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Proteome and Metabolome Changes in Selected Organisms under Stress
Halienová, Andrea ; Kráčmar, Stanislav (referee) ; Čertík, Milan (referee) ; Šucman, Emanuel (referee) ; Márová, Ivana (advisor)
Living conditions of every organism are influenced by various factors at this time. Some of them have positive effect on organism, some negative. Basic condition for surviving is the ability to resist and adapt to changing metabolic and living conditions. Every single stress effect can lead to changes in metabolism but organisms have ability to develope sufficient mechanisms for stress response. Some of them are similar for all living organisms (enzyme production, endogenous primary stress metabolites) some of them are specific for certain organism or stress type. Cell stress response can be observed on different levels (proteomic, genomic, metabolomic). In proper conditions it can be used indrustrially. In this work, influences of various stress factors were studied. These factors were applied on selected organisms – carotenogenic yeast and plant materials. Yeast stress response was induced by osmotic and oxidation stress factors. Changes on proteomic level and in production of selected secondary metabolites were observed. Proteome was analyzed by 1D and 2D electrophoresis with subsequent analysis of proteins by mass spectrometry. Yeast strain Rhodotorula glutinis CCY 20-2-26 showed the best adaptation to stress factors, which was moreover accompanied by overproduction of carotenoids. This finding can be premise for next industrial production of carotenoids. In plant samples predominantly enzymes and metabolites involved in antioxidant response were studied.
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Determination of basic nutritional parameters in edible insects
Korček, Jakub ; Štursa, Václav (referee) ; Diviš, Pavel (advisor)
In this bachelor thesis is dicussed nutritional value of chosen insect species. Analysed samples consisted of worms, crickets and locusts. Analysed parameters were: total nitrogen, crude protein, total lipids and content of chosen elements. Total lipids were, after homogenization, determined by Soxhlet method, using diethyl ether as a solvent. Prior to protein and element determination, samples did undergo Kjehldal digestion process, using sulfuric acid. Weiniger catalyst was only added to samples digested for protein measurement. Element content was determined by optical emission spectrometry with inductively coupled plasma (ICP-OES). Crude protein was determined by Kjeldahl method. The highest protein content was determined in crickets. (75 ± 5) %. Locusts contained the highest lipid content (33 ± 2) %. We also found out, that samples were rich in calcium, potassium, manganese and zinc, which are all important elements for the proper functioning of our body. The results of this thesis prove, that insects can present nutritionally important food.
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Fortification of bakery products with protein isolate from wheat bran
Bidmonová, Karolína ; Vítová, Eva (referee) ; Pořízka, Jaromír (advisor)
Proteins, as the major structural component of all cells and tissues, along with fats and carbohydrates, are a key macronutrient in human nutrition. An insufficient protein intake in the diet can lead to health problems and increasing protein malnutrition. Food fortification plays an important role in eliminating this protein deficiency. Wheat products, as a widely consumed food, seem to be a suitable commodity for protein fortification. Wheat bran was selected as a suitable source of protein due to its availability, protein content and relative simply method of a protein isolation. In the experimental part, the protein was extracted from wheat bran and used to prepare the mixtures. The mixture contained T530 flour and the 5%, 10% and 15% addition of the wheat bran protein isolate. From these mixtures, doughs were prepared and were subjected to chemical and rheological analysis. Finally, wheat baguettes with the addition of wheat bran protein isolate were made. The texture and taste properties of the baked products were verified by sensory analysis. The mixtures of flour and whey protein and a sample of unriched flour were also analysed to compare the results. Applied tests have shown that the addition of protein influences not only nutritional and chemical but also rheological properties. The addition of wheat bran protein isolate had a negative impact on the taste properties of the baguette Higher addition of BPI caused the increase of the bitterness of products. Taste optimization could be a topic for future study.
