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Characterization and stabilization of pancreatin
Wurstová, Agáta ; Němcová, Andrea (referee) ; Obruča, Stanislav (advisor)
This work focuses on a study of enzyme mixture pancreatin, its characterization and subsequent encapsulation into liposomes. As a reference proteins bovine serum albumin and trypsin were used. Characterization of pancreatin consisted of two parts. The first part focuses on optimization of methods for the concentration determination by absorption spectrophotometry using basic methods for identifying proteins (Biuret method, Hartree-Lowry method and Bradford method). Moreover, UV spectrums of the protein were measured. As a method for identification of protein´s molecular weight, SDS-PAGE was used. To identify components of pancreatin, LPLC was employed in two modifications, ion-exchange chromatography and size exclusion chromatography. The second part is dedicated to the characterization of pancreatin as enzyme in terms of pH and temperature optimum for the enzyme activities of protease (pH 9, 8 and 50 °C), amylase (pH 7 and 40 °C) and lipase (pH 7 and 50 °C). The last part of this work aimed at an encapsulation of pancreatin into liposomes and DLS analysis of distribution of particles and their zeta potential. Liposomes did not spontaneously release encapsulated enzyme. To confirm that proteins were successfully entrapped into liposomes, their structure was disrupted by application of phospholipase D. In conclusion, liposomes can be utilized as delivery systems for native enzymes.
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Production of lipid substances by Metschnikowia yeasts grown on some waste substrates
Gonová, Dominika ; Márová, Ivana (referee) ; Němcová, Andrea (advisor)
Oleaginous yeasts posses the ability to accumulate increased amount of lipids under appropriate conditions. These microbial lipids vary in the composition of fatty acids which results in their wide application in the biotechnological industry. This master thesis focuses on the lipid production and fatty acids composition from waste substrates by the yeasts Metschnikowia depending on various cultivation conditions. The influence of temperature, the ratio of carbon and nitrogen in medium, and the concentration of different carbon sources was studied. The cheap and easy available waste substrates as glycerol and animal fat were used for the cultivation. The production characteristics of the yeasts were monitored by various technique including gas chromatography, Raman spectroscopy and fluorescence microscopy FLIM. Moreover, the partial optimalization of the pulse field gel electrophoresis was applied in order to characterize the karyotype of the yeasts Metschnikowia. All the studied strains were able to use the waste substrates and at the same time to produce lipids. The amount of lipids and mainly their compositions vary depending on the yeast strain and on the culture conditions. Nevertheless, the ability of the yeasts to produce significant amount of unsaturated fatty acids by manipulation of culture conditions was proved. The maximum lipid yield was achieved by M. pulcherrima 149 on glycerol medium and by M. andauensis 129 on medium containing waste animal fat.
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Determination of athenticity of plant foods by molecular techniques
Plášková, Anna ; Mikulíková, Renata (referee) ; Němcová, Andrea (advisor)
The aim of presented diploma thesis was to determination of authenticity of fruit baby foods for early infant feeding using molecular methods. In the experimental part, isolation kit was used for isolation of plant DNA from fruits (strawberry, apricot, raspberry, apple) and from six commercial fruit products for children. Isolated DNA was characterized and verified using PCR methods with primers specific for plant rDNA (ITS2). Specific primer pairs were designed to amplify DNA for the detection of one fruit species. Primer specificity was assessed with four fruit species. A mixture of fruit puree from the two fruits was used to determine the sensitivity of the multiplex PCR assay. Six commercial fruit products were evaluated to verify the applicability of the multiplex PCR assay. The methodology of molecular detection of fruit DNA by qPCR and multiplex qPCR (duplex) includes approaches, which enable to detect two fruits (strawberry-raspberry, apricot-apple) in one reaction and thus reduces time and money requirements.
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Design of air conditioning in the family house
Němcová, Aneta ; Jelínek, Ondřej (referee) ; Blasinski, Petr (advisor)
The subject of this dissertation is the design of an air conditioning system for a family house. The design contains the actual state, in which a central unit with variable air volume regulators and air supply adjustment is designed. The first version of optimisation offers a simple central ventilation, and in the second version a decentral ventilation for the object. These options were compared based on their purchasing price and additionally from the interior comfort feel point of view. In the next part a long-term measurement of an energy recovery ventilation was performer, and data was processed to evaluate and compare the efficiency with efficiency promised by producer.
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Gait detection and step counting using smartphone
Kočendová, Kateřina ; Vítek, Martin (referee) ; Němcová, Andrea (advisor)
The focus of this thesis is the detection of walking and running, followed by a step count. Testing is performed on signals of regular daily human activity that involve sections of running, walking or no activity. Those signals are logged using accelerometer in smartphone. The exact type of physical activity performed is distinguished by an average, variation coefficient and wave transformation methods. Basics statistic methods are used to quantify the number of steps taken during either walking or running. Algorithm for activity detection and algorithm for step counting are optimized and tested by a set of signals.
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Analysis of some biologicaly active components in natural plant extracts
Kováčová, Ivana ; Márová, Ivana (referee) ; Němcová, Andrea (advisor)
This thesis is focused on the study of the effectiveness of active ingredients from natural extracts such as milk thistle and turmeric. The thesis describes the use of modern methods of analysis and identification of active substances located in plants of milk thistle and turmeric. In the experimental part of thesis, which was preceded by a literature search focused on the biological effects of active substances in the chosen natural extracts, the extracts were characterized according to group characteristics (total assessment of polyphenols and flavonoids, assessment of antioxidant activity, and assessment of vitamin C volume and tocopherol). The next part is focused on the study of the encapsulation of natural active substances into the organic particles: liposomes. Silymarin and curcumin were encapsulated as active substances. The idea of a plant with a single active ingredient is usually erroneous. Plants usually contain hundreds of components which cooperate on a given effect. Therefore, the majority of herbalists believe that a simple plant is an active ingredient. However, some plants are known to be containing a particular active phytochemical. For instance, plants such as the milk thistle, turmeric, or a ginkgo are great examples of the plant containing a particular active phytochemical. In these cases, the active compound is disproportionately more effective than the plant itself. Encapsulation efficiency of active ingredients of Silymarin and curcumin was determined by HPLC/PDA. The aim of the analysis was to observe the behavior of elements during the incubation in surrounding of gastric juice model, and the stability while maintaining the particles in model food.
