TY  - CONF
AU  - Stefanović, Marija
AU  - Savić, Aleksa
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Radosavljević, Jelena
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6206
AB  - Viral exonucleases play role in many processes essential for genome ma intenance,including DNA repair and recombination. Lambda exonuclease (λ -exo), isolated fromlambda bacteriophage, hydrolases double-stranded DNA (dsDNA) in the highly processivemanner in 5’→3’ direction, yielding mononucleotides and single -stranded DNA (ssDNA).This unique enzymatic properties offer several promising biotechnological applications,such as highly sensitive quantification of DNA modifications and single -moleculesequencing. Hence, optimization of the expression conditions is a prerequisite to achievehigh-level production of λ-exo. Here we have tested λ -exo expression in five different E.coli strains under various temperature regimes in order to establish the optimal conditionsfor efficient production of recombinant λ -exo. The N-terminally His -tagged λ-exo wassuccessfully expressed in E.coli BL21(AI), SHuffle T7, C41(DE3) and C43(DE3) strains inLB broth. Collected aliquots were analysed by SDS-PAGE, followed by CBB staining.Relative yield of target protein bands was determined by densitometry in total cell lysate, aswell as in soluble and insoluble cytoplasmatic fractions. We identified E.coli BL21(AI),SHuffle T7 and C41(DE3) as good producers of recombinant λ -exo, and upon scaling up, λ-exo was purified from crude cell lysates by metal affinity chromatography in satisfactoryyield. Our data suggest that densitometric analysis could serve as a powerful low-costscreening platform for improving recombinant protein expression strategies.
PB  - Prirodno-matematički fakultet, Univerzitet u Kragujevcu
C3  - Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac
T1  - Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6206
ER  - 

@conference{
author = "Stefanović, Marija and Savić, Aleksa and Božić, Nataša and Vujčić, Zoran and Radosavljević, Jelena",
year = "2023",
abstract = "Viral exonucleases play role in many processes essential for genome ma intenance,including DNA repair and recombination. Lambda exonuclease (λ -exo), isolated fromlambda bacteriophage, hydrolases double-stranded DNA (dsDNA) in the highly processivemanner in 5’→3’ direction, yielding mononucleotides and single -stranded DNA (ssDNA).This unique enzymatic properties offer several promising biotechnological applications,such as highly sensitive quantification of DNA modifications and single -moleculesequencing. Hence, optimization of the expression conditions is a prerequisite to achievehigh-level production of λ-exo. Here we have tested λ -exo expression in five different E.coli strains under various temperature regimes in order to establish the optimal conditionsfor efficient production of recombinant λ -exo. The N-terminally His -tagged λ-exo wassuccessfully expressed in E.coli BL21(AI), SHuffle T7, C41(DE3) and C43(DE3) strains inLB broth. Collected aliquots were analysed by SDS-PAGE, followed by CBB staining.Relative yield of target protein bands was determined by densitometry in total cell lysate, aswell as in soluble and insoluble cytoplasmatic fractions. We identified E.coli BL21(AI),SHuffle T7 and C41(DE3) as good producers of recombinant λ -exo, and upon scaling up, λ-exo was purified from crude cell lysates by metal affinity chromatography in satisfactoryyield. Our data suggest that densitometric analysis could serve as a powerful low-costscreening platform for improving recombinant protein expression strategies.",
publisher = "Prirodno-matematički fakultet, Univerzitet u Kragujevcu",
journal = "Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac",
title = "Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6206"
}

Stefanović, M., Savić, A., Božić, N., Vujčić, Z.,& Radosavljević, J.. (2023). Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains. in Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac
Prirodno-matematički fakultet, Univerzitet u Kragujevcu..
https://hdl.handle.net/21.15107/rcub_cherry_6206

Stefanović M, Savić A, Božić N, Vujčić Z, Radosavljević J. Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains. in Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac. 2023;.
https://hdl.handle.net/21.15107/rcub_cherry_6206 .

Stefanović, Marija, Savić, Aleksa, Božić, Nataša, Vujčić, Zoran, Radosavljević, Jelena, "Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains" in Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac (2023),
https://hdl.handle.net/21.15107/rcub_cherry_6206 .

