Pregled bibliografske jedinice broj: 1273951
Utilizing Ccw12 as an effective anchor for yeast surface display of diverse recombinant proteins
Utilizing Ccw12 as an effective anchor for yeast surface display of diverse recombinant proteins // Power of Microbesin Industry and Environment, BOOK OF ABSTRACTS / Teparić, Renata ; Leboš Pavunc, Andreja ; Kifer, Domagoj (ur.).
Zagreb: Hrvatsko mikrobiološko društvo, 2023. str. 113-113 (poster, nije recenziran, sažetak, znanstveni)
CROSBI ID: 1273951 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Utilizing Ccw12 as an effective anchor for yeast surface display of diverse recombinant proteins
Autori
Martinić Cezar, Tea ; Lozančić, Mateja ; Žunar, Bojan ; Mrša, Vladimir ; Teparić, Renata
Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni
Izvornik
Power of Microbesin Industry and Environment, BOOK OF ABSTRACTS
/ Teparić, Renata ; Leboš Pavunc, Andreja ; Kifer, Domagoj - Zagreb : Hrvatsko mikrobiološko društvo, 2023, 113-113
ISBN
978-953-7778-19-4
Skup
Power of Microbes in Industry and Environment 2023
Mjesto i datum
Poreč, Hrvatska, 15.05.2023. - 18.05.2023
Vrsta sudjelovanja
Poster
Vrsta recenzije
Nije recenziran
Ključne riječi
yeast surface display ; S.cerevisiae ; GPI anchor
Sažetak
Yeast surface display is an innovative technique that can improve the stability and activity of recombinant enzymes by attaching the enzymes to the yeast cell wall. In this study, we focused on the C-terminal immobilization of recombinant proteins, as certain proteins require exclusive immobilization through their C-terminus to prevent loss of activity. C-terminal immobilization via the anchor Ccw12 protein has been a promising approach for the efficient display of various industrially relevant recombinant proteins due to the high abundance and stable attachment of the Ccw12 protein in the yeast cell wall. To further enhance the efficiency of C-terminal immobilization, we conducted a study where we examined a series of strains in which genes associated with the regulation of transcription of genes related to wall structure maintenance, mRNA processing, translation, quality control, and maintenance of ER morphology were deleted. We investigated the activity of the reporter Ccw12BLA protein in these mutants and compared it to the wild-type strain. Increased Ccw12BLA activity was observed in mutants lacking protein involved in amino acid deprivation response (gcn2), translational repressor for cell wall proteins (ssd1), a protein involved in endoplasmic reticulum-associated protein degradation pathway (doa10), and covalently linked proteins of the cell wall (scw4pir1pir2pir3pir4). Additionally, Ccw12BLA optimization was applied to the surface display of sucrose phosphorylase, an enzyme that plays a role in glucosyl glycerol synthesis and has extensive applications in various industries including cosmetics, food, and pharmaceuticals. Sucrose phosphorylase (kindly provided by Prof. B. Nidetzky, TU Graz) was successfully expressed and immobilized through the Ccw12 anchor protein on the yeast cell wall in gcn2, doa10, ssd1, and scw4pir1pir2pir3pir4 mutants, and its binding was confirmed through Western blot analysis. The activity measurement of Ccw12SP in these mutants showed a similar trend as observed for the Ccw12BLA recombinant protein, suggesting that these mutants are promising candidates for glucosyl glycerol production using yeast surface display technology. Also, we expressed methionine adenosyltransferase, a key enzyme in S-adenosylmethionine production, and confirmed its localization on the yeast cell wall.
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
Profili:
Bojan Žunar
(autor)
Vladimir Mrša
(autor)
Tea Martinić-Cezar
(autor)
Renata Teparić
(autor)
Mateja Lozančić
(autor)