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Use of selected fluorescence techniques to characterization of microbial cells
Vaněk, Martin ; Pekař, Miloslav (referee) ; Márová, Ivana (advisor)
Carotenoids are one of the most abundant natural pigment. Furthermore, they have great potential for wide industrial utilization. Nowadays, prefered (and cheaper) synthetic method counters progressively improved biotechnological production. Far more cheaper production should be reached via implementing methods of on-line optimization, such as flow cytometry. In this thesis the dependence of yeasts autotofluorescence intensity on carotenoids concentration in dry mass was studied. The dependence was established by means of linear regression. Moreover, the dependence between light scatter and the amount of dry mass was established by means of model utilizing forward scatter, side scatter and number of cells in unit volume. Yeasts' cells was also studied under the lenses of laser scanning confocal microscope. The goal was to localize carotenoids inside cells. Experiments undertaken suggest that they are localized inside the lipidic granules, but more experiments are required for a proof.
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Preparation and testing of application forms for natural antimicrobial substances
Vajglová, Klára ; Bokrová, Jitka (referee) ; Márová, Ivana (advisor)
The goal of this bachelor thesis was the study of antimicrobial and antioxidant effects of some herbal extracts: cloves, nettle, sage, chamomile, rosemary, elderberry, lavender and also lysozyme. Further, herbal extracts effect on Gram positive and Gram negative bacteria were compared. In the theoretical part were briefly characterized lysozyme, polyphenols, flavonoids and selected strains of microorganisms. In the experimental section characterization of herbs with antimicrobial and antioxidant effect was proved. Particles containing herbal extracts were prepared and encapsulation efficiency was determined. In some of tested herbs encapsulation efficiency of up to 80% was reached. Stability of the prepared liposomes was tested in the model digestive conditions and after 1 week and 1 month incubation period in model foods. Antimicrobial activity was tested on selected representatives of Gram negative and Gram positive bacteria. Gram positive bacteria were more sensitive to herbal extracts and liposomes containing encapsulated herbs, while lysozyme exhibited higher effect to Gram negative bacteria. Prepared particles with encapsulated antimicrobial extracts are especially suitable for use in the food industry.
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Preparation and use of selected nanoparticles in cosmetics
Hoová, Julie ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The Bachelor thesis is focused on preparation and use of selected nanoparticles in cosmetics. The theoretical part deals with general characterization and use of caffeine, phenolics and vitamin C in cosmetics. Further, characterization of liposomes and possibilities of encapsulation of active substances gained from natural sources were introduced. In experimental part liposomes with encapsulated active substances were prepared by three different techniques. Encapsulation efficiency was determined by HPLC/PDA, phenolics spectrophotometrically and vitamin C by titration. Prepared particles were monitored for size and stability by dynamic light scattering. Furthermore, long-term stability of liposomes in model conditions was evaluated as the amount of the active substances released during incubation. In this work was found out that from all tested samples co-encapsulated liposomes are probably the best application form which can be used in face lotion or gel. Co-encapsulated liposomes exhibited relatively high encapsulation efficiency, good stability and thanks to quality of active substances they meet customer’s requirements of cosmetic industry.
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Production of selected metabolites by carotenogenic yeasts cultivated on food wastes
Turková, Lucie ; Hlaváček, Viliam (referee) ; Márová, Ivana (advisor)
The production of selected metabolites, especially carotenoids, ergosterol and coenzyme Q was observed in four red yeast strains (Cystofilobasidium capitatum, Rhodotorula glutinis, Sporobolomyces shibatanus a Sporobolomyces roseus) cultivated on viticulture waste substrates such as skins and stem. In these substrates the content of carbohydrates, fat, tannins and polyphenols was analysed. Cultivations were done also on chemically and enzymatically hydrolysed substrates. For enzyme hydrolysis the extracellular enzyme cocktails of the fungi Phanerochaete chrysosporium and Penicillium lilacinum were used. The highest production of metabolites was observed in S. roseus cultivated on stem hydrolysed by enzymes from P. chrysosporium. The yield of -carotene reached 616,1 g/g dry mass, carotenoids 753,6 g/g dry mass, ergosterol 415,5 g/g dry mass and coenzym Q 77,9 g/g dry mass. S. roseus reached very high yields also in the other series of cultivation. The lowest production of metabolites was observed in S. shibatanus. The yield coefficients of biomass and -carotene were calculated based on consumption of reducing sugars.
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Characterization of cereal products with fruit component for baby nutrition
Pražáková, Jana ; Lichnová, Andrea (referee) ; Márová, Ivana (advisor)
Present bachelor thesis deals with the characterization of milk cereal products intended for baby food. The theoretical part focuses on the morphology of cereal grains, fortification, an overwiev of supplementary components used to fortification and their properties. For the practical part eight kinds of cereal dairy products of three different producers were chosen. In these products total polyphenols, total flavonoids, reducing and total saccharides and proteins were analyzed. The highest level of polyphenols was measured in product Sunar multicereal. The highest amount of flavonoids (more than 50 % of total phenolics) was found in Nestle fruit product. The highest level of total and reducing saccharides contained Nestlé honey flavour. High amount of proteins was found in products Sunar with multicereal flavour and pears with biscuits. Using HPLC/UV-VIS analysis vitamins C and E were determined. The highest content of vitamin C contained Nestlé with cookie lavor, while high vitamin E level was found in Sunar pear flavor and biscuits. Using HPLC/RI individual saccharides were examined. In all products high amount of lactose was found, mainly in the products of Sunar. Nestlé products contained mostly glukose, while in products Hami sucrose was detected as the most abundant saccharide.