TY  - CONF
AU  - Stefanović, Marija
AU  - Savić, Aleksa
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Radosavljević, Jelena
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6205
AB  - Viral exonucleases play role in many processes essential for genome ma intenance,
including DNA repair and recombination. Lambda exonuclease (λ -exo), isolated from
lambda bacteriophage, hydrolases double-stranded DNA (dsDNA) in the highly processive
manner in 5’→3’ direction, yielding mononucleotides and single -stranded DNA (ssDNA).
This unique enzymatic properties offer several promising biotechnological applications,
such as highly sensitive quantification of DNA modifications and single -molecule
sequencing. Hence, optimization of the expression conditions is a prerequisite to achieve
high-level production of λ-exo. Here we have tested λ -exo expression in five different E.
coli strains under various temperature regimes in order to establish the optimal conditions
for efficient production of recombinant λ -exo. The N-terminally His -tagged λ-exo was
successfully expressed in E.coli BL21(AI), SHuffle T7, C41(DE3) and C43(DE3) strains in
LB broth. Collected aliquots were analysed by SDS-PAGE, followed by CBB staining.
Relative yield of target protein bands was determined by densitometry in total cell lysate, as
well as in soluble and insoluble cytoplasmatic fractions. We identified E.coli BL21(AI),
SHuffle T7 and C41(DE3) as good producers of recombinant λ -exo, and upon scaling up, λ
-exo was purified from crude cell lysates by metal affinity chromatography in satisfactory
yield. Our data suggest that densitometric analysis could serve as a powerful low-cost
screening platform for improving recombinant protein expression strategies.
PB  - Prirodno-matematički fakultet, Univerzitet u Kragujevcu
C3  - Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac
T1  - Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains
SP  - 23
EP  - 23
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6205
ER  - 

@conference{
author = "Stefanović, Marija and Savić, Aleksa and Božić, Nataša and Vujčić, Zoran and Radosavljević, Jelena",
year = "2023",
abstract = "Viral exonucleases play role in many processes essential for genome ma intenance,
including DNA repair and recombination. Lambda exonuclease (λ -exo), isolated from
lambda bacteriophage, hydrolases double-stranded DNA (dsDNA) in the highly processive
manner in 5’→3’ direction, yielding mononucleotides and single -stranded DNA (ssDNA).
This unique enzymatic properties offer several promising biotechnological applications,
such as highly sensitive quantification of DNA modifications and single -molecule
sequencing. Hence, optimization of the expression conditions is a prerequisite to achieve
high-level production of λ-exo. Here we have tested λ -exo expression in five different E.
coli strains under various temperature regimes in order to establish the optimal conditions
for efficient production of recombinant λ -exo. The N-terminally His -tagged λ-exo was
successfully expressed in E.coli BL21(AI), SHuffle T7, C41(DE3) and C43(DE3) strains in
LB broth. Collected aliquots were analysed by SDS-PAGE, followed by CBB staining.
Relative yield of target protein bands was determined by densitometry in total cell lysate, as
well as in soluble and insoluble cytoplasmatic fractions. We identified E.coli BL21(AI),
SHuffle T7 and C41(DE3) as good producers of recombinant λ -exo, and upon scaling up, λ
-exo was purified from crude cell lysates by metal affinity chromatography in satisfactory
yield. Our data suggest that densitometric analysis could serve as a powerful low-cost
screening platform for improving recombinant protein expression strategies.",
publisher = "Prirodno-matematički fakultet, Univerzitet u Kragujevcu",
journal = "Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac",
title = "Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains",
pages = "23-23",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6205"
}

Stefanović, M., Savić, A., Božić, N., Vujčić, Z.,& Radosavljević, J.. (2023). Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains. in Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac
Prirodno-matematički fakultet, Univerzitet u Kragujevcu., 23-23.
https://hdl.handle.net/21.15107/rcub_cherry_6205

Stefanović M, Savić A, Božić N, Vujčić Z, Radosavljević J. Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains. in Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac. 2023;:23-23.
https://hdl.handle.net/21.15107/rcub_cherry_6205 .

Stefanović, Marija, Savić, Aleksa, Božić, Nataša, Vujčić, Zoran, Radosavljević, Jelena, "Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains" in Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac (2023):23-23,
https://hdl.handle.net/21.15107/rcub_cherry_6205 .

