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Pregled bibliografske jedinice broj: 1220940

Advancing Pir-based yeast surface display by reshuffling and truncating Pir2-β-lactamase constructs


Žunar, Bojan; Lozančić, Mateja; Matičević, Ana; Matijević, Dominik; Martinić Cezar, Tea; Teparić, Renata; Mrša, Vladimir
Advancing Pir-based yeast surface display by reshuffling and truncating Pir2-β-lactamase constructs // British Yeast Group 2022: From Cells to Genomes: Poster Book
London, Ujedinjeno Kraljevstvo, 2022. str. 3-3 (poster, recenziran, sažetak, znanstveni)


CROSBI ID: 1220940 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Advancing Pir-based yeast surface display by reshuffling and truncating Pir2-β-lactamase constructs

Autori
Žunar, Bojan ; Lozančić, Mateja ; Matičević, Ana ; Matijević, Dominik ; Martinić Cezar, Tea ; Teparić, Renata ; Mrša, Vladimir

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
British Yeast Group 2022: From Cells to Genomes: Poster Book / - , 2022, 3-3

Skup
British Yeast Group Meeting 2022: From Cells to Genomes

Mjesto i datum
London, Ujedinjeno Kraljevstvo, 07.09.2022. - 09.09.2022

Vrsta sudjelovanja
Poster

Vrsta recenzije
Recenziran

Ključne riječi
Pir proteins, cell wall, yeast, surface display, Saccharomyces cerevisiae

Sažetak
After synthesizing proteins, yeast Saccharomyces cerevisiae can retain them within the cell, secrete them into the medium, or bind them to the cell surface. In the third case, yeast anchors proteins by covalently binding them to β-glucan residues, thus allowing for surface display, a cell-engineering technique that can remake the entire cell surface into catalytically active living material. To immobilize proteins of interest closer to their N-terminus, the proteins are regularly fused with one of five Pir proteins (proteins with internal repeats), which covalently bind to β-1, 3-glucan. However, beyond the requirement for glutamine in their characteristic internal repeats, the mechanism of attaching these proteins to the yeast cell wall remains unclear. Thus, the Pir-based surface display continues to rely on guesswork and intuition, which is less than efficient. To address this issue, we inserted β-lactamase at five positions in Pir2 (Hsp150) protein and followed its activity and Pir2-binding efficiency through enzymatic and immunochemical methods. Moreover, we constructed and tested additional six truncated Pir2-β-lactamase variants, thus determining the minimal portion of Pir2 required for efficient binding to the yeast cell wall. Finally, to enrich our experimental results with structural insights, we used a deep- learning Alphafold2-based algorithm to in silico predict the structure of Pir2-β-lactamase fusions. Therefore, we present and rationalize a novel set of practical guidelines for a reproducible, straightforward, and efficient Pir-based yeast surface display.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija



POVEZANOST RADA


Projekti:
IP-2019-04-2891 - Biotehnološka primjena ugradnje heterolognih proteina u stanične stijenke kvasaca (PRODIS) (Mrša, Vladimir, HRZZ - 2019-04) ( CroRIS)

Ustanove:
Prehrambeno-biotehnološki fakultet, Zagreb


Citiraj ovu publikaciju:

Žunar, Bojan; Lozančić, Mateja; Matičević, Ana; Matijević, Dominik; Martinić Cezar, Tea; Teparić, Renata; Mrša, Vladimir
Advancing Pir-based yeast surface display by reshuffling and truncating Pir2-β-lactamase constructs // British Yeast Group 2022: From Cells to Genomes: Poster Book
London, Ujedinjeno Kraljevstvo, 2022. str. 3-3 (poster, recenziran, sažetak, znanstveni)
Žunar, B., Lozančić, M., Matičević, A., Matijević, D., Martinić Cezar, T., Teparić, R. & Mrša, V. (2022) Advancing Pir-based yeast surface display by reshuffling and truncating Pir2-β-lactamase constructs. U: British Yeast Group 2022: From Cells to Genomes: Poster Book.
@article{article, author = {\v{Z}unar, Bojan and Lozan\v{c}i\'{c}, Mateja and Mati\v{c}evi\'{c}, Ana and Matijevi\'{c}, Dominik and Martini\'{c} Cezar, Tea and Tepari\'{c}, Renata and Mr\v{s}a, Vladimir}, year = {2022}, pages = {3-3}, keywords = {Pir proteins, cell wall, yeast, surface display, Saccharomyces cerevisiae}, title = {Advancing Pir-based yeast surface display by reshuffling and truncating Pir2-β-lactamase constructs}, keyword = {Pir proteins, cell wall, yeast, surface display, Saccharomyces cerevisiae}, publisherplace = {London, Ujedinjeno Kraljevstvo} }
@article{article, author = {\v{Z}unar, Bojan and Lozan\v{c}i\'{c}, Mateja and Mati\v{c}evi\'{c}, Ana and Matijevi\'{c}, Dominik and Martini\'{c} Cezar, Tea and Tepari\'{c}, Renata and Mr\v{s}a, Vladimir}, year = {2022}, pages = {3-3}, keywords = {Pir proteins, cell wall, yeast, surface display, Saccharomyces cerevisiae}, title = {Advancing Pir-based yeast surface display by reshuffling and truncating Pir2-β-lactamase constructs}, keyword = {Pir proteins, cell wall, yeast, surface display, Saccharomyces cerevisiae}, publisherplace = {London, Ujedinjeno Kraljevstvo} }




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