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Analysis of czech garlic varieties
Fähnrichová, Nikola ; Vespalcová, Milena (referee) ; Diviš, Pavel (advisor)
This master thesis deals with analysis of Czech garlic varieties in comparison with two varieties from abroad. Alliin and carbohydrates were determined by high performance liquid chromatography with diode-array detector or evaporative light scattering detector. Crude proteins were calculated from total content of nitrogen that was determinated by Kjeldahl method. Elementary analysis was performed by inductively coupled plasma optical emission spectrometry. In theoretical part of this thesis is describes botanical characterization and cultivation of garlic, chemical composition of garlic, especially groups of substances which were analysed in this thesis. Principle of used methods is described too. The experimental part of this work deals with sample and calibration solution preparation and process of analysis. Obtained results from analysed substances are summarized in the tables, discussed and compared with results of analysis of other authors. Between Czech and Spanish/Chinese garlic are slight differences. Garlics contain 24,66 g/kg dry weight of alliin. Content of crude proteins in Czech garlic is comparable to Chinese garlic but the Spanish garlic has just half of amount crude proteins. Elementary analysis also includes significant differences. Czech varieties contain higher amount of phosphorus, iron and zinc. On the contrary, content of potassium in foreign garlic is higher than in Czech Republic and the analysed amount is 15 g/kg dry weight.
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Nutritional benefit of cricket flour
Šťastná, Martina ; Zemanová, Jana (referee) ; Němcová, Andrea (advisor)
Edible insects have been part of the diet of many countries across the world for several decades. It is becoming increasingly popular in the europian developed countries, mainly because of the content of nutritionally important substances. The theoretical part of the thesis deals with the characterization of edible insects in terms of nutritional benefits, explains the reasons and method of breeding edible insects. For the analysis of nutritionally active substances, flour of house cricket (Acheta domesticus) was used. A. domesticus belongs to the representatives of insects legalized from 1.1. 2018 on the EU market. Of the significantly nutritional substances, proteins, lipids, fatty acids, fiber and selected minerals were analyzed. In this study, optimalization of vertical electrophoresis SDS-PAGE was performed to determine protein fragment sizes. In the second part the influence of acricket flour addition protein bars was investigated by sensory analysis. In the experimental part that was found that cricket flour contains protein, lipids and fatty acids in significant amounts. It also contains important minerals for human health such as magnesium, potassium, iron or zinc. However, the sensory properties of protein bars for consumers were not very attractive.
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Isolation of pure aminoacids from wheat bran
Sloupová, Klára ; Diviš, Pavel (referee) ; Pořízka, Jaromír (advisor)
Wheat bran is a promising material containing a wide range of useful components, including proteins. In addition, it is produced in significant volumes. Currently, wheat bran is used for the production of energy by combustion and for feed purposes. Gradually, new methods of valorization of this material are being sought. One of the possibilities of using wheat bran is the isolation of proteins, hydrolysis, and separation of selected amino acids. This diploma thesis deals with this issue, it is focused on the recovery of arginine and leucine from a protein isolate. Proteins were extracted from wheat bran by changing the pH. Thanks to the subsequent lyophilization a protein isolate was gained. Prior to hydrolysis of the resulting isolate, a stability test of arginine and leucine amino acid standards was first performed, to which various hydrolysis methods were applied. Acid hydrolysis using a mineralizer, which was applied to the protein isolate, was proved to be the most effective. This was followed by the derivatization of the hydrolysates with OPA and analysis of the resulting hydrolysates by high-performance liquid chromatography with UV-VIS detection. Then, suitable adsorption and desorption conditions were optimized. It was found that the time dependence does not affect the amount of adsorbed material on the sorbent. Therefore, an application time of 15 minutes was chosen. While optimizing the amount of used standard, it was found that the optimal weight was 0.25 g of sorbent. The selected conditions were applied to the protein hydrolyzate. Two fractions were obtained by the separation of selected amino acids due to the change in the pH of the citrate buffer. After the application of this procedure, 0.26 g of arginine and 0.82 g of leucine were obtained from one kilogram after evaporation. From evaporation two, 1.01 g of arginine and 0.25 g of leucine were obtained after evaporation.