TY  - JOUR
AU  - Slavić, Marinela Šokarda
AU  - Kojić, Milan
AU  - Margetić, Aleksandra
AU  - Stanisavljević, Nemanja S.
AU  - Gardijan, Lazar
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6351
AB  - α-Amylase from the thermophilic bacterial strain Anoxybacillus vranjensis ST4 (AVA) was cloned into the pMALc5HisEk expression vector and successfully expressed and purified from the Escherichia coli ER2523 host strain. AVA belongs to the GH13_5 subfamily of glycoside hydrolases and has 7 conserved sequence regions (CSRs) distributed in three distinct domains (A, B, C). In addition, there is a starch binding domain (SBD) from the CBM20 family of carbohydrate binding modules (CBMs). AVA is a monomer of 66 kDa that achieves maximum activity at 60–80 °C and is active and stable over a wide pH range (4.0–9.0). AVA retained 50 % of its activity after 31 h of incubation at 60 °C and was resistant to a large number of denaturing agents. It hydrolyzed starch granules very efficiently, releasing maltose, maltotriose and maltopentaose as the main products. The hydrolysis rates of raw corn, wheat, horseradish, and potato starch, at a concentration of 10 %, were 87.8, 85.9, 93.0, and 58 %, respectively, at pH 8.5 over a 3 h period. This study showed that the high level of expression as well as the properties of this highly stable and versatile enzyme show all the prerequisites for successful application in industry.
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T1  - Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications
VL  - 249
SP  - 126055
DO  - 10.1016/j.ijbiomac.2023.126055
ER  - 

@article{
author = "Slavić, Marinela Šokarda and Kojić, Milan and Margetić, Aleksandra and Stanisavljević, Nemanja S. and Gardijan, Lazar and Božić, Nataša and Vujčić, Zoran",
year = "2023",
abstract = "α-Amylase from the thermophilic bacterial strain Anoxybacillus vranjensis ST4 (AVA) was cloned into the pMALc5HisEk expression vector and successfully expressed and purified from the Escherichia coli ER2523 host strain. AVA belongs to the GH13_5 subfamily of glycoside hydrolases and has 7 conserved sequence regions (CSRs) distributed in three distinct domains (A, B, C). In addition, there is a starch binding domain (SBD) from the CBM20 family of carbohydrate binding modules (CBMs). AVA is a monomer of 66 kDa that achieves maximum activity at 60–80 °C and is active and stable over a wide pH range (4.0–9.0). AVA retained 50 % of its activity after 31 h of incubation at 60 °C and was resistant to a large number of denaturing agents. It hydrolyzed starch granules very efficiently, releasing maltose, maltotriose and maltopentaose as the main products. The hydrolysis rates of raw corn, wheat, horseradish, and potato starch, at a concentration of 10 %, were 87.8, 85.9, 93.0, and 58 %, respectively, at pH 8.5 over a 3 h period. This study showed that the high level of expression as well as the properties of this highly stable and versatile enzyme show all the prerequisites for successful application in industry.",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules",
title = "Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications",
volume = "249",
pages = "126055",
doi = "10.1016/j.ijbiomac.2023.126055"
}

Slavić, M. Š., Kojić, M., Margetić, A., Stanisavljević, N. S., Gardijan, L., Božić, N.,& Vujčić, Z.. (2023). Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications. in International Journal of Biological Macromolecules
Elsevier., 249, 126055.
https://doi.org/10.1016/j.ijbiomac.2023.126055

Slavić MŠ, Kojić M, Margetić A, Stanisavljević NS, Gardijan L, Božić N, Vujčić Z. Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications. in International Journal of Biological Macromolecules. 2023;249:126055.
doi:10.1016/j.ijbiomac.2023.126055 .

Slavić, Marinela Šokarda, Kojić, Milan, Margetić, Aleksandra, Stanisavljević, Nemanja S., Gardijan, Lazar, Božić, Nataša, Vujčić, Zoran, "Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications" in International Journal of Biological Macromolecules, 249 (2023):126055,
https://doi.org/10.1016/j.ijbiomac.2023.126055 . .

