prikaz prve stranice dokumenta Identification and characterization of cellodextrin phosphorylases for their application in synthesis of reducing-end thiol-modified cello-oligosaccharides
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master's thesis
Identification and characterization of cellodextrin phosphorylases for their application in synthesis of reducing-end thiol-modified cello-oligosaccharides
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2023. urn:nbn:hr:159:241474
Sušac, Mislav
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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Sušac, M. (2023). Identification and characterization of cellodextrin phosphorylases for their application in synthesis of reducing-end thiol-modified cello-oligosaccharides (Master's thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:241474

Sušac, Mislav. "Identification and characterization of cellodextrin phosphorylases for their application in synthesis of reducing-end thiol-modified cello-oligosaccharides." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2023. https://urn.nsk.hr/urn:nbn:hr:159:241474

Sušac, Mislav. "Identification and characterization of cellodextrin phosphorylases for their application in synthesis of reducing-end thiol-modified cello-oligosaccharides." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2023. https://urn.nsk.hr/urn:nbn:hr:159:241474

Sušac, M. (2023). 'Identification and characterization of cellodextrin phosphorylases for their application in synthesis of reducing-end thiol-modified cello-oligosaccharides', Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 30 August 2024, https://urn.nsk.hr/urn:nbn:hr:159:241474

Sušac M. Identification and characterization of cellodextrin phosphorylases for their application in synthesis of reducing-end thiol-modified cello-oligosaccharides [Master's thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2023 [cited 2024 August 30] Available at: https://urn.nsk.hr/urn:nbn:hr:159:241474

M. Sušac, "Identification and characterization of cellodextrin phosphorylases for their application in synthesis of reducing-end thiol-modified cello-oligosaccharides", Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2023. Available at: https://urn.nsk.hr/urn:nbn:hr:159:241474

Metadata
TitleIdentification and characterization of cellodextrin phosphorylases for their application in synthesis of reducing-end thiol-modified cello-oligosaccharides
Title (croatian)Identifikacija i karakterizacija celodekstrin fosforilaza za njihovu primjenu u sintezi tiol-modificiranih celo-oligosaharida
AuthorMislav Sušac
MentorAnita Slavica (mentor)
Committee memberRenata Teparic (predsjednik povjerenstva)
Committee memberBlaženka Kos (član povjerenstva)
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 country2023-03-15, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Four promising cellodextrin phosphorylases (CdPs) were identified as potential candidates for synthesis of reducing-end thiol-modified cello-oligosaccharides (retmco). The enzymes catalyzed two reactions - synthesis of the retmco and hydrolysis of glucosyl donor. CdP from Spirochetae thermophila (StCdP) proved to be the best candidate (the highest synthesis and the lowest hydrolase activity). Further improvement was achieved by the dual-enzyme cascade reaction, performed by cellobiose phosphorylase from Clostridium udae (CuCbP) and StCdP. The purity of synthesized 1-thio-cello-oligosaccharides >95% was confirmed with 1H-NMR, and average degree of polymerization was estimated to be 9.3. CdPs with low hydrolase activity have tyrosine in catalytic loop adjacent to the active site, while CdPs with high hydrolase activity have cysteine in this specific position, as confirmed by three single-site mutants. Additional improvement in the synthesis of the retmco was attained by the next dual-enzyme cascade reaction, performed by CuCbP and CdP from Clostridium cellulose mutant C626T.
Abstract (croatian)
Identificirane su četiri celodekstrin fosforilaze (CdPs) kao potencijalni kandidati za sintezu tiol-modificiranih celo-oligosaharida (retmco). Ovi enzimi kataliziraju reakciju u dva smjera - sintezu retmco i hidrolizu molekule donora glukozilne podjedinice. Ponajbolji kandidat (koji je imao najvišu aktivnost sintetaze i najnižu hidrolaznu aktivnost) je bila CdP iz Spirochetae thermophila (StCdP). Daljnje poboljšanje sinteze retmco postignuto je primjenom kaskadne reakcije s dva enzima - celobioza fosforilazom iz Clostridium udae (CuCbP) i StCdP. Čistoća sintetiziranog 1-tio-celo-oligosaharida iznad 95 % je potvrđena 1H-NMR analizom i prosječni stupanj polimerizacije dobivenog 1-tio-celooligosaharida procijenjen je na 9,3. CdPs sa niskom hidrolaznom aktivnosti imaju tirozin u katalitičkoj omči blizu aktivnog mjesta enzima, dok CdPs sa visokom hidrolaznom aktivnosti imaju cistein na ovoj specifičnoj poziciji, kao što je potvrđeno s tri točkaste mutacije. Daljnje poboljšanje sinteze retmco realizirano je primjenom kaskadne reakcije s dva enzima - CuCbP i mutanta C626T CdP iz Clostridium cellulose.
Keywords
Keywords (croatian)
Languageenglish
URN:NBNurn:nbn:hr:159:241474
Study programmeTitle: Molecular Biotechnology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
Type of resourceText
File originBorn digital
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Created on2024-06-27 10:16:58