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undergraduate thesis
Konstrukcija i mjerenje aktivnosti rekombinantnog proteina Pir2bla_CJ izloženog na površini stanične stijenke
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. urn:nbn:hr:159:194395
Matijević, Dominik
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

Matijević, D. (2022). Konstrukcija i mjerenje aktivnosti rekombinantnog proteina Pir2bla_CJ izloženog na površini stanične stijenke (Undergraduate thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:194395

Matijević, Dominik. "Konstrukcija i mjerenje aktivnosti rekombinantnog proteina Pir2bla_CJ izloženog na površini stanične stijenke." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. https://urn.nsk.hr/urn:nbn:hr:159:194395

Matijević, Dominik. "Konstrukcija i mjerenje aktivnosti rekombinantnog proteina Pir2bla_CJ izloženog na površini stanične stijenke." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. https://urn.nsk.hr/urn:nbn:hr:159:194395

Matijević, D. (2022). 'Konstrukcija i mjerenje aktivnosti rekombinantnog proteina Pir2bla_CJ izloženog na površini stanične stijenke', Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 29 June 2024, https://urn.nsk.hr/urn:nbn:hr:159:194395

Matijević D. Konstrukcija i mjerenje aktivnosti rekombinantnog proteina Pir2bla_CJ izloženog na površini stanične stijenke [Undergraduate thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2022 [cited 2024 June 29] Available at: https://urn.nsk.hr/urn:nbn:hr:159:194395

D. Matijević, "Konstrukcija i mjerenje aktivnosti rekombinantnog proteina Pir2bla_CJ izloženog na površini stanične stijenke", Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2022. Available at: https://urn.nsk.hr/urn:nbn:hr:159:194395

Metadata
TitleKonstrukcija i mjerenje aktivnosti rekombinantnog proteina Pir2bla_CJ izloženog na površini stanične stijenke
Title (english)Construction and activity determination of recombinant protein Pir2bla_CJ displayed on the yeast cell surface
AuthorDominik Matijević
MentorBojan Žunar (mentor)
GranterUniversity of Zagreb
Faculty of Food Technology and Biotechnology
(Department of Chemistry and Biochemistry)
(Laboratory for Biochemistry)
Zagreb
Defense date and country2022-06-29, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Jedna od alternativa klasičnim, unutarstaničnim, bioprocesima su izvanstanični bioprocesi temeljeni na sojevima kvasca koji izlažu biotehnološki zanimljive enzime na svojoj staničnoj površini. Takvo vezivanje proteina može se, na primjer, postići fuzijom enzima od interesa s proteinom Pir2. Međutim, nije poznato gdje je unutar proteina Pir2 najbolje umetnuti enzim, a da se što bolje sačuva i sposobnost vezanja proteina Pir2 i aktivnost izloženog enzima. U ovom radu konstruiran je plazmid pRSII-pir2bla_CJ koji kodira za fuzijski protein Pir2bla_CJ, spoj proteina Pir2 i enzima β-laktamaze. Nadalje, izmjerena je aktivnost β-laktamaze izložene na staničnoj stijenci kvasca Saccharomyces cerevisiae koji eksprimira ovaj rekombinantni protein i uspoređena s aktivnosti četiri prethodno konstruirane inačice proteina Pir2bla. Iako protein Pir2bla_CJ kombinira odlike dvije prethodno razvijene vrlo efikasne inačice proteina Pir2bla, njegova ekspresija rezultira zanemarivom aktivnošću β-laktamaze, sugerirajući da pravila koja diktiraju efikasno vezanje proteina na staničnu stijenku, uz očuvanje biotehnološki zanimljive enzimske aktivnosti, nisu aditivna.
Abstract (english)
Some classical, intracellular bioprocesses could be replaced with extracellular ones, i.e. bioprocesses based on yeast strains displaying biotechnologically interesting enzymes on their cell surface. We can bind such enzymes to the cell wall, for example, by fusing them with Pir2 proteins. However, it remains unclear where in the Pir2 protein we should insert the enzyme to preserve both the enzymatic activity and the Pir2 binding ability. In this work, we constructed a plasmid encoding protein Pir2bla_CJ, a fusion of the protein Pir2 and the enzyme β-lactamase. Furthermore, we measured the β-lactamase activity of Pir2bla_CJ on the Saccharomyces cerevisiae cell surface and compared it to four previously constructed variations of Pir2bla protein. Although Pir2bla_CJ combined characteristics of two high-performing Pir2bla variations, its β-lactamase activity was negligible, suggesting that the rules allowing for the robust binding of proteins on the cell wall, and preservation of biotechnologically relevant enzymatic activity, are not additive.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:159:194395
Study programmeTitle: Biotechnology
Study programme type: university
Study level: undergraduate
Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0) icon
Created on2022-10-05 12:35:25