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Production of extracellular hydrolases by specific mould species
Pokrývková, Zuzana ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
This bachelor's thesis is focused on studying the possibilities of producing extracellular hydrolytic enzymes by the fungus Penicillium lilacinum on two substrates. The theoretical part is focused on the characterization of selected hydrolytic enzymes, their characteristics, the possibilities of production and their applications. In the experimental part the production of hydrolytic enzymes by the fungus Penicillium lilacinum was performed in mineral medium and in a medium wherein as substrate yeast strains Sporobolomyces roseus and Rhodotorula glutinis were used. During cultivation process, production of cellulases, amylases, xylanases, lipases, proteases, and mannases was monitored. Samples were taken on 3rd, 5th, 8th, 10th and 12th day of cultivation. Production varies depending on time and substrate type. Amylase, cellulose and xylanase activity was measured in two steps - at the beginning of culture (3 to 5 day) and at the end of cultivation. The rest of enzymes were detected mainly in the 8th to 10th day of culture. The best three samples with the highest value of enzyme activities were further used for lyofilization and purification by ultrafiltration. Further, enzymes were tested for disruption of yeast cell wall. In the conclusion, the yeast protoplasts of yeast strains Rhodotorula glutinis a Sporobolomyces roseus were prepared successfully.
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Control of metabolism of carotenogenic yeasts on molecular level
Pokrývková, Zuzana ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
This diploma thesis deals with the molecular characterization of carotenogenic yeasts. The techniques used for the analysis of the conserved regions of the D1/D2 rDNA region of the 26S ribosomal large subunit region and the ITS1 and 5,8-ITS2 regions were nested PCR and DGGE. The results of DGGE show that all analyzed yeast strains have very similar sequences of these regions The yeast Rhodotorula mucilaginosa with the collection number CCY 20-7-28 showed differences from the other carotenogenic yeast strains. As a part of melucular characterisation using ribosomal gene sequences, eight yeast strains were examinated for substrate utilisation tests using different substrates. Characterisation of growth and metabolite production was tested in each strain too. The next aim of this thesis was to prepare a carotenoid yeast strain characterized by overproduction of metabolites, in particular carotenoids and lipids,. Yeasts were subjected to a random mutation caused by UV irradiation and the influence of this mutantagen onthe production of metabolites was evaluated. As a candidate yeast strain C. capitatum CCY 10-1-2 was selected. This selection was based on previous studies due to its good production of lipids using waste glycerol as asubstrate. This strain was subsequently adapted to waste whey, glycerol, and a glucose as a basic carbon source.
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Control of metabolism of carotenogenic yeasts on molecular level
Pokrývková, Zuzana ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
This diploma thesis deals with the molecular characterization of carotenogenic yeasts. The techniques used for the analysis of the conserved regions of the D1/D2 rDNA region of the 26S ribosomal large subunit region and the ITS1 and 5,8-ITS2 regions were nested PCR and DGGE. The results of DGGE show that all analyzed yeast strains have very similar sequences of these regions The yeast Rhodotorula mucilaginosa with the collection number CCY 20-7-28 showed differences from the other carotenogenic yeast strains. As a part of melucular characterisation using ribosomal gene sequences, eight yeast strains were examinated for substrate utilisation tests using different substrates. Characterisation of growth and metabolite production was tested in each strain too. The next aim of this thesis was to prepare a carotenoid yeast strain characterized by overproduction of metabolites, in particular carotenoids and lipids,. Yeasts were subjected to a random mutation caused by UV irradiation and the influence of this mutantagen onthe production of metabolites was evaluated. As a candidate yeast strain C. capitatum CCY 10-1-2 was selected. This selection was based on previous studies due to its good production of lipids using waste glycerol as asubstrate. This strain was subsequently adapted to waste whey, glycerol, and a glucose as a basic carbon source.
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Production of extracellular hydrolases by specific mould species
Pokrývková, Zuzana ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
This bachelor's thesis is focused on studying the possibilities of producing extracellular hydrolytic enzymes by the fungus Penicillium lilacinum on two substrates. The theoretical part is focused on the characterization of selected hydrolytic enzymes, their characteristics, the possibilities of production and their applications. In the experimental part the production of hydrolytic enzymes by the fungus Penicillium lilacinum was performed in mineral medium and in a medium wherein as substrate yeast strains Sporobolomyces roseus and Rhodotorula glutinis were used. During cultivation process, production of cellulases, amylases, xylanases, lipases, proteases, and mannases was monitored. Samples were taken on 3rd, 5th, 8th, 10th and 12th day of cultivation. Production varies depending on time and substrate type. Amylase, cellulose and xylanase activity was measured in two steps - at the beginning of culture (3 to 5 day) and at the end of cultivation. The rest of enzymes were detected mainly in the 8th to 10th day of culture. The best three samples with the highest value of enzyme activities were further used for lyofilization and purification by ultrafiltration. Further, enzymes were tested for disruption of yeast cell wall. In the conclusion, the yeast protoplasts of yeast strains Rhodotorula glutinis a Sporobolomyces roseus were prepared successfully.
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