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Methods for identification of PHA producing bacteria
Skřivanová, Veronika ; Turková, Kristýna (referee) ; Obruča, Stanislav (advisor)
This diploma thesis deals with testing, optimazing and comparing methods for the identification of bacteria producing polyhydroxyalkanoates. Work included cultivation and microscopy methods, wherein the bacterial cells were stained with lipophilic dyes Nile red and Sudan black. Further, we also used flow cytometry and spectroscopic methods - Raman spectroscopy and infrared spectroscopy with Fourier transformation, and molecular biological methods, which analyzed the presence of a gene encoding PHA synthase (phaC) by polymerase chain reaction (PCR). PCR assay consist of two reactions, the firt on eis based on amplification of phaC gene along with 16S rRNDA gene, which is common for all the bacteria (multiplex PCR). The second reaction is focused on specific amplification of PHA synthase catalyzing biosynthesis of mcl-PHA. In order to overcome false positive results typical for methods analyzing genotype and also to avoid false negative results occuring in fenotype analyzing methods, the best strategy is to combine both aproaches. According to our results, analysis of presence of phaC gene by PCR can be combined with methods capable of determining presence of PHA in bacterial cells. For this purpose, Raman microspectroscopy seems to be very promising tool, since it is able to detect low content of PHA in cells and PHA can not be confused with other lipid metabolites. The results provide an overview of test methods, their advantages and disadvantages and also to compare different criteria according to which it is possible to choose the method of identification in depending on the adjustable requirements.
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Everyday life in Slaný around year 1600
Hlušičková, Jana ; Čechura, Jaroslav (advisor) ; Skřivanová, Veronika (referee)
Presented thesis deals with the everyday life of the 16th and 17th century in the royal town of Slaný in the central Bohemia. It recounts the eventful history of the city from its prehistoric beginnings to the present. Focus is the fourth and fifth chapter. The fourth chapter discusses the type sof criminal offenses and punishment for them in the early modern period grant. This includes offenses against life and healthy, against property, against morality and honor. This section we also learn about dishonest professions. The fifth chapter describes the everyday concerns of people at the time - the need to belong, cocern for the livelihood, concern for subsistence but also financial matters. Full text concludes humorous incident that happened in 1604.
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Everyday life in Slaný around year 1600
Hlušičková, Jana ; Čechura, Jaroslav (advisor) ; Skřivanová, Veronika (referee)
Presented thesis deals with the everyday life of the 16th and 17th century in the royal town of Slaný in the central Bohemia. It recounts the eventful history of the city from its prehistoric beginnings to the present. Focus is the fourth and fifth chapter. The fourth chapter discusses the type sof criminal offenses and punishment for them in the early modern period grant. This includes offenses against life and healthy, against property, against morality and honor. This section we also learn about dishonest professions. The fifth chapter describes the everyday concerns of people at the time - the need to belong, cocern for the livelihood, concern for subsistence but also financial matters. Full text concludes humorous incident that happened in 1604.
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Correspondence of Kutná Hora and Kolín from 1560s to White mountain
Skřivanová, Veronika ; Vaněk, Vojtěch (referee) ; Čechura, Jaroslav (advisor)
The final thesis investigates 801 traced inscriptions which originate in four city books of received correspondence of Kutná Hora and in ten city books of received correspondecne of Kolín. Further sources are 2 letteres from the years 1587 and 1588 from the State archive of the district of Kolín and 2 letteres from the years 1573 and 1587 from the State archive of the district of Kutná Hora. In the introduction, there is mentioned the summary of the literature which deals with city`s, noble`s and sovereign`s correspondence. The terms "city book of sent correspondence" and "city book of received correspondence" are explained and the characterization of used sources occurs as well. This work explains to us the environment of the town of Kutná Hora and Kolín in the Pre -White Mountain Period. The 4th chapter of this work deals with the term and the process of communication not only theoratically, but first of all practically on the example of Kutná Hora`s and Kolín`s city counsellors. The 5th chapter is focused on the content of official correspondence sent between Kutná Hora and Kolín. Through the citations we will learn more about the property matters of dwellers of both cities. We will mention cases concern city corporations and matters of commune character. In the annexe to this final thesis can be found...
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Methods for identification of PHA producing bacteria
Skřivanová, Veronika ; Turková, Kristýna (referee) ; Obruča, Stanislav (advisor)
This diploma thesis deals with testing, optimazing and comparing methods for the identification of bacteria producing polyhydroxyalkanoates. Work included cultivation and microscopy methods, wherein the bacterial cells were stained with lipophilic dyes Nile red and Sudan black. Further, we also used flow cytometry and spectroscopic methods - Raman spectroscopy and infrared spectroscopy with Fourier transformation, and molecular biological methods, which analyzed the presence of a gene encoding PHA synthase (phaC) by polymerase chain reaction (PCR). PCR assay consist of two reactions, the firt on eis based on amplification of phaC gene along with 16S rRNDA gene, which is common for all the bacteria (multiplex PCR). The second reaction is focused on specific amplification of PHA synthase catalyzing biosynthesis of mcl-PHA. In order to overcome false positive results typical for methods analyzing genotype and also to avoid false negative results occuring in fenotype analyzing methods, the best strategy is to combine both aproaches. According to our results, analysis of presence of phaC gene by PCR can be combined with methods capable of determining presence of PHA in bacterial cells. For this purpose, Raman microspectroscopy seems to be very promising tool, since it is able to detect low content of PHA in cells and PHA can not be confused with other lipid metabolites. The results provide an overview of test methods, their advantages and disadvantages and also to compare different criteria according to which it is possible to choose the method of identification in depending on the adjustable requirements.
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Hydrolytic cleavage of coffee spent ground
Skřivanová, Veronika ; Hlaváček, Viliam (referee) ; Obruča, Stanislav (advisor)
The bachelor thesis deals with various types of hydrolysis of spent coffee ground for the following biotechnological applications such as the culture medium. The work included mechanical, acid, alkaline and enzymatic hydrolysis. The resulting hydrolysates were analyzed concerning the content of reducing carbohydrates, and potentially inhibitory substances polyphenolic nature. The results show that the best conditions for culturing yeast samples were prepared with a concentration of coffee 150 g/l hydrolysed combination of 2,5% mineral acid by heating at 121 °C for 15 minutes. Using ultrasonic homogenizer significantly increases the effect of hydrolysis. The enzymatic hydrolysis is best placed hydrolyzate prepared cellulase. The hydrolysed spent coffee grounds were used as the production medium for the production of ethanol by Saccharomyces cerevisiae. For samples was determined concentration change of glucose and ethanol by HPLC/RID. The highest concentration of biomass sample contained hydrolyzed cellulose on the other side, the highest yields of ethanol (20,07 g/l) were achieve using hydrolysate prepared by 2,5% hydrochloric acid.
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