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Proposal of systems for targeted transoprt of compounds of sweet taste
Demová, Radoslava ; Hlaváček, Viliam (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on possibility of immobilization sweeteners on polymeric system. In the theoretical part information about sweeteners, carrier systems and the possibility of immobilization technique were reviewed. In the experimental part preparation of nanoparticles of polyhydroxybutyrate (PHB) was optimized. The PHB particles were characterized by dynamic light scattering (DLS). PHB particles were activated by plasma treatment. The surface morphology was studied using infrared and Raman spectroscopy. Immobilization of following substances with a sweet taste was tested: glucose, fructose, sucrose, mannitol, sorbitol and erythritol. From these compounds only sucrose was immobilized on PHB particles successfully. Prepared immobilized particles were exposed to the artificial stomach juice, intestinal juices and bile acids and amount of released sucrose was monitored. Finally, long-term stability prepared particles was measured and also the amount of sucrose released was determined.
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Characterization of some natural substances with antimycotic effect
Plachá, Monika ; Hlaváček, Viliam (referee) ; Márová, Ivana (advisor)
The introduced bachelor thesis is focused on investigation of the effects of natural substances with potential antimycotic effects and characterization of their constituents. As a part of this work, an overview of mycosis, antimycotics and constituents of natural substances were introduced. Moreover, antifungal susceptibility and genus Candida were described. In the end of the theoretical part the liposomes and their characterization were described. In the experimental part general characteristics of some natural substances and antimycotic test were analyzed. Aqueous, ethanol, methanol and DMSO extracts were tested on yeast strain Candida glabrata. Aqueous extract of the cloves, cloves oil and their combination were encapsulated into liposomes. The encapsulation effectivity, long-term stability and their antifungal activity were determined too. Characterization of liposomes was identified by DLS. Finally, application of liposomes to a preparation of pharmaceutical and cosmetic preparations were presented.
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Characterization of antioxidative and antimicrobially active substances in some types of wood
Ďatko, Tomáš ; Hlaváček, Viliam (referee) ; Márová, Ivana (advisor)
The purpose of this bachelor thesis is to analyze antioxidant and antimicrobial active substances in woody plants. The theoretical part deals with characterization of these biologically active substances, the overview of selected types of woody plants and dividing of extraction techniques. Moreover, methods for determination antimicrobial activity were described. The experimental part is focused on preparation of bark extracts of birch, oak and willow using different solvents (water, methanol, ethanol, DMSO) at various parameters (time, temperature). The concentration of polyphenols, flavonoids, tannins and antioxidant activity was set by using spectrophotometrical method. Subsequently antimicrobial activity of extracts was tested against the three microorganisms: Micrococcus luteus, Serratia marcescens and Candida glabrata. The results show that the most antioxidant active substances are found in the willow bark and antimicrobial activity was showed very weakly in selected bark extracts.
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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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Characetrization of selected microbial enzymes
Bradáčová, Kristína ; Hlaváček, Viliam (referee) ; Márová, Ivana (advisor)
This bachelor´s thesis is focused on controlled production and identification of extracellular microbial hydrolytic enzymes by fungi. Theoretical part deals with characterization of selected hydrolytic enzymes, their properties, possibility of production and application. In experimental part the production of enzymes by fungal strains Phanerochaete chrysosporium, Aureobasidium pullulans and Aspergillus oryzae was performed. Cultivation was conducted in submersed mode in mineral medium and in media with waste co-substrates such as wheat bran, sawdust, rapeseed cake (lipids content 2,55 %) and rapeseed cake rich in lipids (9 %). The activity of cellulases, xylanases, amylases, ligninperoxidase, manganese-dependent peroxidase and laccase was monitored during cultivation process and regularly on 3rd, 7th, 10th and 15th day of cultivation. Production of enzymes depended on time and the subsrate type. Cellulases and xylanases were produced mainly on 3rd and 7th day of cultivation, amylases on 3rd and 15th day and lignolytic enzymes on 7th and 15th day. Samples were further separated and analyzed by ultrafiltration, gel filtration and PAGE-SDS electroforesis.
