prikaz prve stranice dokumenta Production of glucose dehydrogenase/cytochrome chimeras by sortagging and characterization of FAD binding constants in GMC-oxidoreductases
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master's thesis
Production of glucose dehydrogenase/cytochrome chimeras by sortagging and characterization of FAD binding constants in GMC-oxidoreductases
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2021. urn:nbn:hr:159:044091
Nikolić, Lorna
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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Nikolić, L. (2021). Production of glucose dehydrogenase/cytochrome chimeras by sortagging and characterization of FAD binding constants in GMC-oxidoreductases (Master's thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:044091

Nikolić, Lorna. "Production of glucose dehydrogenase/cytochrome chimeras by sortagging and characterization of FAD binding constants in GMC-oxidoreductases." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2021. https://urn.nsk.hr/urn:nbn:hr:159:044091

Nikolić, Lorna. "Production of glucose dehydrogenase/cytochrome chimeras by sortagging and characterization of FAD binding constants in GMC-oxidoreductases." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2021. https://urn.nsk.hr/urn:nbn:hr:159:044091

Nikolić, L. (2021). 'Production of glucose dehydrogenase/cytochrome chimeras by sortagging and characterization of FAD binding constants in GMC-oxidoreductases', Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 06 June 2024, https://urn.nsk.hr/urn:nbn:hr:159:044091

Nikolić L. Production of glucose dehydrogenase/cytochrome chimeras by sortagging and characterization of FAD binding constants in GMC-oxidoreductases [Master's thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2021 [cited 2024 June 06] Available at: https://urn.nsk.hr/urn:nbn:hr:159:044091

L. Nikolić, "Production of glucose dehydrogenase/cytochrome chimeras by sortagging and characterization of FAD binding constants in GMC-oxidoreductases", Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:159:044091

Metadata
TitleProduction of glucose dehydrogenase/cytochrome chimeras by sortagging and characterization of FAD binding constants in GMC-oxidoreductases
Title (croatian)Proizvodnja glukoza dehidrogenaza/citokrom kimernog enzima pomoću sortaze i karakterizacija konstanti vezanja FAD-a kod GMC-oksidoreduktaza
AuthorLorna Nikolić
MentorTonči Rezić (mentor)
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 country2021-09-29, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Majority of the growing market for biosensors is driven by medical diagnostics, especially glucose biosensors. Currently, the development of third generation mediatorless biosensors for glucose monitoring is underway. These biosensors employ GMC oxidoreductases as a biological recognition element and an electrochemical transducer for mediatorless detection and quantification of glucose. The problems of direct electron transfer (DET) to electrode, as well as low stability and substrate specificity still have to be overcome and were assesed in this thesis. In order to achieve DET, glucose dehydrogenase (GDH) and cytochrome c were postranslationally conjugated by sortase A to create a chimeric enzyme of improved perfomance mimicking DET capable cellobiose dehydrogenase (CDH). Furthermore, FAD binding constants of native AnGOx, as well as recombinant AnGOX, GcGDH and MtDH, expressed in P. pastoris X33, were determined by deflavination, followed by reflavination of apoenzymes with synthetic FAD. The goal was to create fusion protein capable of DET and to gain insights into the coffactor affinity
Abstract (croatian)
Većinski udio u rastućem tržištu biosenzora zauzimaju potrebe medicinske dijagnostike za detekciju razine glukoze u krvi. Danas se aktivno radi na razvoju glukoznog biosenzora treće generacije, sastavni dio kojeg su GMC oksidoreduktaze, koje provode biokemijsku reakciju i pomoću elektrokemijskog transduktora (pretvarača signala), direktno i bez nusprodukata, prenose elektrone na elektrodu i omogućavaju detekciju i kvantifikaciju glukoze. Problemi koje je potrebno riješiti da bi ova tehnologija bila što pouzdanija su direktni elektronski transfer (DET) s enzima na elektrodu, te niska stabilnost i specifičnost enzima za supstrat. U svrhu postizanja DET-a proizveden je kimerni enzim koji oponaša elektronski transfer celobioza dehidrogenaze (CDH) povezivanjem glukoza dehidrogenaze (GDH) i citokroma c posttranslacijskom konjugacijom posredstvom sortaze A. Također, ispitane su i uspoređene konstante vezanja kofatora FAD-a GMC oksidoreduktaza od interesa (nativni AnGOX, i rekombinantni AnGOX, GcGDH, MtDH, proizvedeni pomoću P. pastoris X33) deflavinacijom i reflavinacijom pomoću sintetskog FAD-a. Cilj je bio proizvodnja sintetskog fuzijskog proteina s mogućnošću DET-a kako bi stekli uvid u afinitet kofaktora.
Keywords
Keywords (english)
Languageenglish
URN:NBNurn:nbn:hr:159:044091
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
Terms of useLicence Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0) icon
Created on2022-05-26 11:16:56