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Testing and optimization of protocols for removal of contaminants during DNA isolation from hibiscus blossoms
Brabcová, Martina ; Kubalová, Michaela (referee) ; Fialová, Lenka (advisor)
The Roselle (Hibiscus sabdariffa) is a widely used plant in food industry, especially in tea production. The hibiscus flower is rich in polyphenolic substances, which are a major problem in DNA isolation. This work deals with the modification of the CTAB isolation protocol to ensure concentration and purity of DNA sufficient for its amplification. DNA isolated according to the CTAB protocol did not have sufficient concentration and purity for amplification due to high contamination with polyphenolic substances. A procedure with double incubation of plant tissue in CTAB buffer containing 3% polyvinylpyrrolidone (PVP) was proposed to remove it. These isolates were successfully amplified. The effect of PVP concentration (1-5%) in CTAB buffer on the concentration and purity of isolated DNA was also investigated. PCR results showed that double incubation with CTAB buffer had a greater effect on the removal of polyphenolic compounds than changing the PVP concentration. Subsequently, the modified isolation protocol was also successfully used to isolate amplifiable DNA from commercial tea blends.
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Development of cereal products for specifc nutrition with addition of fruits and vegetables
Růžičková, Magdalena ; Kubalová, Michaela (referee) ; Bendová, Agáta (advisor)
This bachelor thesis focuses on developing gluten-free cereal products for special nutrition. The main focus is on increasing the concentration of phenolic compounds and antioxidants in cereals by adding fruit and vegetable components. The theoretical part includes findings from professional literature on cereals, gluten-free cereals and their chemical composition. A chapter devoted to celiac disease and gluten-free diet was also included. Furthermore, the theoretical part focuses on possible methods of enrichment and also on selected fruits and vegetables used in the experimental part. In the experimental part, five samples of gluten-free cereals were analysed, namely black, red, brown and white rice and corn. The concentration of flavonoids, polyphenols, antioxidants, carbohydrates, proteins, -carotenes, anthocyanins, gluten, -glucans, fatty acids and lipophilic vitamins was determined in the cereal samples. Selected samples were also tested for antimicrobial activity. The black rice sample showed the best results in the majority of the measurements. Selected samples of eight types of fruit (cranberry, raspberry, strawberry, blueberry, blackcurrant, mango, banana, kiwi) and four types of vegetables (broccoli, carrot, green pea, beetroot) were also analysed. The concentration of flavonoids, polyphenols, antioxidants, carotenes, anthocyanins, ascorbic acid and lipophilic vitamins was determined in these samples. Antimicrobial tests were performed on selected fruit and vegetable samples Among the fruit samples, the best results in the majority of measurements were in blueberry and blackcurrant samples. Among the vegetable samples, the best results were obtained from the beetroot sample. From the best cereal, fruit and vegetable samples were created porridges as a final product. In these samples, the concentration of polyphenols and antioxidants was determined. The highest concentration of both compounds was found in the porridge with black rice and blueberry.
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Authenticity of natural plant component in cosmetics products
Kubalová, Michaela ; Fialová, Lenka (referee) ; Němcová, Andrea (advisor)
The purpose of this thesis was to study the authenticity of selected natural ingredients in cosmetic products. These were specifically cosmetic products that contained citruses, mint or lavender. Commercially available isolation kits were used for DNA isolation. The presence of plant origin DNA was verified by PCR method using primers specific for the ITS2 region of plants. The presence of limonene, a significant allergen contained in said plants, was determined in the samples by PCR method using primers for limonene synthase. At the same time, its presence was verified by HPLC method. In addition, two primers were tested for lavender and monitored for their efficacy, with no significant difference in the usage.
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Optimization of cultivation conditions and production of metabolites by microalgae of the genus Desmodesmus and Scenedesmus in photobioreactors.
Kubalová, Michaela ; Mikulíková, Renata (referee) ; Szotkowski, Martin (advisor)
Nowadays, a lot of attention is being paid to the biotechnology industry. Within this industry, are with the help of microorganisms produced various useful substances, such as chlorophyll and carotenoids, which are important antioxidant substances, but also fatty acids, sterols and ubiquinone. This thesis is focused on the cultivation of selected strains of microalgae of the genus Scenedesmus and Desmodesmus. In the first part of the thesis, the algae strains and studied metabolites are described. In the second, experimental part, different types of alcohol were applied to the selected strains as stress conditions, with the aim of increasing the production of intracellular metabolites. Further, fermentor cultivation was carried out, in which a combination of two different alcohols was applied. Subsequently, cultivation using a flat-panel photobioreactor was also carried out, in which light with different wavelengths was used. Several chromatographic techniques were used for the analysis and the results of the analyzes are presented in the experimental part.
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Authenticity of natural plant component in cosmetics products
Kubalová, Michaela ; Fialová, Lenka (referee) ; Němcová, Andrea (advisor)
The purpose of this thesis was to study the authenticity of selected natural ingredients in cosmetic products. These were specifically cosmetic products that contained citruses, mint or lavender. Commercially available isolation kits were used for DNA isolation. The presence of plant origin DNA was verified by PCR method using primers specific for the ITS2 region of plants. The presence of limonene, a significant allergen contained in said plants, was determined in the samples by PCR method using primers for limonene synthase. At the same time, its presence was verified by HPLC method. In addition, two primers were tested for lavender and monitored for their efficacy, with no significant difference in the usage.
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