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The negative effect of raw foods due to possible microbial contamination
Šťastná, Martina ; Hlaváček, Viliam (referee) ; Němcová, Andrea (advisor)
Raw diet is the current trend in nutrition, mainly because of the consumption of a balanced diet with a high proportion of health benefits. The basis is the consumption of fresh foods that have not undergone a heat treatment exceeding 42 - 45 ° C. The topic of the bachelor thesis is the general characterization of raw diet, safety of possible microbial contamination and determination of important nutrients and active substances contained in raw products. There are described the most widespread types of meals, approximation of raw diet in terms of its advantages and disadvantages. Also, there are described the most important bacteria, yeasts and molds occurring on fruits and vegetables. By selected methods described in the theoretical part, some nutrients and active ingredients in raw stick samples were determined. The experimental part describes the procedures and principles for the determination of these substances, and last but not least, the real-time PCR methods, agarose gel electrophoresis and the method of sample inoculation for selective solid media that have brought closer the possibility of occurrence of undesirable microorganisms in raw bars and cake. Despite the large number of benefits of raw food consumption and products made from it, it is necessary to take care of its health also in terms of possible contamination by microorganisms. The risk is that the food is not free from unwanted microorganisms during preparation. This is the reason why the raw products are consumed in the shortest possible time after opening, because they become easily a nutrition source for different types of microorganisms due to their composition.
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Determination of Rubisco enzyme content in leaves
Hlaváček, Viliam ; Vránová, Dana (referee) ; Hrstka, Miroslav (advisor)
The aim of the bachelor thesis was to optimize the conditions of the Rubisco enzyme quantity determination by the SDS-PAGE method and the determination of appropriate protein extraction method from plant tissue. Also the concentration range for which the method is linear was determined. Gels with different density ran under different voltages during the electrophoresis were tested for the aims of optimization. Also two different protein extraction methods were used. These were direct extraction to TRIS buffer and extraction with TCA/acetone method. The best results were obtained with protein separation on 8% gel and voltage of 90 V, while the volume of loaded sample was 5 µl. Under these conditions was the Rubisco great subunit band enough separated from the other protein bands, so that the densitometric evaluation provided better quality peaks with good baseline. The linearity of method under these conditions was in concentration range 0,125–1,0 mg of enzyme in 1 ml of sample. Direct extraction of the enzyme with TRIS buffer was less effective than the precipitation of proteins with trichloracetic acid in acetone followed with dissolution of the precipitate. The best buffer for the protein pellet dissolution was the thiourea buffer prepared according to Amalraj et al. [17]..
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Enzymatic hydrolysis of waste cardboard using the SSF method - a source of raw materials for the production of liquid biofuels.
Hlaváček, Viliam ; Stloukal, Radek (referee) ; Gabriel, Petr (advisor)
This master’s thesis discusses the useof enzymatic hydrolysis process of waste cardboard using simultaneous saccharification and fermentation (SSF) as a source of raw materials for production of liquid biofuels. This thesis is based on theses written by Ing. Brummer and Ing.Lepař.Thus, results gained in these works have been used and also further developed. The theoretical part summarizes the reasons for further development of SSF method and discusses, as well, the achievements reached in the processing of lignocellulosic waste materials by the SSF method so far.This section also discusses the general characteristics of lignocellulosic materials and also of the cellulolytic enzymes. It focusses also on individual pretreatment methods of lignocellulosic material and options of increasing the yield of the whole process. The experimental part verifies the particular results reached in previous theses and at the same time a further optimization of the method has been carried out because of the transfer of the whole process into a fermenter. Cardboard was set as the substrate for the experiments as it was evaluated by Ing. Brummer as the best one for enzymatic hydrolysis which was carried out by enzymes from Novozymes®. Parameters such as temperature, pH and kind of used buffer, the loading concentration of substrate and enzymes, were set according to the thesis of Ing. Lepař, which was aimed to their optimization. The SSF process done in fermenter of 2.0 l volume confirmed the previous results and furthermore it has been more effective through optimization of the added inoculum volume. It has been confirmed that the best substrate is cardboard finely grinded by vibrating mill. Also experiments with added nutrients had been done as an effort to increase the ethanol concentration, but these haven’t resulted insatisfying results. The maximal concentration of ethanol was 23,49 g/l, which was achieved after further optimization of various conditions. This result equals to experimental yield of 84,79 %.
