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Utilisation of plant origin waste
Habáníková, Kamila ; Šťavíková, Lenka (referee) ; Omelková, Jiřina (advisor)
This thesis describes properties and utilization and specifies the composition of organic plant waste produced during industrial processing. By cultivation of specific micro-organisms from this waste material it is possible to produce enzymes that have further beneficial industrial utilization. The experimental part of this thesis deals with the production of enzymes from the applied waste material grape pomace with the help of chosen micro-organisms. Waste, its processing and utilization is and always will be a key issue in our society.
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Production of PHB-HV from grape pomace by employing a pure culture of Cupriavidus necator
Jakešová, Michaela ; Kučera, Dan (referee) ; Kovalčík, Adriána (advisor)
This bachelor thesis deal with the production of PHB-HV copolymer employing a pure culture of Cupriavidus necator H16. First, Erlenmeyer flask´s cultivations using pure fructose as a source of carbon were performed. The mode of addition of valeric acid for the synthesis of PHB-HV was studied. Furthermore, the effect of stress factor (ethanol) on PHB-HV synthesis by this microorganism was investigated. The method obtained by optimization in Erlenmeyer flasks was applied to the synthesis of the PHB-HV copolymer in the bioreactor using fructose as a source of carbon, too. Furthermore, after enzymatic hydrolysis, a sugar extract from grape pomace was obtained as a source of carbon for the synthesis of PHB-HV. The fermentation in Erlenmeyer flasks and the bioreactor using an extract of grape pomace was carried out. Using this alternative carbon source 3.51 g/l PHB-HV copolymer was produced in Erlenmeyer flask, 5.06 g/l in bioreactor respectively. In the first case, the proportion of 3-hydroxyvalerate in the copolymer was 23.78 mol. % and in the second one, 9.39 mol. %.
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Employment of bacterium Tepidimonas taiwanensis for production of polyhydroxyalkanoates on waste substrates
Vidláková, Michaela ; Pernicová, Iva (referee) ; Obruča, Stanislav (advisor)
This diploma thesis is focused on screening PHA production using thermophilic bacterium Tepidimonas taiwanensis and on the study of possible use of grape pomace, molasses and waste paper as a cheap carbon substrate for culturing the characterized bacteria. At first, testing of basic cultivation parameters was performed, including carbon substrate concentration, oxygen availability, ability to utilize nitrogen sources and selected disaccharides. PHA production from waste substrates was tested in three ways. In the first, pre-prepared solids-free hydrolysates from raw materials were used as the carbon source. The second and third procedures were performed by dosing waste materials directly into the mineral media, which differed only in the presence or absence of the enzyme preparation enabling release of fermentable sugars. The most intensive increase in culture and the highest production of PHA was recorded on grape hydrolyzate. The biomass concentration in this sample reached up to 4.8 g/L with a content of 59 % PHA. On the other hand, the addition of grape marc directly to the production medium did not work at all, which was probably due to the presence of a large amount of inhibitory substances from the pomace. The situation was similar with molasses and waste paper, where the bacterium Tepidimonas taiwanensis was able to grow and possibly produce PHA only to a small extent. The work also managed to characterize the effect of temperature and pH on the activity of the enzyme cocktail Viscozyme L and to determine the temperature and pH optimum PHA synthase of the bacterial strain Tepidimonas taiwanensis in the cell lysate.
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Use of secondary raw materials from wine production
Lajtman, Roman ; Vespalcová, Milena (referee) ; Diviš, Pavel (advisor)
This bachelor thesis deals with using of grape pomace, waste from wine industry. It also deals with the determining of total phenolic substances in a grape pomace. The theoretical part focuses on the characteristic of grape, the chemical composition of wine, making of wine and mainly on use of secondary raw materials. It also describes phenolic substances, their division and finally something general about using experimental methods. In the experimental part a red grape pomace´s extraction was made. The extractions were made in different solvents, in a mixture ethanol-water (ratio 20:80; 40:60 and 60:40), also in a mixture glycerol-water (ratio 10:90; 20:80 and 40:60) and at the end in water. The extractions were made in a different time and the purpose was to optimize the extractions and to get the highest yield of phenolic substances, antioxidants. Time and economically side played an important role to find out and to choose the best solvent. In the results, as the best and the optimal extraction´s solvent on extraction of red grape pomace was chosen the mixture ethanol-water and ratio was 60:40. That mixture extracted the most phenolic substances and relatively in the shortest time. During the optimization of sample weight was found out that optimal sample weight is 4 g of grape pomace. At the end repeated extraction was made. It is obviously practically worthless. After second step of extraction, the concentration of phenolic substances is more than a half lower than at the first step.
