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Microbial lipases and their application
Pavlačková, Jana ; Flodrová, Dana (referee) ; Omelková, Jiřina (advisor)
This diploma thesis is focused on the study of the preparation for fat separators and wastewater pipes that contains the microorganisms with lipolytic activity. Theoretical part of this thesis describes lipases, microorganisms producing this enzymes and usage of lipases. In this part possibilities of identification of microorganisms are presented too. The practical part is concerned with the study of commercial preparation Sany Duo Spezial with proven presence of microorganisms with lipolytic activity. These microorganisms were identified by means of the PCR method. This method identified mictoorganisms like genus Bacillus sp. Next characterization of the preparation was focused on the determination of COD and the investigation of the influence of various conditions of culture medium on the lipases production and their activity. The effect of temperature, ions and pH was studied. Lipolytic activity was determine spectrophotometricaly with usage of p-nitrophenyllaurate whitch dissociates to yellow product p-nitrophenol.
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Utilization of biomass for the energy purposes
Romanová, Kristýna ; Ing.Ondřej Kosík, Ph.D. (referee) ; Flodrová, Dana (advisor)
The theoretical part of work focuses on the issue of biomass used for energy purposes, in particular agricultural waste, which can be used as a substrate for biogas station. Furthermore, examines the composition of polymer substrates, which affects most biogas process. The aim of this work was to measure each sample of enzyme activity of biomass, which is used as a raw material for biogas plants.
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Utilization of plant origin waste
Habáníková, Kamila ; Flodrová, Dana (referee) ; Omelková, Jiřina (advisor)
Production of cellulase and polygalacturonase by Aspergillus niger and Aureobasidium pullulans was studied in submerged (SmF) and solid state fermentation (SSF) systems. Substrates used in fermentation systems were mandarin peels and grape pomace. With Aspergillus niger used on grape pomace as a sole carbon source, cellulase production was detected after 72 hours in SSF and after 24 hours in SmF systems. The activity of cellulase per gram of substrate was higher in a submerged than in a solid state fermentation system. The longer time for higher polygalacturonase production was necessary in submerged fermentation systems and polygalacturonase activity was higher in SmF. The SSF fermentation with mandarin peels as a sole carbon source was similar, the highest detected activity of cellulase was determined after 72 hours. Different production of polygalacturonase was observed on mandarin peels in SmF systems. A comparison of enzyme productivities on grape pomace and mandarin peels showed that polygalacturonase activity per gram of substrate is highest in SmF system with mandarin peels as a sole carbon source. With Aureobasidium pullulans used on grape pomace as a sole carbon source, cellulase production was detected after 48 hours in SmF and SSF fermentation systems. The activity of cellulase per gram of substrate was higher in solid state system than in a submerged fermentation system. Longer time for higher polygalacturonase production was necessary in both fermentation systems. Polygalacturonase activity was higher in SmF. The SSF fermentation with mandarin peels as a sole carbon source was similar, the highest detected activity of cellulase was determined after 48 hours. Different production of polygalacturonase was observed on mandarin peels in SmF systems. A comparison of enzyme productivities on grape pomace and mandarin peels showed that polygalacturonase activity per gram of substrate is highest in SmF system with mandarin peels as a sole carbon source. For both systems and both substrates manganese-dependent peroxidase was detected for the first time. Differences in the enzyme synthesis by Aspergillus niger and Aureobasidium pullulans depend on both the substrates used as well as on the fermentation system.
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Utilization of micropreparative isoelectric focusing in the analysis of biological samples
Vlčková, Zuzana ; Flodrová, Dana (referee) ; Šalplachta, Jiří (advisor)
The aim of this Master Thesis is the optimization and verification of conditions of separation of proteins from biological materials. For the separation a developed technique called micropreparative isoelectric focusing is being used. The theoretical part describes analytical methods of biological materials with a focus on proteins which were used for the optimization of the developing system in general. This part of the Master Thesis also presents this method itself. Furthermore, basic characteristics of other techniques used to control the efficiency of the micropreparative isoelectric focusing are introduced. The experimental part depicts the individual steps of optimization of the micropreparative isoelectric focusing. The ideal separation procedure was firstly found out on the individual standard proteins, secondly on their mixture and lastly the optimised procedure was applied to a real biological sample. The efficiency of the micropreparative isoelectric focusing was verified by SDS-PAGE and MALDI-TOF mass spectrometry.