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Use of carotenogenic yeasts to production of lipid soluble metabolites
Mariničová, Veronika ; Hlaváček, Viliam (referee) ; Márová, Ivana (advisor)
Carotenoids are fat-soluble chemical compounds that occur as natural pigments in many plants and protect them from sunlight. Lipids are also essential lipophilic substances and they are part of biomembranes. Their main function is primarily to serve as a power supply for the cell, protective function and thermal protection against adverse environmental influences. This bachelor thesis deals with cultivation of selected carotenoid yeast genes, subsequent isolation of carotenoids and other lipid substances, which can be used as a source of potentially beneficial substances for the pharmaceutical or cosmetic industry. The theoretical part deals with the description of carotenoid yeasts, chemical composition and biosynthesis of the metabolites produced, and description of the methods used for their determination. The experimental part is focused mainly on the production of lipid substances by various strains of yeasts using cheap waste substrates and the application of exogenous stress (nutritional stress) to the biotechnological overproduction of selected metabolites using the modification of the production medium. The content of carotenoids, ergosterol and coenzyme Q was analyzed by high performance liquid chromatography with a PDA detector. The lipids accumulated in yeast cells were determined by gas chromatograph with a FID detector. In this work the strains of Sporobolomyces pararoseus, Sporobolomyces metaroseus, Rhodotorula glutinis and Cystofilobasidium infirmominiatum were studied. As waste substrates and carbon sources were used glycerol, which is produced as a waste product in the production of biofuels and whey as an unusable product in dairy technology. The best production on waste substrates was observed in the strains Rhodotorula glutinis and Cystofilobasidium infirmominiatum.
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Production of pigments by yeasts
Gonová, Dominika ; Hlaváček, Viliam (referee) ; Němcová, Andrea (advisor)
Carotenoids are naturally occurring pigments synthesized by yeast, bacteria, filamentous fungi and plants. In recent years, the interest in the study of these pigments and their microbiological production is increasing mainly due to significant biological effects attributed to carotenoids. This work is conceived as comparative study of six carotenogenic yeasts: two strains of species Rhodotorula glutinis, Rhodotorula mucilaginosa, Sporobolomyces roseus, Sporobolomyces metaroseus a Sporobolomyces pararoseus. Their cultivation was carried out in various media where the oxidative stress was applied in the form of hydrogen peroxide and also waste material – egg pasta – was used as a nutrition source. All studied strains were able to use waste substrate as a source of nutrients, in case of the genus Sporobolomyces increasing production of metabolites was observed. Under the oxidative stress, the majority of the studied yeast showed overproduction of carotenoids and accompanying lipid substances. Sporobolomyces roseus CCY 19-6-4 was characterized by the highest production of carotenoids (1923.78 µg/g -carotene) at the egg pasta media, but on the other hand, it showed a significant decline in biomass. As potentially the best producer of biomass (9.85 g/l) and carotenoids (780.24 µg/g of -carotene), the yeast Rhodotorula glutinis CCY 20-2-26 was observed. Pulcherrimin, synthesized mainly by yeast but also by some spore-forming bacteria, also belongs among natural pigments. Recently, its antagonistic effect that was shown against several microorganisms has been intensively studied, as it seems to be very relevant in regard to biological control. The second part of the work deals with the regulation of pulcherrimin production and its antimicrobial activity. As the pigment-producing yeast strains were used Metschnikowia pulcherrima 145, Metschnikowia pulcherrima 147, Mestchnikowia pulcherrima 149, Metschnikowia andauensis 129, which were cultured on various media containing different concentrations of metal ions and hydrogen peroxide as exogenous stress. In the study of pulcherrimin antagonistic effect, gram-positive bacteria Lactobacillus sakei, gram-negative bacteria Escherichia coli, yeast Candida glabrata and filamentous fungi Phanrochaete chrysosporium were used as tested microorganisms. All studied yeasts were able to produce pulcherrimin, and therefore to show antagonist effect, only in media supplemented with iron, while the color intensity was proportional to the concentration of iron. Antimicrobial activity of pulcherrimin was not observed only against bacteria Escherichia coli.
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