TY  - JOUR
AU  - Šokarda Slavić, Marinela
AU  - Margetić, Aleksandra
AU  - Dojnov, Biljana
AU  - Vujčić, Miroslava
AU  - Mišić, Milan
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5796
AB  - Bioethanol is one of the main bio-based molecules produced mainly from sugar cane, molasses and corn. Its environmental advantages allow it to be considered as safe and the cleanest fuel alternative. Starch is a widespread renewable carbohydrate conventionally used for bioethanol production via energy demanding liquefaction and saccharification processes. Raw starch hydrolysis using enzymes capable of degrading it below the gelatinization temperature significantly simplifies the process and reduces the cost of starch processing. In this study, an innovative modified simultaneous saccharification and fermentation process is proposed for the production of bioethanol from highly concentrated raw corn starch (30 % w/v). A two-step synergistic hydrolysis and fermentation was carried out in a single bioreactor vessel. To ensure high process efficiency, factors influencing the hydrolysis of concentrated raw corn starch by raw starch degrading α-amylase from Bacillus paralicheniformis ATCC 9945a (BliAmy) and commercial glucoamylase were investigated. Box–Behnken experimental design was used to predict the effects of different ratios of added enzymes, glucoamylase addition time, incubation time, and pH on hydrolysis yield. Optimal conditions for the highest yield of hydrolysis of raw corn starch (90 %) were obtained after 8 h using 5.0 IU BliAmy per mg of starch and 0.5 % (v/v) glucoamylase at pH 4.5 and 60 °C. Obtained glucose was further fermented with Saccharomyces cerevisiae at 30 °C in the same vessel for bioethanol production. Bioethanol concentration at 129.2 g/L, with productivity of 2.94 g/L/h and ethanol yield (YP/S) at 0.50 g EtOH/g total sugar, equivalent to 87.8 % theoretical yield, was obtained by modified simultaneous saccharification and fermentation. This work enriches the information of bioethanol production and offers a novel strategy for raw starch hydrolysis under industrial conditions.
T2  - Fuel
T1  - Modified simultaneous saccharification and fermentation for the production of bioethanol from highly concentrated raw corn starch
VL  - 338
SP  - 127363
DO  - 10.1016/j.fuel.2022.127363
ER  - 

@article{
author = "Šokarda Slavić, Marinela and Margetić, Aleksandra and Dojnov, Biljana and Vujčić, Miroslava and Mišić, Milan and Božić, Nataša and Vujčić, Zoran",
year = "2023",
abstract = "Bioethanol is one of the main bio-based molecules produced mainly from sugar cane, molasses and corn. Its environmental advantages allow it to be considered as safe and the cleanest fuel alternative. Starch is a widespread renewable carbohydrate conventionally used for bioethanol production via energy demanding liquefaction and saccharification processes. Raw starch hydrolysis using enzymes capable of degrading it below the gelatinization temperature significantly simplifies the process and reduces the cost of starch processing. In this study, an innovative modified simultaneous saccharification and fermentation process is proposed for the production of bioethanol from highly concentrated raw corn starch (30 % w/v). A two-step synergistic hydrolysis and fermentation was carried out in a single bioreactor vessel. To ensure high process efficiency, factors influencing the hydrolysis of concentrated raw corn starch by raw starch degrading α-amylase from Bacillus paralicheniformis ATCC 9945a (BliAmy) and commercial glucoamylase were investigated. Box–Behnken experimental design was used to predict the effects of different ratios of added enzymes, glucoamylase addition time, incubation time, and pH on hydrolysis yield. Optimal conditions for the highest yield of hydrolysis of raw corn starch (90 %) were obtained after 8 h using 5.0 IU BliAmy per mg of starch and 0.5 % (v/v) glucoamylase at pH 4.5 and 60 °C. Obtained glucose was further fermented with Saccharomyces cerevisiae at 30 °C in the same vessel for bioethanol production. Bioethanol concentration at 129.2 g/L, with productivity of 2.94 g/L/h and ethanol yield (YP/S) at 0.50 g EtOH/g total sugar, equivalent to 87.8 % theoretical yield, was obtained by modified simultaneous saccharification and fermentation. This work enriches the information of bioethanol production and offers a novel strategy for raw starch hydrolysis under industrial conditions.",
journal = "Fuel",
title = "Modified simultaneous saccharification and fermentation for the production of bioethanol from highly concentrated raw corn starch",
volume = "338",
pages = "127363",
doi = "10.1016/j.fuel.2022.127363"
}

