márová, Ivana (advisor)
The aim of the bachelor thesis is to test and characterize commercially available products with declared -glucans content. The theoretical part describes -glucans, their possible sources, properties and utilization. Further, the theoretical part is focused on methods for determination of -glucan content. Saccharomyces cerevisiae yeast and oyster mushroom fungus are available as commercial sources of -glucans. Next individual groups of the active substances and principles of the methods used for their characterization are described too. In experimental part of this work the extracts from five commercially available sources of -glucans were prepared and characterized. Antioxidant activity, total polyphenols, proteins and carbohydrates were determined. For selected products, the content of vitamin C using HPLC and the lipid content using GC-FID were determine as well. An enzymatic method was used to determine the -glucan content. Two samples of the highest antioxidant activity were then subjected to antimicrobial and MTT tests.
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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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Determination of beta-glucans and pentosans in raw materials for production of beer and in beer
Piškulová, Lenka ; Sachambula, Lenka (referee) ; Mikulíková, Renata (advisor)
The content of beta-glucans and arabinoxylans in barley grain, malt and wort was monitored. The varieties of barley were Pionier, Libuše, Manta, Tango, KWS Amadora, Kampa, KWS Irina, Francin, Odyssey, Overture, Vendela, Petrus, Laudis 550, Sunshine, Kangoo, Xanadu, Sebastian, Bojos and Malz. These varieties were grown on three different habitats (Uherský Ostroh, Věrovany and Vysoká u Příbramě). The content of beta-glucans and arabinoxylans was also analysed in five different types of beer. In brewing and malting industry the content of beta-glucans and arabinoxylans is mainly monitored because of their negative influence on decanting of wort and filtration of beer. These substances can contribute to turbidity and precipitation in beer and thus negatively effect the stability of beer. The content of beta-glucans in barley grain, malt, wort and beer was determined by FIA method and the content of arabinoxylans was determined in barley grain, wort and beer by Douglas method. The highest content of beta-glucans in barley grain was found in 14/19 varieties at Uherský Ostroh, where the amount of beta-glucan content was in the range of 3,65-5,60 %. The highest beta-glucan content in malt was found in 17/19 varieties at Vysoká u Příbramě, where the beta-glucan content was between 0,15-0,79 % and the highest content of beta-glucans in wort was found in 16/19 varieties also at Vysoká u Příbramě. There was the beta-content in the range of 22-184 mg/l. The results of content of arabinoxylans in barley grain and wort were more balanced. The highest content of arabinoxylans in barley grain was found in 10/19 varieties at Věrovany. The content of arabinoxylans at this habitat was in the range of 3,04-4,56 %. The highest content of arabinoxylans in wort was determined in 8/19 varieties at Uherský Ostroh. The content of arabinoxylans at this habitat was in the range of 542-1040 mg/l. The highest content of beta-glucans in samples of beer was determined in a sample of light lager (207 mg/l) and the highest content of arabinoxylans was found in a samle of wheat lager (1465 mg/l).
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Analysis of beta-glucans in some fungi
Phan, Huong Tra ; Veselá, Mária (referee) ; Márová, Ivana (advisor)
The main goal of this thesis was to analyse the amount of beta glucans in selected mushroom species. The overall characterization of the composition and content of bioactive compounds in mushrooms was studied too. The theoretical part was focused on description of Fungi, especially the Basidiomycota phyla. The overview of fungal morphology and chemical composition was elaborated. Further, theoretical basis of analytical methods used was introduced. In the experimental part, extracts from selected species of mushrooms (shiitake, oyster mushroom, white champignon, brown champignon and jew’s ear) were prepared. Overall group parameters such as polyphenols, flavonoids, antioxidant activity were determined by spectrophotometry. The amount of beta-glucans in the fungi was measured after enzyme hydrolysis using commercial kit. The content and composition of fatty acids was measured by gas spectrometry. Using emission spectrometry (ICP/OES) the presence of heavy metals was analysed in fungal extracts. Cytotoxicity was tested by MTT test on human cell lines. In the last part the antimicrobial activity against Gram-positive bacterium Micrococcus luteus, Gram-negative Serratia marcescens and yeast Candida labrata was tested. In the thesis, a complex characterization of composition and biological effect of some mushrooms was given.
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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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