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The study of production of hydrolytic enzymes for cellulose wastes treatment
Řezáčová, Barbora ; Flodrová, Dana (referee) ; Omelková, Jiřina (advisor)
The study of production of hydrolytic enzymes dealt with the production of cellulase and polygalacturonase by two microbial strains - Aspergillus niger and Aureobasidium pullulans. The enzymes were produced in solid-state fermentation system. The wheat straw and sugar beet pulp were used as a substrate. The substrates were moistened by water, mineral solution or by medium with glucose. The effect of mineral solution and glucose on production of these enzymes were monitored during cultivation. The highest production of polygalacturonase was achieved by Aspergillus niger during cultivation on sugar beet pulp moistened by mineral solution. The highest production of cellulase was achieved by Aspergillus niger during cultivation on wheat straw moistened by medium with glucose.
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The employment of waste paper to microbial production of cellulases
Peterek, Miroslav ; Flodrová, Dana (referee) ; Omelková, Jiřina (advisor)
Study of the employment of waste paper to microbial production of cellulase was carried out using Aspergillus niger cultivation on carbon sources that have been waste office paper and cardboard, humidified by no – carbon medium or distiled water. Cultivation took place in the SSF way in Erlenmeyer flasks and columns. Concentration of extracellular proteins, cellulase and protease activity for selected samples were monitored. It was found that the most advantageous method of cultivation in terms of cellulase activity production is the cultivation in the column washing by no – carbon medium in three day intervals.
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Degradation of fats in wastes
Artýszková, Jana ; Flodrová, Dana (referee) ; Omelková, Jiřina (advisor)
Submitted master’s thesis is focused on the study of possibilities of lipids degradation in particular wastewaters originating in food industry or restaurants. The effort is given to the employment of lipolytic activity presenting microorganisms. In the literature review, wastewater treatment with aim on the sludge management and fat separators are described, as a way how to pre-treat these wastewaters. In this part the enzymes lipases of microbial origin are researched from point of view of their production conditions and possible applications. The experimental part is dedicated to the research of optimization of cultivation conditions for lipases production employing selected microorganisms (Bacillus subtilis, Geobacillus thermodenitrificans, G. thermocatenulatus and mixed bacterial culture Thermus and Bacillus) and a commercial formulation (Sany Duo Spezial). Lipases production and growth of microorganisms are determined spectrofotometrically on various concentrations of lipids. Moreover, employing the solid nutrition medium, the effect of detergents onto the Bacillus subtilis culture was assessed, since detergents are generally abundant in this particular wastewaters. As a conclusion, vide supra mentioned microorganisms were characterized according to their abilities to degrade triacylglycerols.
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Analysis of selected substances contained in the extract from elderberry twigs
Škeřík, Jan ; Flodrová, Dana (referee) ; Šalplachta, Jiří (advisor)
My diploma thesis deals with the optimization and verification of conditions for separation of proteins, which are contained in the twigs of elder (Sambucus nigra L.) using the technique RP-HPLC. The measurements were made with the use of a HPLC system with a UV-VIS detector. Column Zorbax 300SB-C18 300 4,6 x 250 mm and particles the size of 5 microns were used. The theoretical part describes the attributes and usage of elder and especially its proteins. The basic characteristic of the used HPLC technique is introduced. The possibilities of how to identify proteins are described. The practical part demonstrates the individual steps of optimization of the HPLC method applied to the mixture of standard proteins. The application of this method in the real sample is also reported. The final part of the diploma thesis is focused on the comparison of analysed samples taken in different seasons.
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