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Novel approaches for PHA isolation from bacterial biomass
Dlouhá, Karolína ; Slaninová, Eva (referee) ; Obruča, Stanislav (advisor)
The aim of this diploma thesis was study of the isolation of polyhydroxyalkanoates using a commercial surfactant and selected biosurfactants, which were sophorolipids, coconut soap, lecithin, lauryl glucoside, coco glucoside and cocamidopropyl betaine. PHA was isolated from Schlegelella thermodepolymerans DSM 15344, where the amount of NaCl in the production medium was first optimized. The molecular weight of the isolated polymer was analyzed by SEC-MALS. The largest structural changes were recorded for coconut soap. Possible impurities in the isolated polymer were analyzed by infrared spectrometry (ATR-FTIR). The least protein contamination of the polymer was recorded with coconut soap. From the above biosurfactants, coconut soap, lauryl glucoside and coco glucoside were selected, because the highest purity of PHA was obtained. However, coconut soap had the greatest potential. The isolation temperature and surfactant concentration were optimized for selected biosurfactants. The best results were obtained at 90 °C and a concentration of 5 g/L. Furthermore, the versatility of the isolation method was tested using selected biosurfactants on various thermophilic microorganisms, which were Chelatococcus composti DSM 101465, Schlegelella thermodepolymerans DSM 15264, Tepidimonas taiwanensis LMG 22826 and Aneurinibacillus thermoaerophilus H1.
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Characterization of PHA producing microbial cells by advanced microscopic and cytometric techniques
Dlouhá, Karolína ; Nováčková, Ivana (referee) ; Obruča, Stanislav (advisor)
The aim of this bachelor thesis is to document production of polyhydroxyalkanoates in selected bacterial strains, which were Pseudomonas thermotolerans, Chelatococcus daeguensis, Tepidiphilus thermophilus and Chelatococcus thermostellatus. In the case of the microorganisms Tepidiphilus thermophilus and Pseudomonas thermotolerans, the production of PHA´s has not yet been described, and in the Chelatococcus bacteria, which were analysed in this work, the production has not yet been documented by electron microscopy. In this work, the selected producers were analysed by flow cytometry first, using BODIPY and Nile Red as fluorescence probes. Selected producers, for which was production confirmed, were analysed by other methods, such as fluorescence microscopy, cryo scanning electron microscopy and transmission electron microscopy. In the bacterial culture of Pseudomonas thermotolerans, PHA´s production wasn´t confirmed by the first analysis by flow cytometer. For other microorganisms was production confirmed. Chelatococcus bacteria clearly proved to be better producers. Bacterial cells of Tepidiphilus thermophilus produced smaller granules and in lower amount.
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Novel approaches for PHA isolation from bacterial biomass
Dlouhá, Karolína ; Slaninová, Eva (referee) ; Obruča, Stanislav (advisor)
The aim of this diploma thesis was study of the isolation of polyhydroxyalkanoates using a commercial surfactant and selected biosurfactants, which were sophorolipids, coconut soap, lecithin, lauryl glucoside, coco glucoside and cocamidopropyl betaine. PHA was isolated from Schlegelella thermodepolymerans DSM 15344, where the amount of NaCl in the production medium was first optimized. The molecular weight of the isolated polymer was analyzed by SEC-MALS. The largest structural changes were recorded for coconut soap. Possible impurities in the isolated polymer were analyzed by infrared spectrometry (ATR-FTIR). The least protein contamination of the polymer was recorded with coconut soap. From the above biosurfactants, coconut soap, lauryl glucoside and coco glucoside were selected, because the highest purity of PHA was obtained. However, coconut soap had the greatest potential. The isolation temperature and surfactant concentration were optimized for selected biosurfactants. The best results were obtained at 90 °C and a concentration of 5 g/L. Furthermore, the versatility of the isolation method was tested using selected biosurfactants on various thermophilic microorganisms, which were Chelatococcus composti DSM 101465, Schlegelella thermodepolymerans DSM 15264, Tepidimonas taiwanensis LMG 22826 and Aneurinibacillus thermoaerophilus H1.
