Produkce extracelulárních hydroláz pomocí specifických druhů plísní
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Vysoké učení technické v Brně. Fakulta chemická
Abstract
Táto bakalárska práca je zameraná na štúdium možností produkcie extracelulárnych hydrolytických enzýmov plesňou Penicillium lilacinum na dvoch substrátoch. Teoretická časť práce je zameraná na charakterizáciu vybraných hydrolytických enzýmov, ich vlastnosti, možnosť produkcie a ich aplikácie. V experimentálnej časti boli hydrolytické enzýmy produkované plesňou Penicillium lilacinum pri statickej kultivácií v minerálnom médiu a v médiu kde ako substrát boli použité kmene kvasiniek Sporobolomyces roseus a Rhodotorula glutinis. V priebehu kultivácie bola sledovaná produkcia celuláz, amyláz, xylanáz, lipáz, proteáz a manáz a to vždy 3., 5., 8., 10. a 12. deň kultivácie. Produkcia sa líši v závislosti na čase a použitom substráte. Amylázová, celulázová, xylanázová aktivita bola meraná v dvoch periódach - na začiatku kultivácie (3. až 5. deň) a na konci kultivácie. Zvyšné enzýmy boli detegované najmä v 8. až 10. deň kultivácie. Tri najlepšie vzorky s najvyššou hodnotou enzýmových aktivít boli ďalej použité k testovaniu lyofilizácie a k purifikácií pomocou ultrafiltrácie. Enzýmy ďalej boli testované k rozbíjaniu bunkovej steny kvasiniek. V závere práce boli úspešne pripravené protoplasty kvasiniek Rhodotorula glutinis a Sporobolomyces roseus.
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.
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.
Description
Citation
POKRÝVKOVÁ, Z. Produkce extracelulárních hydroláz pomocí specifických druhů plísní [online]. Brno: Vysoké učení technické v Brně. Fakulta chemická. 2015.
Document type
Document version
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Language of document
cs
Study field
Biotechnologie
Comittee
prof. RNDr. Ivana Márová, CSc. (předseda)
prof. Ing. Peter Šimko, DrSc. (místopředseda)
doc. Ing. Jiřina Omelková, CSc. (člen)
doc. RNDr. Alena Španová, CSc. (člen)
doc. Ing. Bohuslav Rittich, CSc. (člen)
doc. Ing. Pavel Diviš, Ph.D. (člen)
Date of acceptance
2015-06-17
Defence
1. Studentka seznámila členy komise s náplní a cílem bakalářské práce.
2. Byly přečteny posudky na bakalářskou práci.
3. Studentka akceptovala všechny připomínky oponenta a na všechny otázky odpověděla v plné šíři.
Diskuse
doc. Španová: Příprava protoplastů ?
Result of defence
práce byla úspěšně obhájena
Document licence
Standardní licenční smlouva - přístup k plnému textu bez omezení