TY  - 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 . .

TY  - JOUR
AU  - Šenerović, Lidija
AU  - Stanković, Nada
AU  - Spizzo, P
AU  - Basso, A
AU  - Gardossi, L
AU  - Vasiljević, Branka
AU  - Ljubijankić, G
AU  - Tisminetzky, S
AU  - Degrassi, G
PY  - 2006
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/255
AB  - A complete, integrated process for the production of an innovative formulation of penicillin G acylase from Providencia rettgeri (rPAC(P).(rett)) of industrial applicability is reported. In order to improve the yield of rPAC, the clone LN5.5, carrying four copies of pac gene integrated into the genome of Pichia pastoris, was constructed. The proteinase activity of the recombinant strain was reduced by knockout of the PEN gene encoding for proteinase A, resulting in an increased rPAC(P.rett) activity of approximately 40% (3.8 U/mL vs. 2.7 U/mL produced by LN5.5 in flask). A high cell density fermentation process was established with a 5-day methanol induction phase and a final PAC activity of up to 27 U/mL. A single step rPAC(P.rett) purification was also developed with an enzyme activity yield of approximately 95%. The novel features of the rPAC(P.rett) expressed in P. pastoris were fully exploited and emphasized through the covalent immobilization of rPAC(P.rett). The enzyme was immobilized on a series of structurally correlated methacrylic polymers, specifically designed and produced for optimizing rPAC(P.rett) performances in both hydrolytic and synthetic processes. Polymers presenting aminic functionalities were the most efficient, leading to formulations with higher activity and stability (half time stability  gt  3 years and specific activity ranging from 237 to 477 U/g (dry) based on benzylpenicillin hydrolysis). The efficiency of the immobilized rPAC(P.rett) was finally evaluated by studying the kinetically controlled synthesis of P-lactam antibiotics (cephalexin) and estimating the synthesis/hydrolysis ratio (S/H), which is a crucial parameter for the feasibility of the process.
PB  - Wiley, Hoboken
T2  - Biotechnology and Bioengineering
T1  - High-level production and covalent immobilization of Providencia rettgeri penicillin G acylase (PAC) from recombinant Pichia pastoris for the development of a novel and stable biocatalyst of industrial applicability
EP  - 354
IS  - 2
SP  - 344
VL  - 93
DO  - 10.1002/bit.20728
ER  - 

@article{
author = "Šenerović, Lidija and Stanković, Nada and Spizzo, P and Basso, A and Gardossi, L and Vasiljević, Branka and Ljubijankić, G and Tisminetzky, S and Degrassi, G",
year = "2006",
abstract = "A complete, integrated process for the production of an innovative formulation of penicillin G acylase from Providencia rettgeri (rPAC(P).(rett)) of industrial applicability is reported. In order to improve the yield of rPAC, the clone LN5.5, carrying four copies of pac gene integrated into the genome of Pichia pastoris, was constructed. The proteinase activity of the recombinant strain was reduced by knockout of the PEN gene encoding for proteinase A, resulting in an increased rPAC(P.rett) activity of approximately 40% (3.8 U/mL vs. 2.7 U/mL produced by LN5.5 in flask). A high cell density fermentation process was established with a 5-day methanol induction phase and a final PAC activity of up to 27 U/mL. A single step rPAC(P.rett) purification was also developed with an enzyme activity yield of approximately 95%. The novel features of the rPAC(P.rett) expressed in P. pastoris were fully exploited and emphasized through the covalent immobilization of rPAC(P.rett). The enzyme was immobilized on a series of structurally correlated methacrylic polymers, specifically designed and produced for optimizing rPAC(P.rett) performances in both hydrolytic and synthetic processes. Polymers presenting aminic functionalities were the most efficient, leading to formulations with higher activity and stability (half time stability  gt  3 years and specific activity ranging from 237 to 477 U/g (dry) based on benzylpenicillin hydrolysis). The efficiency of the immobilized rPAC(P.rett) was finally evaluated by studying the kinetically controlled synthesis of P-lactam antibiotics (cephalexin) and estimating the synthesis/hydrolysis ratio (S/H), which is a crucial parameter for the feasibility of the process.",
publisher = "Wiley, Hoboken",
journal = "Biotechnology and Bioengineering",
title = "High-level production and covalent immobilization of Providencia rettgeri penicillin G acylase (PAC) from recombinant Pichia pastoris for the development of a novel and stable biocatalyst of industrial applicability",
pages = "354-344",
number = "2",
volume = "93",
doi = "10.1002/bit.20728"
}