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Characterization of PHA producing microbial cells by advanced microscopic and cytometric techniques
Dlouhá, Karolína ; Nováčková, Ivana (referee) ; Obruča, Stanislav (advisor)
The aim of this bachelor thesis is to document production of polyhydroxyalkanoates in selected bacterial strains, which were Pseudomonas thermotolerans, Chelatococcus daeguensis, Tepidiphilus thermophilus and Chelatococcus thermostellatus. In the case of the microorganisms Tepidiphilus thermophilus and Pseudomonas thermotolerans, the production of PHA´s has not yet been described, and in the Chelatococcus bacteria, which were analysed in this work, the production has not yet been documented by electron microscopy. In this work, the selected producers were analysed by flow cytometry first, using BODIPY and Nile Red as fluorescence probes. Selected producers, for which was production confirmed, were analysed by other methods, such as fluorescence microscopy, cryo scanning electron microscopy and transmission electron microscopy. In the bacterial culture of Pseudomonas thermotolerans, PHA´s production wasn´t confirmed by the first analysis by flow cytometer. For other microorganisms was production confirmed. Chelatococcus bacteria clearly proved to be better producers. Bacterial cells of Tepidiphilus thermophilus produced smaller granules and in lower amount.
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Multiple-births in Czech Republic
Dlouhá, Karolína ; Nývlt, Ondřej (advisor) ; Fabiánová, Jana (referee)
The main goal of this thesis is the analysis of multiple-births in Czech Republic between the years of 1950 and 2005 including the international comparison and assess of the influence of assisted reproduction on occurrence of multiple-births. The thesis is divided into five different parts. The first part is dedicated to the analysis of fertility according to a succession of births in Czech Republic while using the indicators of total fertility, the average age of mothers in the time of childbirth and the total number of new-born babies. The second part focuses on multiple-births considering both historical and contemporary point of view and on possible risks for both the mother and the foetus. Next part describes the assisted reproduction, its history, present and commonly used methods. The fourth part is dedicated to the influence of assisted reproduction on the occurrence of multiple-births and to the analysis of multiple-births with the use of absolute and relative number of mono-births and multiple-births indicators, indicators of total fertility, masculinity index, the rate of stillborn babies and the proportion of children born outside of marriage. The last part compares the development of multiple-births within the European countries according to NIDI 1999 classification.
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Freedom at work
Dlouhá, Karolína ; Palíšková, Marcela (advisor) ; Legnerová, Kateřina (referee)
This Master´s Thesis is concerned with freedom at work. The aim of this thesis, is describe, analyse and evaluate the concept "Freedom at work" in three particular companies. The thesis is divided into two parts, theoretical and practical. The theoretical part defines the theoretical basis, principles and methods related to this area. Data are largely based on the academic literature. The practical part represents best practices of liberal companies abroad. In this part selected companies Etnetera a.s., IReSoft, s.r.o. and Country Life, s.r.o. are presented. There is also analysis of democratic principles in these companies, survey, survey evaluation and formulation of recommendations. To achieve the goals of this work are used these methods: comparison of theory with practice, interviews with the members of the personnel department and the director, interviews with employees of liberal companies, analysis of internal documents and questionnaires. The acquisition of this work is a formulation of recommendations that emerged from the comments and suggestions of employees in the questionnaire survey.
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Territorial diferenciance fertility in the Czech Republic
Dlouhá, Karolína ; Langhamrová, Jitka (advisor) ; Cséfalvaiová, Kornélia (referee)
Abstract The main purpose of this following report is to describe and compare the development of fertility and abortions indicators in a particular time and than evaluate regional differences of these indicators in all the regions of Czech Republic. These regions will be than deliminated into several groups based on the similarity of the developement of fertility and abortions. The first part of this report is mainly focused on explanation of basic terms, delimination of indicators and description of the methods of adjusting these indicators. Second part describes the developement of aggregate fertility and abortions indicators and also specific levels of fertility and abortions since the beginning of 1990's. Third part is dedicated to delimination of Czech Republic regions after the NUTS classification and to the following analysis of fertility and abortions indicators in several regions of Czech Republic. These regions are than deliminated into several groups according the similarity of developments.
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