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Application of thermophilic Culture for Organic Compounds Biodegradation
Piechová, Jana ; Burdychová, Radka (referee) ; Babák, Libor (advisor)
The aim of this work is focused on biodegradability study of modified polyurethane materials (foams and foils) in glukose medium and biodegradation of whey. The biodegradability was tested with using mixed thermophilic aerobic bacterial culture of the Bacillus and Thermus genera. The cultivations were practised in the fermentor and in Erlenmayer flasks in a heated shaker. Maximal reduction of COD and culture growth has been achieved by biodegradation of polyuretane in the fermentor. Main effect of degradation was abiotic destruction of PUR materials and consequent utilization of degradation products by bacterial culture. These tests indicated that the best modification agent for biodegradation was 10% carboxymethyl cellulose. The polyurethane foam was more decomposed then the polyurethane foil. Biodegradation of whey without proteins in the fermentor was detected decrease of CHSK and lactose concentration about 50 % in 20th hour. Optimal conditions for whey biodegradation with the thermophilic bacterial culture were pH 6,5 and temperature 60 °C.
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Use of Kluyveromyces marxianus to bioethanol produce from waste paper
Tomečková, Andrea ; Hrstka, Miroslav (referee) ; Babák, Libor (advisor)
The diploma thesis is focused on production possibilities of bioethanol from waste paper by yeast Kluyveromyces marxianus. Waste cardboard was used as a potential substrate for bioethanol production. Several methods for cardboard preparation were introduced and compared as well as methods of fermentation. Simultaneous sacharification and fermentation and separate hydrolysis and fermentation of preprepared cardboard paper were performed in different pH buffer (4,8-7). Simultaneous sacharification and fermentation was held at a temperature of 45°C. Hydrolysis in separate hydrolysis and fermentation was performed at 50°C and fermentation at 25°C. Procedures outputs were obtained by sampling in specific time intervals and samples were analyzed by HPLC for presence and concentration glucose and ethanol. The results of the analysis have shown that the highest concentration of glucose produced by enzymatic hydrolysis was achieved by using microwaves, 2% H2SO4 and 2% NaOH pretreated paperboard at pH 4,8. The highest yield of ethanol was obtained by separate hydrolysis and fermentation of pulp pretreated by microwaves, 2% H2SO4 and 2% NaOH in pH 5,4 buffer. The method SHF proved to be more effective for the production of ethanol than SSF.
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Study of aerobic cultivation conditions with select strain of Lactobacillus
Šupinová, Petra ; Burdychová, Radka (referee) ; Babák, Libor (advisor)
The aim of this study was focused on the study of conditions of growth of strains Lbc. paracasei subsp. paracasei CCDM 211, Lbc. paracasei CCDM 212, Lbc. paracasei subsp. paracasei CCDM 213 and Lbc. salivarius CCDM 216 in media with different amount of carbon-source (glucose, lactose and whey). Next part of the experiment was dealed with study of conditions of bacteria growth at stress conditions (lower pH). The purity od bacterial culture was verified with help of streaking. Purity DNA isolated from bacteria was tested using agarose gel electrophoresis, DNA concentration was estimated spectrophotometricaly. The presence of bacteria of genus Lactobacillus was proved using polymerase chain reaction (PCR) with genus specific primers.
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Whey utilization with thermophilic microorganisms
Rychová, Alexandra ; Vítová, Eva (referee) ; Babák, Libor (advisor)
This diploma thesis studies the utilization of whey using thermophilic bacteria of the genus Thermus and Geobacillus. The whey stripped off proteins was used as a cultivation medium during experiments. The cultivation took place in the Erlenmeyer flasks, to assess the optimal conditions for microrganism’s growth. During the cultivation in the bioreactor, growth curves were established. The amount of whey utilization was assessed by analytical methods that determine the concentration of reducing saccharides (lactose) and chemical oxygen demand (COD) while studying the optimal conditions and a method determining the concentration of reducing saccharides during growth curves analysis.
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Mechanically separated meat from poultry produced by different techniques: Proteins, fat and bone fragment determination.
Čížková, Silvie ; Babák, Libor (referee) ; Šalplachta, Jan (advisor)
The diploma thesis is focused on mechanically separated chicken meat (MSM), which was produced on the machine Baader 601 (soft way of separation) and Lima D (hard way of separation). The history, production, composition and legislation relating to the MSM have been described in the theoretical part. The experiments involving the results of the determination of total solids, total fat, total protein, muscle protein and collagen have been introduced in the experimental part. Bone fragments have been determined by weight and visually after alkaline hydrolysis of samples at elevated, respectively atmospheric pressure. Results have been evaluated with regard to the feedstock and the method of mechanical separation. Values have been compared with data published in the scientific literature and legislative requirements. Data from the experiment confirm high variability and thus different food quality of MSMs, depending on the sort of feedstock used in the production of MSM and the separation method.
