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undergraduate thesis
Ispitivanje utjecaja prekomjerne ekspresije kompleksa GPI-transamidaze na ugradnju Ccw12bla proteina u staničnu stijenku kvasca S. cerevisiae
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2024. urn:nbn:hr:159:573579
Pospiš, Hana
University of Zagreb
Faculty of Food Technology and Biotechnology
Department of Chemistry and Biochemistry
Laboratory for Biochemistry

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

Cite this document

Pospiš, H. (2024). Ispitivanje utjecaja prekomjerne ekspresije kompleksa GPI-transamidaze na ugradnju Ccw12bla proteina u staničnu stijenku kvasca S. cerevisiae (Undergraduate thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:573579

Pospiš, Hana. "Ispitivanje utjecaja prekomjerne ekspresije kompleksa GPI-transamidaze na ugradnju Ccw12bla proteina u staničnu stijenku kvasca S. cerevisiae." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2024. https://urn.nsk.hr/urn:nbn:hr:159:573579

Pospiš, Hana. "Ispitivanje utjecaja prekomjerne ekspresije kompleksa GPI-transamidaze na ugradnju Ccw12bla proteina u staničnu stijenku kvasca S. cerevisiae." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2024. https://urn.nsk.hr/urn:nbn:hr:159:573579

Pospiš, H. (2024). 'Ispitivanje utjecaja prekomjerne ekspresije kompleksa GPI-transamidaze na ugradnju Ccw12bla proteina u staničnu stijenku kvasca S. cerevisiae', Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 30 November 2024, https://urn.nsk.hr/urn:nbn:hr:159:573579

Pospiš H. Ispitivanje utjecaja prekomjerne ekspresije kompleksa GPI-transamidaze na ugradnju Ccw12bla proteina u staničnu stijenku kvasca S. cerevisiae [Undergraduate thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2024 [cited 2024 November 30] Available at: https://urn.nsk.hr/urn:nbn:hr:159:573579

H. Pospiš, "Ispitivanje utjecaja prekomjerne ekspresije kompleksa GPI-transamidaze na ugradnju Ccw12bla proteina u staničnu stijenku kvasca S. cerevisiae", Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2024. Available at: https://urn.nsk.hr/urn:nbn:hr:159:573579

Metadata
TitleIspitivanje utjecaja prekomjerne ekspresije kompleksa GPI-transamidaze na ugradnju Ccw12bla proteina u staničnu stijenku kvasca S. cerevisiae
Title (english)Examination of the effect of overexpression of the GPI-transamidase complex on the incorporation of Ccw12bla protein into the cell wall of the yeast S. cerevisiae
AuthorHana Pospiš
MentorRenata Teparić (mentor)
MentorTea Martinić Cezar (pomoć pri izradi, neposredni voditelj)
GranterUniversity of Zagreb
Faculty of Food Technology and Biotechnology
(Department of Chemistry and Biochemistry)
(Laboratory for Biochemistry)
Zagreb
Defense date and country2024-09-16, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Food Technology
Abstract
„Yeast surface display“ (YSD) sustav jednostavnija je, efikasnija i jeftinija metoda proizvodnje i imobilizacije enzima, nego klasične imobilizacije u kojima se za vezanje proteina na neki netopljivi nosač koriste kemikalije i postupci koje često imaju negativan utjecaj na enzimsku aktivnost. Rekombinantni protein mora sadržati protein od interesa fuzioniran sa nativnim proteinom stanične stijenke. Nedostatci ove metode su potencijalno smanjenje aktivnosti proteina i neuspješna ugradnja na površinu stanice. Bitna prepreka za industrijsku primjenu ove metode je njena još uvijek niska efikasnosti. Stoga se i dalje radi na tome da se efikasnost YSD poveća. Cilj ovog rada jest ispitati utjecaj prekomjerne ekspresije kompleksa GPI-transamidaze na ugradnju Ccw12bla proteina na površinu stanice. Ccw12bla protein eksprimiran je u stanicama kvasca divljeg tipa i u stanicama koje su bile transformirane plazmidima koji su sadržali genetičku informaciju za povećanu ekspresiju podjedinica kompleksa GPI-transamidaze. Uspješnost provedenog eksperimenta ispitana je mjerenjem aktivnosti β-laktamaze i imunoblotom ukupnih proteina stijenke.
Abstract (english)
“Yeast surface display" (YSD) system is a simpler, more efficient and cheaper method of enzyme production and immobilization than classic immobilizations in which chemicals and procedures used to attach proteins to some insoluble carrier often have a negative effect on enzyme activity. The recombinant protein must contain protein of interest fused to some native cell wall protein. The disadvantages of this method are the potential reduction of protein activity and unsuccessful incorporation on the cell surface. Low efficiency of YSD remains a significant obstacle for industrial application of this method, and efforts are still being made to improve its efficiency. The aim of this work is to examine the influence of overexpression of the GPI-transamidase complex on the incorporation of the Ccw12bla protein on the cell surface. The Ccw12bla protein was expressed in wild-type yeast cells and in cells transformed with plasmids containing genetic information for overexpression of the GPI transamidase complex subunits. The success of the conducted experiment was tested by measuring β-lactamase activity and immunoblot of cell wall proteins.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:159:573579
Study programmeTitle: Food Technology
Study programme type: university
Study level: undergraduate
Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka prehrambene tehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka prehrambene tehnologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0) icon
Created on2024-09-25 11:03:03