Glikozilacija i pH stabilnost penicilin G acilaze iz providencia rettgeri proizvedene u Pichia pastoris
Glycosylation and pH stability of penicillin G acylase from providencia rettgeri produced in Pichia pastoris
Апстракт
Penicilin G acilaza (PAC) je jedan od najšire korišćenih enzima u industrijskoj sintezi polusintetskih antibiotika. U ovom radu dobijeni nivo ekspresije PAC gena iz Providencia rettgeri u ekspresionom sistemu Pichia pastoris iznosio je 2.7 U/ml. Rekombinantni enzim je prečišćen i određen je njegov glikozilacioni status. Nađeno je da osim što su obe subjedinice enzima (α i β) N-glikozilovane, β subjedinica sadrži još i O-glikane. Takođe je ustanovljeno da je rekombinantna PACP. rett. stabilna u širokom pH opsegu što ju je, zajedno sa predhodno ustanovljenom visokom termostabilnošću, učinilo izuzetno privlačnim biokatalizatorom sa industrijske tačke gledišta.
Penicillin G acylase (PAC) is one of the most widely used enzymes in industrial synthesis of semi-synthetic antibiotics. The Providencia rettgeri pac gene was expressed to a level of 2.7 U/ml using the Pichia pastoris expression system. The recombinant enzyme was purified and its glycosylation status was determined. It was found that both subunits (α and β) of the enzyme were N-glycosylated, while the β-subunit also contained O-glycans. It was also observed that rPACP.rett. was stable in a wide range of pH, which, in addition to the previously proved high thermostability, makes it an attractive biocatalyst from an industrial point of view.
Кључне речи:
Pichia pastoris / pH stability / Penicillin acylase / glycosylationИзвор:
Archives of Biological Sciences, 2009, 61, 4, 581-586Издавач:
- Srpsko biološko društvo, Beograd, i dr.
Финансирање / пројекти:
- Експресија и регулација фармацеутски значајних гена у микроорганизмима (RS-MESTD-MPN2006-2010-143056)
DOI: 10.2298/ABS0904581S
ISSN: 0354-4664
WoS: 000273203500002
Scopus: 2-s2.0-77949907121
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Šenerović, Lidija AU - Stanković, Nada AU - Ljubijankić, G. AU - Vasiljević, Branka PY - 2009 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/381 AB - Penicilin G acilaza (PAC) je jedan od najšire korišćenih enzima u industrijskoj sintezi polusintetskih antibiotika. U ovom radu dobijeni nivo ekspresije PAC gena iz Providencia rettgeri u ekspresionom sistemu Pichia pastoris iznosio je 2.7 U/ml. Rekombinantni enzim je prečišćen i određen je njegov glikozilacioni status. Nađeno je da osim što su obe subjedinice enzima (α i β) N-glikozilovane, β subjedinica sadrži još i O-glikane. Takođe je ustanovljeno da je rekombinantna PACP. rett. stabilna u širokom pH opsegu što ju je, zajedno sa predhodno ustanovljenom visokom termostabilnošću, učinilo izuzetno privlačnim biokatalizatorom sa industrijske tačke gledišta. AB - Penicillin G acylase (PAC) is one of the most widely used enzymes in industrial synthesis of semi-synthetic antibiotics. The Providencia rettgeri pac gene was expressed to a level of 2.7 U/ml using the Pichia pastoris expression system. The recombinant enzyme was purified and its glycosylation status was determined. It was found that both subunits (α and β) of the enzyme were N-glycosylated, while the β-subunit also contained O-glycans. It was also observed that rPACP.rett. was stable in a wide range of pH, which, in addition to the previously proved high thermostability, makes it an attractive biocatalyst from an industrial point of view. PB - Srpsko biološko društvo, Beograd, i dr. T2 - Archives of Biological Sciences T1 - Glikozilacija i pH stabilnost penicilin G acilaze iz providencia rettgeri proizvedene u Pichia pastoris T1 - Glycosylation and pH stability of penicillin G acylase from providencia rettgeri produced in Pichia pastoris EP - 586 IS - 4 SP - 581 VL - 61 DO - 10.2298/ABS0904581S ER -
@article{ author = "Šenerović, Lidija and Stanković, Nada and Ljubijankić, G. and Vasiljević, Branka", year = "2009", abstract = "Penicilin G acilaza (PAC) je jedan od najšire korišćenih enzima u industrijskoj sintezi polusintetskih antibiotika. U ovom radu dobijeni nivo ekspresije PAC gena iz Providencia rettgeri u ekspresionom sistemu Pichia pastoris iznosio je 2.7 U/ml. Rekombinantni enzim je prečišćen i određen je njegov glikozilacioni status. Nađeno je da osim što su obe subjedinice enzima (α i β) N-glikozilovane, β subjedinica sadrži još i O-glikane. Takođe je ustanovljeno da je rekombinantna PACP. rett. stabilna u širokom pH opsegu što ju je, zajedno sa predhodno ustanovljenom visokom termostabilnošću, učinilo izuzetno privlačnim biokatalizatorom sa industrijske tačke gledišta., Penicillin G acylase (PAC) is one of the most widely used enzymes in industrial synthesis of semi-synthetic antibiotics. The Providencia rettgeri pac gene was expressed to a level of 2.7 U/ml using the Pichia pastoris expression system. The recombinant enzyme was purified and its glycosylation status was determined. It was found that both subunits (α and β) of the enzyme were N-glycosylated, while the β-subunit also contained O-glycans. It was also observed that rPACP.rett. was stable in a wide range of pH, which, in addition to the previously proved high thermostability, makes it an attractive biocatalyst from an industrial point of view.", publisher = "Srpsko biološko društvo, Beograd, i dr.", journal = "Archives of Biological Sciences", title = "Glikozilacija i pH stabilnost penicilin G acilaze iz providencia rettgeri proizvedene u Pichia pastoris, Glycosylation and pH stability of penicillin G acylase from providencia rettgeri produced in Pichia pastoris", pages = "586-581", number = "4", volume = "61", doi = "10.2298/ABS0904581S" }
Šenerović, L., Stanković, N., Ljubijankić, G.,& Vasiljević, B.. (2009). Glikozilacija i pH stabilnost penicilin G acilaze iz providencia rettgeri proizvedene u Pichia pastoris. in Archives of Biological Sciences Srpsko biološko društvo, Beograd, i dr.., 61(4), 581-586. https://doi.org/10.2298/ABS0904581S
Šenerović L, Stanković N, Ljubijankić G, Vasiljević B. Glikozilacija i pH stabilnost penicilin G acilaze iz providencia rettgeri proizvedene u Pichia pastoris. in Archives of Biological Sciences. 2009;61(4):581-586. doi:10.2298/ABS0904581S .
Šenerović, Lidija, Stanković, Nada, Ljubijankić, G., Vasiljević, Branka, "Glikozilacija i pH stabilnost penicilin G acilaze iz providencia rettgeri proizvedene u Pichia pastoris" in Archives of Biological Sciences, 61, no. 4 (2009):581-586, https://doi.org/10.2298/ABS0904581S . .