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Development of a new type of energy drinks containing natural active components
Vysoká, Marie ; Němcová, Andrea (referee) ; Skoumalová, Petra (advisor)
The aim of this thesis is the development and preparation of a new energy drink. The theoretical part of the thesis focuses on the comparison of the drinks available in market, the presentation of the main active ingredients and the proposal of enriching the new energy drink. It also describes the possibility of using an encapsulation and the method used to determine specific compound in energy drink or encapsulated particles. In the experimental part of the work, at the beginning, the main content of energy drinks were characterized, for instance caffeine, sugar, polyphenol and vitamin C. Subsequently the energy drink based on selected types of loose teas that were enriched with the encapsulated extract of acai, acerola, maca, goji and ginseng was designed. All extracts contained large amounts of polyphenols and antioxidants. In addition, acerola extracts contained very high amounts of vitamin C. Two types of particles, alginate particles and liposomes were prepared for encapsulation. For all particles, encapsulation efficiency, size, stability, and antioxidant activity were determined. Long-term stability was then monitored within three months. The particles were also exposed to artificial gastric, pancreatic and bile juice. All tested particles showed very good encapsulation efficiency. In terms of long-term storage stability in an aqueous environment. Particularly liposome particles, which also showed high stability by zeta potential measurement, were suitable. On the contrary, in case of alginate particles, when stored in an aqueous medium, the release of active substances occurred. Finally, an energy drink was created and subjected to sensory analysis. The best energy drink was based on black or green tea with alginate particles acai, goji and ginseng.
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Use of technical hemp in acne preparations
Žáčková, Kristýna ; Kovalčík, Adriána (referee) ; Němcová, Andrea (advisor)
Hemp belongs among important plants with a long industrial tradition. However, its first use was originally for healing. In medicine, it is mainly applied thanks to the content of many biologically active substances, such as cannabinoids. This thesis is concerned with the use of hemp in acne preparations. The theory focuses mainly on the characterization of hemp and its active substances. The other chapters deals with cosmetic preparations and acne itself, and Propionibacterium acnes as well, which is involved. Within the experimental part, the extracts of two hemp varieties (Finola and Bialobrzeskie) were characterized in terms of antioxidant and cannabinoid presence. The cosmetic preparations with hemp extract content were prepared including a complete cleansing set for acne skin, concurrently commercial products of this sort were purchased. In cosmetic preparations and the extracts their inhibitory effect against Propionibacterium acnes was studied as well. Also Micrococcus luteus, Escherichia coli and Candida glabrata were tested. The results show that dried hemp flower extracts contain the greatest amount of active substances, so they have a higher antioxidant and antimicrobial effect. The presence of individual cannabinoids differed mainly depending on the variety. All hemp extracts had effect against gram-positive bacteria, in cosmetic preparations it was lower. Nevertheless hemp antibacterial effect has been proven.
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Application of microbial oils in cosmetics
Reichertová, Klára ; Uhlířová, Renata (referee) ; Němcová, Andrea (advisor)
This bachelor thesis is focused on the application of microbial oil in a cosmetic product. Eight strains of yeasts of the genus Metschnikowia were examined, namely Metschnikowia pulcherrima 145, Metschnikowia pulcherrima 147, Metschnikowia pulcherrima 149, Metschnikowia andauensis 129, Metschnikowia andauensis 1241, Metschnikowia sinensis 1244, Metschnikowia zizyphicola 1247 and Metschnikowia shanxiensis 1250 for production of microbial oil. Substrates made of technical hemp (hemp flour, hydrolyzate and a mixture of leaves and flowers) were used for cultivation. The cultivation was performed at reduced temperature with the advantage of lipid production. The composition and ratio of fatty acids were determined by gas chromatography According to these resulst was choosen the best lipid producer and the most abundant substrate. This was the strain Metschnikowia zizyphicola 1247 and the substrate of hemp flour. At the end of the practical part was prepared a cream, into which the microbial oil was mixed. Finally, an evaluation of the oil with regard to its use in cosmetics and a comparison with vegetable oil with the same fatty acid profile are given.
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Microbial production of lipid substances using waste substrates
Árendásová, Veronika ; Szotkowski, Martin (referee) ; Němcová, Andrea (advisor)
This bachelor’s thesis examines the issue of lipid production, done through selected Metschnikowia strains gained from waste starch substrates. Produced lipids are extensively used in biotechnological processes. Cultivation utilised waste starch substrates, left over from food production, as they constituted an available and economical material. The study also monitored the growth of individual Metschnikowia strains in laboratory and reduced temperatures, using an optimal medium. The yeast’s production properties were analysed by means of gas chromatography. All selected Metschnikowia strains were able to produce lipid from waste substrates. Lipid production increased when a mixture of glucose and waste substrates was used. The amount of lipids and the share of fatty acids strongly depended on the strain and cultivation conditions. In all examined strains, monounsaturated fatty acids were represented most strongly. The Metschnikowia pulcherima 149 adapted best to waste substrates.
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