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Prospects of Microbial Degradation and Waste Utilization from Food Processing Industry
Illková, Kateřina ; Karovičová, Jolana (referee) ; Jarošová, Alžběta (referee) ; Omelková, Jiřina (advisor)
This work deals with the problem of microbial degradation of the waste materials from food industry. This work is focused on the production of technological significant enzymes producing by microorganisms, which were able to use the waste as a sole carbon source. In the first part of this work, the attention was focused on the production of pectolytic enzymes. This part was made within study interships in Slovak Academy of Sciences, department of Glycomics in Bratislava. The grape pomace as the waste form winegrowing was used as a sole carbon source for microbial growth and enzymes production. The production of pectolytic enzymes was tested on this waste. After screening the most suitable microorganisms was chosen with the highest production of polygalacturonase activity. Produced enzymes were isolated by extraction techniques, purified and then identified proteomically. The aim of the second part of this work was the waste water treatment containing lipids and lipolytic enzymes. The reason was the cooperation with the company constructing grease traps. The characterization of supplied commercial preparations was the subject of this work and the other reason was the characterization of conditions for lipases secreting by microorganisms, identification of microorganisms present in the commercial preparation and testing of new microbial cultures for the development of new preparation for the grease traps.
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Partial purification and characterization of polygalacturonases of Geotrichum candidum.
Jäger, Jakub ; Breierová, Emília (referee) ; Stratilová, Eva (advisor)
This work discusses the possibilities of using microbial degradation of grape pomace, main waste material from wine production, to preparate industrially important enzymes. The issue is focused on the production of pectolytic enzymes, particularly polygalacturonase, by Geotrichum candidum CCY 16-1-29 via solid state fermentation on grape pomace. The theoretical part of the bachelor thesis focuses on studying plant cells and saccharides from which the plant cell wall is made of, mainly pectin. Cell wall sacharides were used as a carbon source for solid state fermentation (SSF) and pectin as an inductor of pectolytic enzymes. This bachelor thesis also deals with the enzymatic degradation of cell wall polysacharides. The greatest attention is paid to degrade pectin and pectolytic enzyme function. Production of pectolytic enzymes is mentioned subsequently. The last chapter from the theoretical part is dedicated to technical use of pectolytic enzymes. In the experimental part of this work I deal with the partial purification and characterization of majority polygalacturonase produced on the seventh day of cultivation, when another increase of extracellular polygalacturonase activity occurred. The yield of cultivation was 43,5 mg of protein extract /100 g of grape pomace. The extract contained protein, and its activity was lyophilisate. Its specific activity was protein. The enzyme was produced in at least four forms differing in pH optimum (4,0; 4,4; 4,8; 5,2). The pH optimum for majority polygalacturonase was 4,8. Action pattern of this enzyme determined as the dependence of polymeric substrate viscosity decrease on its degradation showed that the enzyme is a typical polygalacturonase with random action pattern (EC 3.2.1.15).Value of Km reached indicating a high affinity for this substrate. The amino acid sequence "SNNVVSNVNILSSQVVNSDNGVR" obtained by mass spectrometry after SDS-PAGE and tryptic digestion, was identified as a stretch of primary structure of polygalacturonase of Ap2PG1 G. candidum based on the comparison with proteins from the Uniprot database. It shows the highest similarity with other polygalacturonases of G. candidum S31PG1, S31PG2 and G. klebahnii PSE3. On the basis of this similarity to enzymes produced by phytopathogenic strains of G. candidum and the fact that this enzyme was not produced only in the early stages of cultivation, it can be assumed, that the strain of G. candidum CCY 16-1-29 acted also as a phytopathogenic strain.
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Use of wastes from winemaking in food and cosmetics applications
Výmolová, Markéta ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The presented thesis deals with the various types of hydrolysis of waste from the production of wine, parings and bunch-stems for subsequent biotechnological use as enrichment of feeding mixtures and the production of bio-ethanol. A composite analysis was done at the start of the measurements for the determination of dry matter, extraction of oil from the parings and assessment of cellulose in the bunch-stems. As a part of this thesis a chemical and enzyme hydrolysis of waste materials have been performed. In hydrolysates the contents of reducing carbohydrates and antioxidants, polyphenols and flavonoids were analysed. The results showed that the samples with a concentration of 50 g/l of material hydrolysed in a water bath for 60 minutes were the best conditions for cultivating yeasts. Hydrolysed parings and bunch-stems were used as the production medium for the production of ethanol in Saccharomyces cerevisiae. A change of the concentration of carbohydrates and ethanol in the samples has been measured by HPLC. The sample hydrolysed enzymatically using cellulase exhibited the highest concentration of biomass (c = 2,140 g/l), the sample hydrolysed chemically in the water bath formed the highest content of ethanol (c = 9,880 g/l).