Šenerović, L., Stanković, N., Spizzo, P., Basso, A., Gardossi, L., Vasiljević, B., Ljubijankić, G., Tisminetzky, S.,& Degrassi, G.. (2006). High-level production and covalent immobilization of Providencia rettgeri penicillin G acylase (PAC) from recombinant Pichia pastoris for the development of a novel and stable biocatalyst of industrial applicability. in Biotechnology and Bioengineering
Wiley, Hoboken., 93(2), 344-354.
https://doi.org/10.1002/bit.20728

Šenerović L, Stanković N, Spizzo P, Basso A, Gardossi L, Vasiljević B, Ljubijankić G, Tisminetzky S, Degrassi G. High-level production and covalent immobilization of Providencia rettgeri penicillin G acylase (PAC) from recombinant Pichia pastoris for the development of a novel and stable biocatalyst of industrial applicability. in Biotechnology and Bioengineering. 2006;93(2):344-354.
doi:10.1002/bit.20728 .

Šenerović, Lidija, Stanković, Nada, Spizzo, P, Basso, A, Gardossi, L, Vasiljević, Branka, Ljubijankić, G, Tisminetzky, S, Degrassi, G, "High-level production and covalent immobilization of Providencia rettgeri penicillin G acylase (PAC) from recombinant Pichia pastoris for the development of a novel and stable biocatalyst of industrial applicability" in Biotechnology and Bioengineering, 93, no. 2 (2006):344-354,
https://doi.org/10.1002/bit.20728 . .

TY  - JOUR
AU  - Skoko, N
AU  - Argamante, B
AU  - Kovačević Grujičić, Nataša
AU  - Grujicić, NK
AU  - Tisminetzky, SG
AU  - Glisin, V
AU  - Ljubijankić, G
PY  - 2003
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/182
AB  - We describe the heterologous expression of a human interferon-beta1 in the methylotrophic yeast Pichia pastoris. Biologically active recombinant human interferon-beta1 (rHulFN-beta1) was secreted from shake-flask-grown P pastoris cells into the medium using the Saccharomyces cerevisiae alpha-mating factor prepro-leader sequence at the level of (1-3) x 10(5) i.u. (international units)/ml (6-12 mg/litre). An rHulFN-beta1 with an N-terminal sequence identical with that of native HulFN-beta1 was purified and the specific activity was determined (2-3 x 10(7) i.u./mg). It was found that the secreted recombinant protein was partially N-glycosylated.
PB  - Wiley, Hoboken
T2  - Biotechnology and Applied Biochemistry
T1  - Expression and characterization of human interferon-beta I in the methylotrophic yeast Pichia pastoris
EP  - 265
SP  - 257
VL  - 38
DO  - 10.1042/BA20030065
ER  - 

@article{
author = "Skoko, N and Argamante, B and Kovačević Grujičić, Nataša and Grujicić, NK and Tisminetzky, SG and Glisin, V and Ljubijankić, G",
year = "2003",
abstract = "We describe the heterologous expression of a human interferon-beta1 in the methylotrophic yeast Pichia pastoris. Biologically active recombinant human interferon-beta1 (rHulFN-beta1) was secreted from shake-flask-grown P pastoris cells into the medium using the Saccharomyces cerevisiae alpha-mating factor prepro-leader sequence at the level of (1-3) x 10(5) i.u. (international units)/ml (6-12 mg/litre). An rHulFN-beta1 with an N-terminal sequence identical with that of native HulFN-beta1 was purified and the specific activity was determined (2-3 x 10(7) i.u./mg). It was found that the secreted recombinant protein was partially N-glycosylated.",
publisher = "Wiley, Hoboken",
journal = "Biotechnology and Applied Biochemistry",
title = "Expression and characterization of human interferon-beta I in the methylotrophic yeast Pichia pastoris",
pages = "265-257",
volume = "38",
doi = "10.1042/BA20030065"
}

