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master's thesis
Pročišćavanje i karakterizacija novog fungalog ekstracelularnog enzima
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2016. urn:nbn:hr:159:929199
Lešić, Tanja
University of Zagreb
Faculty of Food Technology and Biotechnology
Department of Biochemical Engineering
Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology

Institutional repository: Repository of the Faculty of Food Technology and Biotechnology

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Lešić, T. (2016). Pročišćavanje i karakterizacija novog fungalog ekstracelularnog enzima (Master's thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:929199

Lešić, Tanja. "Pročišćavanje i karakterizacija novog fungalog ekstracelularnog enzima." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2016. https://urn.nsk.hr/urn:nbn:hr:159:929199

Lešić, Tanja. "Pročišćavanje i karakterizacija novog fungalog ekstracelularnog enzima." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2016. https://urn.nsk.hr/urn:nbn:hr:159:929199

Lešić, T. (2016). 'Pročišćavanje i karakterizacija novog fungalog ekstracelularnog enzima', Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 18 January 2025, https://urn.nsk.hr/urn:nbn:hr:159:929199

Lešić T. Pročišćavanje i karakterizacija novog fungalog ekstracelularnog enzima [Master's thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2016 [cited 2025 January 18] Available at: https://urn.nsk.hr/urn:nbn:hr:159:929199

T. Lešić, "Pročišćavanje i karakterizacija novog fungalog ekstracelularnog enzima", Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2016. Available at: https://urn.nsk.hr/urn:nbn:hr:159:929199

Metadata
TitlePročišćavanje i karakterizacija novog fungalog ekstracelularnog enzima
Title (english)Purification and characterization of a new fungal extracellular enzyme
AuthorTanja Lešić
MentorBožidar Šantek (mentor)
GranterUniversity of Zagreb
Faculty of Food Technology and Biotechnology
(Department of Biochemical Engineering)
(Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology)
Zagreb
Defense date and country2016-12-01, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Enzim celobioza dehidrogenaza iz Corynascus thermophilus (CtCDH) je eksprimiran u Aspergillus niger. Najbolji transformant je odabran za provjeru proizvodnje enzima, te za proizvodnju u Erlenmajerovim tikvicama. Ispitivanja tijekom proizvodnje pokazala su sličnost s divljim tipom CtCDH dok je CtCDH eksprimiranim u kvascu Pichia pastoris pokazao nižu specifičnu aktivnost. Enzim je pročišćen u dva stupnja. Molekularna masa iznosi 90 kDa. Kinetičke konstante enzima CtCDH su više u usporedbi s vrijednostima objavljenim za divlji tip CtCDH kao i za CtCDH exprimiran u Pichia pastoris. Enzimu CDH je također testirana sposobnost in situ proizvodnje H2O2 u reakciji s enzimom nespecifična peroksigenaza kako bi se uspostavio ekološki prihvatljiv proces oksidacije etilbenzena. U tu svrhu su testirani različiti CDH varijanti iz Myriococcum thermophilum sa povećanim afinitetom prema kisiku. Varijant N748G/ T750Q se pokazao kao najučinkovitiji. Ograničavajući faktor u primjeni ovakve vrste je stabilnost enzima, na što se buduća istraživanja moraju orijentirati.
Abstract (english)
Cellobiose dehydrogenase from Corynascus thermophilus (CtCDH) was cloned into Aspergillus niger for heterologous expression. The best transformant selected for screening was also selected for the enzyme production in Erlenmeyer flasks. During the production, the recombinant CtCDH showed similar characteristics to the wild-type CtCDH, whereas CtCDH produced in Pichia pastoris shows a lower specific activity. The expressed enzyme was purified in two-step procedure. The molecular mass was ~90 kDa. Kinetic constants of CtCDH were higher compared to the wild-type CtCDH and the one expressed in Pichia pastoris. CDH was also tested for its ability of the in situ production of H2O2 and it was combined with unspecific peroxygenase to establish an eco-friendly process for ethylbenzene oxidation. For that purpose, different CDH variants from Myriococcum thermophilum with increased oxygen reactivity have been tested. The variant N748G/ T750Q was found to be most efficient. A limiting factor in application of this type seems to be the enzyme's stability on which further research must focus.
Keywords
Keywords (english)
Languageenglish
URN:NBNurn:nbn:hr:159:929199
Study programmeTitle: Bioprocess Engineering
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva (magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva)
Type of resourceText
File originBorn digital
Access conditionsOpen access
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Created on2017-04-24 10:23:51