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master's thesis
Production and characterization of different variants of Myriococcum thermophilum cellobiose dehydrogenase with the aim of improving cofactor binding and kinetic stability
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. urn:nbn:hr:159:847876
Ilić, Mario
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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Ilić, M. (2022). Production and characterization of different variants of Myriococcum thermophilum cellobiose dehydrogenase with the aim of improving cofactor binding and kinetic stability (Master's thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:847876

Ilić, Mario. "Production and characterization of different variants of Myriococcum thermophilum cellobiose dehydrogenase with the aim of improving cofactor binding and kinetic stability." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. https://urn.nsk.hr/urn:nbn:hr:159:847876

Ilić, Mario. "Production and characterization of different variants of Myriococcum thermophilum cellobiose dehydrogenase with the aim of improving cofactor binding and kinetic stability." Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. https://urn.nsk.hr/urn:nbn:hr:159:847876

Ilić, M. (2022). 'Production and characterization of different variants of Myriococcum thermophilum cellobiose dehydrogenase with the aim of improving cofactor binding and kinetic stability', Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 04 July 2024, https://urn.nsk.hr/urn:nbn:hr:159:847876

Ilić M. Production and characterization of different variants of Myriococcum thermophilum cellobiose dehydrogenase with the aim of improving cofactor binding and kinetic stability [Master's thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2022 [cited 2024 July 04] Available at: https://urn.nsk.hr/urn:nbn:hr:159:847876

M. Ilić, "Production and characterization of different variants of Myriococcum thermophilum cellobiose dehydrogenase with the aim of improving cofactor binding and kinetic stability", Master's thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2022. Available at: https://urn.nsk.hr/urn:nbn:hr:159:847876

Metadata
TitleProduction and characterization of different variants of Myriococcum thermophilum cellobiose dehydrogenase with the aim of improving cofactor binding and kinetic stability
Title (croatian)Proizvodnja i karakterizacija mutanata Myriococcum thermophilum celobioza dehidrogenaze s ciljem povećanja vezanja kofaktora i kinetičke stabilnosti
AuthorMario Ilić
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 country2022-02-04, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Biosensors are being extensively used in the healthcare sector but mostly in blood glucose monitoring for early diagnosis of diabetes. The use of enzymes as biological recognition elements is hampered in many ways, e.g., by insufficient communication between enzyme and electrode, low activity and stability. Cellobiose dehydrogenase (CDH) is a flavocytochrome enzyme that in native wild-type form shows efficient direct electron transfer (DET). To alter FAD cofactor binding and kinetic stability, mutations were introduced into the Myriococcum thermophilum CDH gene by site-directed mutagenesis. Variants were expressed in X-33 Pichia pastoris cells, purified and characterized. With an increased specific activity, catalytic efficiency, heme reduction rate and increased melting temperatures in pH range 5.0-8.0 the T306I variant seems to be the most promising variant for further investigation.
Abstract (croatian)
Biosenzori se uvelike koriste u zdravstvenom sektoru, a najveću primjenu pronalaze u praćenju i kontroli šećera glukoze oboljelih od dijabetesa. Uporaba enzima u biosenzorima je ograničena nedovoljno efikasnim direktnim prijenosom elektrona na elektrodu, niskom specifičnom aktivnosti i stabilnosti kroz duži vremenski period. Celobioza dehidrogenaza (CDH) je flavocitokrom koji u svojoj izvornoj formi ima mogućnost direktnog prijenosa elektrona (DET). S ciljem povećanja vezanja FAD koenzima i kinetičke aktivnosti, uvedene su mutacije u CDH gen izoliran iz askomicete Myriococcum thermophilum. Enzimi proizvedeni u soju kvasca Pichia pastoris X-33 su pročišćeni i karakterizirani. Mutant T306I pokazuje zadovoljavajuća svojstva (povećanu specifičnu aktivnost, kinetičku efikasnost, brzinu redukcije hema i temperaturu prijelaza) u području pH vrijednosti 5,0-8,0, stoga je potrebno provesti dodatna istraživanja korištenjem mutanta T306I.
Keywords
Keywords (english)
Languageenglish
URN:NBNurn:nbn:hr:159:847876
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 on2022-05-27 09:12:10