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Cultivation of carotenogenic yeasts on waste substrates usinf solid state fermentation
Rapta, Marek ; Petrik, Siniša (referee) ; Márová, Ivana (advisor)
Semi-solid fermentation is an eco - friendly technique more and more used for production of significant metabolites or enriched biomass at low entrance cost and low comsumption of water. The problem of this technique might be the right choice of microorganism able to grow and produce at low water activity. This work is a pilot study of three red yeast strains – Rhodotorula glutinis, Cystofilobasidium capitatum and Sporobomomyces roseus cultivated by semi-solid state fermentation (semi-SSF). Yeasts were cultivated in series of production media with gradual reduction of cultivation water content. Pasta and hydrolyzed pasta were used as source of sacharides, glucose served as the carbon source in control media. All studied strains we able to grow and produce observed lipidic metabolites also at low water contents. Production of carotenoids and sterols was higher in semi-solid media. Cystofilobasidium capitatum strain was identified as the best producer of carotenoids and biomass. This strain was cultivated on hydrolyzed pasta media with 40% water content. Semi-SSF seems to be an adequate technique for selection of strains having over-productive properties. Observed metabolites production in yeast cells can be optimized by choosing the appropriate production media and water activity.
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Monitoring of cereal products uptake in a sample of Czech population
Vlachová, Martina ; Hurtová, Jana (referee) ; Márová, Ivana (advisor)
This bachelor’s thesis was focused on monitoring of consumption of cereal products and analysis of some of their active components. In theoretical part an overwiev of cereals and their composition was introduced. In experimental part a set of analytical parameters was obtained in selected cereal products: total phenolics, total flavonoids and antioxidant activity. The highest level of these active substances was detected in products containing chocolate and forest fruit. Further, 161 completed consumer questionnaires were collected. From these data common knowledge in consumers, their shopping conventions and individual products preferences were evaluated.
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Influence of polysaccharides on caffein absorption from beverages
Osecká, Karolína ; Márová, Ivana (referee) ; Pekař, Miloslav (advisor)
This bachelor thesis was focused on investigation of the interaction between the polysaccharide sodium hyaluronan (HA) and the alkaloid caffeine (CAF). The reason why this thesis was focused on study of aqueous solutions of hyaluronan with caffeine is that presence of polysaccharide can slow down the realising from hyalurona-caffeine complex. At first, it was determined whether the photophysical properties of caffeine reacts in different polarity solvents. Based on these results was studied the interaction of caffeine and hyaluronan using the fluorescence. Emission and excitation spectra of caffeine were observed. The HA of molecular weight of 1.7 MDa were chosen for the study of the interactions with the caffeine of concentration the caffeine contained in coffee. The results of interaction that would result to influence the emission properties of CAF, not show. In acidic solutions leading to the formation of various shapes of HA and by centrifugation it was determined whether HA in this form interacts with CAF, which was shown to decrease the absorbance of the supernatant in the CAF. The solution and the mixtures were measured in four types of environment in hydrochloric acid solutions at different pH. The influence of constant addition of caffeine and the HA of molecular weights of 1.7 MDa with different concentrations in different acidic solutions. As a part of the work was to determine how hyaluronan behaves in strongly acidic solution. And then determine the pH in model gastric juices after intake of Coca Cola and Kofola. The obtained results can be used as for description of the behavior of caffeine in the presence of selected polysaccharide.
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Antimicrobial peptides and their application into foods
Janhuba, Filip ; Petrik, Siniša (referee) ; Márová, Ivana (advisor)
The presented thesis is focused on a study of antimicrobial peptides and their potential applications into foods. Two types of antimicrobial peptides were studied in detail - bacteriocin nisin and antimicrobial enzyme lysozyme. In theoretical part an overview of antimicrobial peptides used in food industry was introduced. The experimental part of this work was focused on optimization of methods for the determination of antimicrobial peptides concentration. Commonly used spectrophotometric methods for quantitative analysis of total proteins as well as high performance liquid chromatography were tested. To determine the molecular weight and purity a modified SDS-PAGE with the addition of urea and tricine buffer was performed. Further, encapsulation of nisin into liposome was tested. Encapsulated particles were prepared by three different procedures: by ultrasonication, ethanol injection and TLE. Particles stability, distribution and size were studied by dynamic light scattering. Stability of encapsulated peptides in model conditions and model foods was studied too. Finally, antibacterial properties of the selected peptides using test system Bacillus subtilis were studied. Nisin already showed antimicrobial effect at low concentrations - about 1 µg/ml. The inhibitory effect of nisin was more evident in liquid cultures.
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Aggregation processes in amphiphil-polymer system
Šejnohová, Michaela ; Márová, Ivana (referee) ; Pekař, Miloslav (advisor)
This Bachelor´s thesis is focused on the exploration of polyelectrolyte-surfactant system by fluorescence spectroscopy with pyrene as fluorescent probe. Cationic cetyltrimethylammonium bromide (CTAB) was used as a surfactant. Native hyaluronan was used as polyelectrolyte. Hyaluronan was used in one constant molecular weight and one constant concentration. The whole system was prepared in an aqueous environment. Interactions were studied in area between critical aggregation concentration (CAC) and critical micelle concentration (CMC). The influences of the concentration of pyrene and the effect of the way of the samples preparation on aggregation behavior were also studied. It was found that above the critical aggregation concentration exists an area where stable properties of aggregates can be found. The concentration of pyrene has a major effect on the fluorescence properties of the system. Preparation of samples affects the aggregation of the system. At higher surfactant concentrations, above the stable area, was observed phase separation and gel formation.
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