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Methods for prediction of secondary structure in proteins
Hoštáková, Nina ; Provazník, Ivo (referee) ; Maděránková, Denisa (advisor)
The examination of protein structure is crucial in determining protein function in organism. This work deals with the issue of 1D, 2D and 3D structures, into which are proteins organized in space. Emphasis is placed on secondary structure, which can be predicted directly from the amino acid sequences and then used for the estimation of spatial structure. On this procedure are focused computational methods, using algorithms that convert the order of amino acids into the order of preferences for secondary structures. To direct determination of the structure by creating structural models is devoted chapter Experimental Methods (NMR spectroscopy, RTG crystallography). The main aim of this work is practical realization of protein secondary structure prediction method. The created program is supplemented by graphical user interface. In the final part the results of the program based on Chou- Fasman method are compared to the outputs of freely available softwares from the Internet.
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Analysis of Protein Structure Visualisation Tools
Klemšová, Jarmila ; Jaša, Petr (referee) ; Burgetová, Ivana (advisor)
In this thesis we analyze several tools for protein structure visualisation. We describe the structure of the proteins, the databases for storing protein structures and their data formats. Next part of the thesis, provides detailed information about selected visualisation tools and some scripts for them. The thesis also include an example of visualisation of the selected protein.
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Detection of Homologous Enzymes
Gajdoš, Pavel ; Burgetová, Ivana (referee) ; Martínek, Tomáš (advisor)
Tato práce se zabývá vyhledáváním homologních enzymů v proteinových databázích, jejímž cílem je navrhnout nástroj poskytující takové vyhledávání. Čtenář se seznámí se základní teorií týkající se proteinů, enzymů, homologie, ale také s existujícími nástroji pro vyhledávání homologních proteinů a enzymů. Dále je popsáno ohodnocení nalezených existujících nástrojů pro vyhledávání homologních enzymů. Pro potřeby vyhodnocení byla vytvořena datová sada spolu s algoritmem pro vyhodnocení vyýsledků jednotlivých nástrojů. Další částí práce je návrh a implementace nové metody pro vyhledávání homologních enzymů společně s jejím vyhodnocením. Jsou popsány dva algoritmy (One-by-One a MSA) pro vyhledávání homologních enzymů, jejichž porovnání ukazuje, že MSA algoritmus je zanedbatelně lepší z hlediska přesnosti než One-by-One algoritmus zatímco z hlediska rychlosti vítězí One-by-One algoritmus.
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Effect of the dry hopping of the stability of beer foam
Benda, David ; Štursa, Václav (referee) ; Pořízka, Jaromír (advisor)
This diploma thesis deals with the influence of dry hopping on the stability of the beer foam. It was observed how this effect affected general characteristics of beer, the content of hop bitter acids, proteins, phenolic compounds, selected micro- and macro elements, organic acids and the stability od the beer foam. Theoretical part characterises raw the materials for beer production, the substances affecting the stability of beer foam and the beer brewing process. Furthermore the beer foam is described froam a physical point of view together with the steps of its formation and drainage as well as other phenomena that lead to its disappearance. A detailed description of used analytical methods is included. The experimental part illustrates the preparation of the model beer and the implementation of dry hopping with the Czech hop variety called Kazbek. Dry hopping was carried out on day 3, day 5 and day 10 with a 3 g/l and a 6 g/l hop dose. The prepared beer was subsequently analysed for general characteristics, the beer foam stability, bitter hop acids, phenolic compounds, protein, selected micro- and macroelements and organic acids. Based on the measured data, there were found correlations between the observed beer parameter and the stablity of the beer foam. The results revealed that dry hopping in beer increases the content of the phenolic compounds and bitter substances, but its effect on the beer foam stability is negative.
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