Skoko, N., Argamante, B., Kovačević Grujičić, N., Grujicić, N., Tisminetzky, S., Glisin, V.,& Ljubijankić, G.. (2003). Expression and characterization of human interferon-beta I in the methylotrophic yeast Pichia pastoris. in Biotechnology and Applied Biochemistry
Wiley, Hoboken., 38, 257-265.
https://doi.org/10.1042/BA20030065

Skoko N, Argamante B, Kovačević Grujičić N, Grujicić N, Tisminetzky S, Glisin V, Ljubijankić G. Expression and characterization of human interferon-beta I in the methylotrophic yeast Pichia pastoris. in Biotechnology and Applied Biochemistry. 2003;38:257-265.
doi:10.1042/BA20030065 .

Skoko, N, Argamante, B, Kovačević Grujičić, Nataša, Grujicić, NK, Tisminetzky, SG, Glisin, V, Ljubijankić, G, "Expression and characterization of human interferon-beta I in the methylotrophic yeast Pichia pastoris" in Biotechnology and Applied Biochemistry, 38 (2003):257-265,
https://doi.org/10.1042/BA20030065 . .

TY  - JOUR
AU  - Degrassi, G
AU  - Kojić, Milan
AU  - Ljubijankić, G
AU  - Venturi, V
PY  - 2000
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/140
AB  - The Bacillus pumilus gene encoding acetyl xylan esterase tare) was identified and characterized. The axe gene was expressed and the recombinant enzyme produced in Escherichia coli was purified and characterized. The recombinant enzyme displayed similar properties to the acetyl xylan esterase (AXE) from B. pumilus. The AXE primary structure was 76% identical to the cephalosporin C deacetylase of B. subtilis, and 40% to two recently identified AXEs from Thermoanaerobacterium and Thermotoga maritima. These four proteins are of similar sire and represent a new family of esterases having a broad substrate specificity. The recombinant AXE was demonstrated to have activity on several acetylated substrates, including on cephalosporin C.
PB  - Microbiology Soc, London
T2  - Microbiology-Sgm
T1  - The acetyl xylan esterase of Bacillus pumilus belongs to a family of esterases with broad substrate specificity
EP  - 1591
SP  - 1585
VL  - 146
DO  - 10.1099/00221287-146-7-1585
ER  - 

@article{
author = "Degrassi, G and Kojić, Milan and Ljubijankić, G and Venturi, V",
year = "2000",
abstract = "The Bacillus pumilus gene encoding acetyl xylan esterase tare) was identified and characterized. The axe gene was expressed and the recombinant enzyme produced in Escherichia coli was purified and characterized. The recombinant enzyme displayed similar properties to the acetyl xylan esterase (AXE) from B. pumilus. The AXE primary structure was 76% identical to the cephalosporin C deacetylase of B. subtilis, and 40% to two recently identified AXEs from Thermoanaerobacterium and Thermotoga maritima. These four proteins are of similar sire and represent a new family of esterases having a broad substrate specificity. The recombinant AXE was demonstrated to have activity on several acetylated substrates, including on cephalosporin C.",
publisher = "Microbiology Soc, London",
journal = "Microbiology-Sgm",
title = "The acetyl xylan esterase of Bacillus pumilus belongs to a family of esterases with broad substrate specificity",
pages = "1591-1585",
volume = "146",
doi = "10.1099/00221287-146-7-1585"
}

Degrassi, G., Kojić, M., Ljubijankić, G.,& Venturi, V.. (2000). The acetyl xylan esterase of Bacillus pumilus belongs to a family of esterases with broad substrate specificity. in Microbiology-Sgm
Microbiology Soc, London., 146, 1585-1591.
https://doi.org/10.1099/00221287-146-7-1585

Degrassi G, Kojić M, Ljubijankić G, Venturi V. The acetyl xylan esterase of Bacillus pumilus belongs to a family of esterases with broad substrate specificity. in Microbiology-Sgm. 2000;146:1585-1591.
doi:10.1099/00221287-146-7-1585 .

Degrassi, G, Kojić, Milan, Ljubijankić, G, Venturi, V, "The acetyl xylan esterase of Bacillus pumilus belongs to a family of esterases with broad substrate specificity" in Microbiology-Sgm, 146 (2000):1585-1591,
https://doi.org/10.1099/00221287-146-7-1585 . .