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Preparation and characterization of microcrystalline cellulose from food waste
Horáková, Natálie ; Němcová, Andrea (referee) ; Kovalčík, Adriána (advisor)
The aim of the diploma thesis was to isolate microcrystalline cellulose from grape pomace. The analysis of grape pomace showed the following main components: lignin (32.9%), cellulose (34.2%), hemicellulose (31.5%) and lipids (7.0%). Various methods have been tested for the isolation and purification of microcrystalline cellulose. As the most suitable method was selected acid hydrolysis with 50 % and 64 % sulfuric acid followed by treatment with ethylenediamine. The morphology of the isolated cellulose was studied by transmission electron microscopy. The diameter of the hydrolyzed cellulose fibers was determined in the range of 15–130 nm. The isolated cellulose was used as reinforcing filler for agar. The agar/cellulose films were prepared by solution casting. The agar/cellulose films exhibited higher values of Young´s moduli and tensile strength but lower elongation at break compared to neat agar films. The results proved that grape pomace can be used as source for the isolation of microcrystalline cellulose with reinforcing ability for agar.
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Biotechnological production of PHA from waste substrates
Krejčiřík, Roman ; Hrabalová, Vendula (referee) ; Obruča, Stanislav (advisor)
The presented bachelor thesis deals with the study of polyhydroxyalkanoates using wine grape pomace extract. The thesis aims to determine whether it is possible to use wine grape pomace extract as a substrate for polyhydroxyalkanoates (PHA) production. For this research, four thermophilic bacterial strains were selected such as Schlegelella thermodepolymerans, Caldimonas meghalayensis, natural isolate labeled as 34, and Tepidimonas taiwanensis, from which the most suitable bacterium was subsequently selected. The best transformation of residual sugars showed T. taiwanensis, and therefore it was used for all the further fermentation. The extracts were prepared out of wine marc of Veltlínské červené rané, Müller Thurgau, Pálava, Sauvignon, Rulandské bílé, Frankovka (rosé), Frankovka (red wine). The highest PHA concentration was detected after fermentation with concentrated extracts of Sauvignon and Rulandské bílé. Fermentation processes in other extracts were most likely inhibited by the components they contained. Optimization was performed by dilution of extracts with water. The highest concentrations of PHA while using dilute extracts were achieved with the extract from different kinds of Frankovka (rosé) and Frankovka (red wine).
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Employment of bacterium Tepidimonas taiwanensis for production of polyhydroxyalkanoates on waste substrates
Vidláková, Michaela ; Pernicová, Iva (referee) ; Obruča, Stanislav (advisor)
This diploma thesis is focused on screening PHA production using thermophilic bacterium Tepidimonas taiwanensis and on the study of possible use of grape pomace, molasses and waste paper as a cheap carbon substrate for culturing the characterized bacteria. At first, testing of basic cultivation parameters was performed, including carbon substrate concentration, oxygen availability, ability to utilize nitrogen sources and selected disaccharides. PHA production from waste substrates was tested in three ways. In the first, pre-prepared solids-free hydrolysates from raw materials were used as the carbon source. The second and third procedures were performed by dosing waste materials directly into the mineral media, which differed only in the presence or absence of the enzyme preparation enabling release of fermentable sugars. The most intensive increase in culture and the highest production of PHA was recorded on grape hydrolyzate. The biomass concentration in this sample reached up to 4.8 g/L with a content of 59 % PHA. On the other hand, the addition of grape marc directly to the production medium did not work at all, which was probably due to the presence of a large amount of inhibitory substances from the pomace. The situation was similar with molasses and waste paper, where the bacterium Tepidimonas taiwanensis was able to grow and possibly produce PHA only to a small extent. The work also managed to characterize the effect of temperature and pH on the activity of the enzyme cocktail Viscozyme L and to determine the temperature and pH optimum PHA synthase of the bacterial strain Tepidimonas taiwanensis in the cell lysate.
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