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undergraduate thesis
Utjecaj uklanjanja Cwp2, Scw4 i/ili Pir proteina iz stanične stijenke na efikasnost izlaganja heterolognih proteina na površini kvasca
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. urn:nbn:hr:159:845886
Vukšić, Lucija
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

Vukšić, L. (2022). Utjecaj uklanjanja Cwp2, Scw4 i/ili Pir proteina iz stanične stijenke na efikasnost izlaganja heterolognih proteina na površini kvasca (Undergraduate thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:845886

Vukšić, Lucija. "Utjecaj uklanjanja Cwp2, Scw4 i/ili Pir proteina iz stanične stijenke na efikasnost izlaganja heterolognih proteina na površini kvasca." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. https://urn.nsk.hr/urn:nbn:hr:159:845886

Vukšić, Lucija. "Utjecaj uklanjanja Cwp2, Scw4 i/ili Pir proteina iz stanične stijenke na efikasnost izlaganja heterolognih proteina na površini kvasca." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2022. https://urn.nsk.hr/urn:nbn:hr:159:845886

Vukšić, L. (2022). 'Utjecaj uklanjanja Cwp2, Scw4 i/ili Pir proteina iz stanične stijenke na efikasnost izlaganja heterolognih proteina na površini kvasca', Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 15 July 2024, https://urn.nsk.hr/urn:nbn:hr:159:845886

Vukšić L. Utjecaj uklanjanja Cwp2, Scw4 i/ili Pir proteina iz stanične stijenke na efikasnost izlaganja heterolognih proteina na površini kvasca [Undergraduate thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2022 [cited 2024 July 15] Available at: https://urn.nsk.hr/urn:nbn:hr:159:845886

L. Vukšić, "Utjecaj uklanjanja Cwp2, Scw4 i/ili Pir proteina iz stanične stijenke na efikasnost izlaganja heterolognih proteina na površini kvasca", Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2022. Available at: https://urn.nsk.hr/urn:nbn:hr:159:845886

Metadata
TitleUtjecaj uklanjanja Cwp2, Scw4 i/ili Pir proteina iz stanične stijenke na efikasnost izlaganja heterolognih proteina na površini kvasca
Title (english)The effect of removing Cwp2, Scw4 and/or Pir proteins from the cell wall on the efficiency of yeast surface display of heterologus proteins
AuthorLucija Vukšić
MentorMateja Lozančić (mentor)
GranterUniversity of Zagreb
Faculty of Food Technology and Biotechnology
(Department of Chemistry and Biochemistry)
(Laboratory for Biochemistry)
Zagreb
Defense date and country2022-09-07, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Tehnologija izlaganja proteina na površinu stanične stijenke temelji se na fuzioniranju gena od interesa s nekim od gena koji kodira za proteine stijenke te se tako nastali rekombinantni protein ugrađuje u stijenku kvasca. Na ovaj način moguće je eksprimirati čitave enzimske sustave čime se postiže njihova imobilizacija, lakše izdvajanje produkta te produljeno prisustvo biokatalizatora u odnosu na procese koji koriste pročišćene enzime. Cilj ovog rada jest ispitati utjecaj uklanjanja nativnih proteina iz stanične stijenke na efikasnost izlaganja heterolognih proteina na površinu stanice. U tu svrhu korišteni su konstrukti u kojima je gen bla fuzioniran sa nativnim genom PIR2 odnosno dijelom gena CCW12 te je rekombinantni protein zatim eksprimiran u stanicama divljeg tipa i u stanicama mutanata. Uspješnost ugradnje enzima u staničnu stijenku analizirana je mjerenjem aktivnosti βlaktamaze izložene na površinu stanice. Najveće povećanje aktivnosti u odnosu na stanice divljeg tipa postignuto je eksprimiranjem konstrukta CCW12bla u stanicama 4pirΔscw4Δ .
Abstract (english)
The yeast surface display technology is based on fusing the gene of interest with one of the genes that code for the proteins of the cell wall, and the resulting recombinant protein is incorporated in the wall. By using surface display it is posible to achieve expresion and imobilisation of enzymes, simplification of product purification, and prolongation of biocatalysts presence in the reactions. This work aims to examine the influence of removing native proteins from the cell wall on the efficiency of surface display. For this purpose, constructs were used in which the bla gene, coding for beta-lactamase, was fused with the PIR2 or fragment of CCW12 gene. The recombinant protein was expressed in wildtype and mutated cells. Efficiency of enzyme incorporation into the wall was analyzed by measuring βlactamase activity. Highest increase of activity compaired to wild type was measured in 4pirΔscw4Δ mutant when CCW12bla construct was expressed.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:159:845886
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 on2024-04-08 09:13:41