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Study of polyhydroxybutyrate production in bacteria
Melušová, Soňa ; Babák, Libor (referee) ; Márová, Ivana (advisor)
Presented work is focused on study of polyhydroxybutyrate production in bacteria. In theoretical part short characterization of PHB was given and the most common representative of wide group of polyhydroxyalkanoates (PHA) were described. Then, production of PHB and copolymer P(HB-co-HV) in selected bacterial strains was experimentally proven. First, PHB production in Bacillus megaterium using synthetic medium was studied. The PHB content in cells was increased during cultivation under limiting conditions, despite low growth. Addition of ethanol into production media resulted in increased PHB synthesis as well as biomass production (21 % PHB of 1,8 g/l biomass). Further, BM medium containing 8 g/l glucose was tested. PHB production was more than 1 g/l at significant growth increase when compared with synthetic medium. The bacteria B.megaterium showed, except glucose, ability to utilize maltose and xylose. Another cultivations were tested with bacterial strain Azotobacter vinelandii, which is capable of copolymer P(HB-co-HV) synthesis. Maximal growth and copolymer content was reached on Burk's medium with 30 g/l of glucose. Addition of peroxide to growth medium influenced P(HB-co-HV) synthesis to 46 % of 2,6 g/l biomass. Bacteria A.vinelandii showed the best growth on maltose, even compared with glucose (54 % copolymer of biomass content). Finally, PHB production on industrial waste product – whey was monitored. Using Plackett-Burman design for statistical media optimization, the whey content was modified. B.megaterium grown on adjusted whey reached 0,5 g/l PHB, 32 % of cell's content.
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Study of growth and optimization of selected metabolites production by Zymomonas mobilis
Lukačková, Adéla ; Vránová, Dana (referee) ; Babák, Libor (advisor)
In the diploma thesis are discussed the process of enzymatic hydrolysis of waste paper as a source for the production of bioethanol by bacteria Zymomonas mobilis. In the theoretical part summarize basic information about particular methods of hydrolysis, about paper used as a raw material for enzymatic hydrolysis, about possibilities of the fermentative production of bioethanol focusing on the method of simultaneous saccharification and fermentation comparison with enzymatic hydrolysis and fermentation. Suitable microorganisms for ethanolic fermentation and simultaneous saccharification and fermentation and their advantages and disadvantages, are further discussed in this part as well. The theoretical part ends with the suggestion of the technological process for production of bioetanol. It covers all necessary steps from the input of raw material to the separation of produced ethanol. In the experimental part various parameters of hydrolysis, fermentation and simultaneous saccharification and fermentation were optimized using enzymes from Novozymes® company and the Zymomonas mobilis CCM2770 and Zymomonas mobilis LMG457 bacterium. The conversion rate of paper cellulose to gluckose and production of ethanol were observed by HPLC/RI method. Type of buffer, quantity of cells, enzyme and substrate were optimized in order to maximize the efficiency of the process. All experiments were performed on paper containing high amount of cellulose and for comparison on standard medium which contains gluckose. The highest yields was achieved with the use of Novozymes® Cellulosic ethanol enzyme Kit. The strain Zymomonas mobilis LMG457 has demonstrated as a better producer.
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Changes of fatty acids during production of model cheese
Janoušková, Kateřina ; Babák, Libor (referee) ; Vítová, Eva (advisor)
This thesis deals with the identification and quantification of free and bound fatty acids in model cheese samples of the Moravský bochník type, which were produced at Tomas Bata University in Zlín by a standard technological procedure. In a theoretical part of the thesis, the issues of lipids, fatty acids, as well as the characteristics of high cooked cheeses including Moravský bochník, microbial cultures used in their production, and the possibilities of fatty acids determination are summarized. The ČSN EN ISO 1735:2005 method was chosen for the lipid extraction from cheese samples. Present free and bound fatty acids were identified and quantified by gas chromatography with flame-ionization detection after conversion to methylesters by acid esterification with bortrifluoride as a catalyst. In total 32 fatty acids were identified in cheese samples, the highest amount of caproic, caprylic, capric, lauric, myristic, palmitic, stearic and oleic acids.
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Waste materials utilization for preparing hydrolysates for the fermentation phase.
Vadovičová, Natália ; Hrstka, Miroslav (referee) ; Babák, Libor (advisor)
Bachelor thesis focuses on the study and comparison of different types of hydrolysis, their optimization and maximization of yields for the upcoming fermentation. Orange peel was chosen as a substrate to conduct the experiments. First, the substrate was mechanically grinded to form a suspension. Each suspension then underwent one out of the examined methods of hydrolysis. Chosen methods were physical, such as microwaves, increased temperature or ultrasound, and chemical acidic and alkaline. Combinations of both types were also examined. The last optimized method was enzymatic hydrolysis. First set of experiments was conducted using enzymes Novozymes® NS50013 and NS50010. Production of cellulase and pectinase enzymes by A. niger during solid-state fermentation that lasted 10 days was also studied. The yields of reducing sugars of all the experiments were calculated using the Somogyi-Nelson method. Enzymatic hydrolysis was proven to be the most effective using the combination of both of the enzymes for a period of 96 hours at pH = 4.5 and temperature 45 °C. Yield of the reducing sugars under these conditions reached 27,4241 ± 0,0007 gl-1.
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Thermotolerant strains application for preparation of selected metabolites on waste hydrolysates
Musilová, Kristína ; Vítová, Eva (referee) ; Babák, Libor (advisor)
This Bachelor´s Thesis deals with the preparation of the selected metabolite on hydrolysates of orange fruit waste, specifically on orange peel hydrolysates using thermotolerant strains. The thermotolerant Bacillus coagulans strain having lactic acid as the primary metabolite, a substance with a great perspective, was selected for the fermentation process for its exceptional properties. At the beginning of the experiment, optimal conditions for the growth of the selected microorganism were determined; these were applied during the fermentations. Several fermentations were performed under various conditions until those conditions were determined, in which lactic acid was produced. The fermentation production was determined by the HPLC method and the saccharide decrease by the Somogyi-Nelson method. To achieve lactic acid production, it was necessary to extract the orange peels by an organic agent before fermentation, in order to eliminate substances causing inhibition of micro-organism growth. Higher yields were achieved by the SHF method, but the highest by using the Lactobacillus casei strain.
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