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Molecular characterization of selected PHA producers
Kubáčková, Eliška ; Brázda, Václav (referee) ; Obruča, Stanislav (advisor)
This diploma thesis focuses on the molecular characterization of selected PHA producers. Within this work, the PHA producing thermophilic isolates originating from the samples of activated sludge and compost were identified and characterized using molecular biological methods. By sequencing the 16S rRNA gene, the thermophilic isolates were identified and taxonomically classified into the Firmicutes bacterial phylum. In these bacterial isolates, the ability to produce PHA at the genotype level was determined by conventional PCR detection of the phaC gene encoding PHA synthase, which is a key enzyme in PHA biosynthesis. Class I, II and IV PHA synthases were detected in most of the isolated bacteria, wherein class I and II PHA synthases are not characteristic for these bacterial genera. The largest proportion of isolates was identified for the species of thermophilic bacterium Aneurinibacillus thermoaerophilus, in which class IV PHA synthase was detected. In the second part of the diploma thesis, the RT-qPCR method was implemented to study the expression of selected genes of the bacterium Cupriavidus necator H16 involved in PHA metabolism. As part of the implementation of this method, PCR-based detection of selected genes was optimized and quantification of genes using real-time PCR was performed. The tested method included steps of RNA isolation, cDNA synthesis and quantification of gene segments for which the critical points of the method were determined based on the obtained data.
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Active components of technical cannabis and their application in cosmetics
Kubáčková, Eliška ; Bokrová, Jitka (referee) ; Němcová, Andrea (advisor)
Hemp is unique plant with many therapeutic effects thanks to its large content of substances. This bachelor thesis brings information about characterization of main biological active substances in cannabis sativa, which are described in theoretical part of this thesis and determined in experimental part of this thesis. As a main active substance were determined antioxidants and cannabinoids for which the liquid chromatography method was optimized. This bachelor thesis also describes the antimicrobial effect of active cannabis substances for which it is used in the cosmetic industry. This antimicrobial activity has been studied in terms of the effect against dandruff, which is primarily caused by the high incidence of Malassezia yeasts on the skin of human hair. For the experimental part, the extracts of the technical hemp and more specific varieties of Bialobrzeskie and Finola were tested, which were further used for the preparation of the anti-dandruff shampoo. By analyzing the active substances in hemp extracts, the uniform result was obtained - the highest content of antioxidants and cannabinoids was reached by preparing of extract from dried hemp flowers. In the last part of the thesis, the antimicrobial effect of hemp extracts and prepared hemp shampoos was determined on selected strains of microorganisms and it was observed antimicrobial activity in extracts and shampoos againts Malassezia furfur yeast.
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Molecular characterization of selected PHA producers
Kubáčková, Eliška ; Brázda, Václav (referee) ; Obruča, Stanislav (advisor)
This diploma thesis focuses on the molecular characterization of selected PHA producers. Within this work, the PHA producing thermophilic isolates originating from the samples of activated sludge and compost were identified and characterized using molecular biological methods. By sequencing the 16S rRNA gene, the thermophilic isolates were identified and taxonomically classified into the Firmicutes bacterial phylum. In these bacterial isolates, the ability to produce PHA at the genotype level was determined by conventional PCR detection of the phaC gene encoding PHA synthase, which is a key enzyme in PHA biosynthesis. Class I, II and IV PHA synthases were detected in most of the isolated bacteria, wherein class I and II PHA synthases are not characteristic for these bacterial genera. The largest proportion of isolates was identified for the species of thermophilic bacterium Aneurinibacillus thermoaerophilus, in which class IV PHA synthase was detected. In the second part of the diploma thesis, the RT-qPCR method was implemented to study the expression of selected genes of the bacterium Cupriavidus necator H16 involved in PHA metabolism. As part of the implementation of this method, PCR-based detection of selected genes was optimized and quantification of genes using real-time PCR was performed. The tested method included steps of RNA isolation, cDNA synthesis and quantification of gene segments for which the critical points of the method were determined based on the obtained data.
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Active components of technical cannabis and their application in cosmetics
Kubáčková, Eliška ; Bokrová, Jitka (referee) ; Němcová, Andrea (advisor)
Hemp is unique plant with many therapeutic effects thanks to its large content of substances. This bachelor thesis brings information about characterization of main biological active substances in cannabis sativa, which are described in theoretical part of this thesis and determined in experimental part of this thesis. As a main active substance were determined antioxidants and cannabinoids for which the liquid chromatography method was optimized. This bachelor thesis also describes the antimicrobial effect of active cannabis substances for which it is used in the cosmetic industry. This antimicrobial activity has been studied in terms of the effect against dandruff, which is primarily caused by the high incidence of Malassezia yeasts on the skin of human hair. For the experimental part, the extracts of the technical hemp and more specific varieties of Bialobrzeskie and Finola were tested, which were further used for the preparation of the anti-dandruff shampoo. By analyzing the active substances in hemp extracts, the uniform result was obtained - the highest content of antioxidants and cannabinoids was reached by preparing of extract from dried hemp flowers. In the last part of the thesis, the antimicrobial effect of hemp extracts and prepared hemp shampoos was determined on selected strains of microorganisms and it was observed antimicrobial activity in extracts and shampoos againts Malassezia furfur yeast.
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