Šokarda Slavić, M., Margetić, A., Dojnov, B., Vujčić, M., Mišić, M., Božić, N.,& Vujčić, Z.. (2023). Modified simultaneous saccharification and fermentation for the production of bioethanol from highly concentrated raw corn starch. in Fuel, 338, 127363.
https://doi.org/10.1016/j.fuel.2022.127363

Šokarda Slavić M, Margetić A, Dojnov B, Vujčić M, Mišić M, Božić N, Vujčić Z. Modified simultaneous saccharification and fermentation for the production of bioethanol from highly concentrated raw corn starch. in Fuel. 2023;338:127363.
doi:10.1016/j.fuel.2022.127363 .

Šokarda Slavić, Marinela, Margetić, Aleksandra, Dojnov, Biljana, Vujčić, Miroslava, Mišić, Milan, Božić, Nataša, Vujčić, Zoran, "Modified simultaneous saccharification and fermentation for the production of bioethanol from highly concentrated raw corn starch" in Fuel, 338 (2023):127363,
https://doi.org/10.1016/j.fuel.2022.127363 . .

TY  - DATA
AU  - Šokarda Slavić, Marinela
AU  - Margetić, Aleksandra
AU  - Dojnov, Biljana
AU  - Vujčić, Miroslava
AU  - Mišić, Milan
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5836
AB  - Bioethanol is one of the main bio-based molecules produced mainly from sugar cane, molasses and corn. Its environmental advantages allow it to be considered as safe and the cleanest fuel alternative. Starch is a widespread renewable carbohydrate conventionally used for bioethanol production via energy demanding liquefaction and saccharification processes. Raw starch hydrolysis using enzymes capable of degrading it below the gelatinization temperature significantly simplifies the process and reduces the cost of starch processing. In this study, an innovative modified simultaneous saccharification and fermentation process is proposed for the production of bioethanol from highly concentrated raw corn starch (30 % w/v). A two-step synergistic hydrolysis and fermentation was carried out in a single bioreactor vessel. To ensure high process efficiency, factors influencing the hydrolysis of concentrated raw corn starch by raw starch degrading α-amylase from Bacillus paralicheniformis ATCC 9945a (BliAmy) and commercial glucoamylase were investigated. Box–Behnken experimental design was used to predict the effects of different ratios of added enzymes, glucoamylase addition time, incubation time, and pH on hydrolysis yield. Optimal conditions for the highest yield of hydrolysis of raw corn starch (90 %) were obtained after 8 h using 5.0 IU BliAmy per mg of starch and 0.5 % (v/v) glucoamylase at pH 4.5 and 60 °C. Obtained glucose was further fermented with Saccharomyces cerevisiae at 30 °C in the same vessel for bioethanol production. Bioethanol concentration at 129.2 g/L, with productivity of 2.94 g/L/h and ethanol yield (YP/S) at 0.50 g EtOH/g total sugar, equivalent to 87.8 % theoretical yield, was obtained by modified simultaneous saccharification and fermentation. This work enriches the information of bioethanol production and offers a novel strategy for raw starch hydrolysis under industrial conditions.
T2  - Fuel
T1  - Supplementary material for: Šokarda Slavić, M., Margetić, A., Dojnov, B., Vujčić, M., Mišić, M., Božić, N.,& Vujčić, Z.. (2023). Modified simultaneous saccharification and fermentation for the production of bioethanol from highly concentrated raw corn starch. in Fuel, 338, 127363. https://doi.org/10.1016/j.fuel.2022.127363
VL  - 338
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5836
ER  - 

@misc{
author = "Šokarda Slavić, Marinela and Margetić, Aleksandra and Dojnov, Biljana and Vujčić, Miroslava and Mišić, Milan and Božić, Nataša and Vujčić, Zoran",
year = "2023",
abstract = "Bioethanol is one of the main bio-based molecules produced mainly from sugar cane, molasses and corn. Its environmental advantages allow it to be considered as safe and the cleanest fuel alternative. Starch is a widespread renewable carbohydrate conventionally used for bioethanol production via energy demanding liquefaction and saccharification processes. Raw starch hydrolysis using enzymes capable of degrading it below the gelatinization temperature significantly simplifies the process and reduces the cost of starch processing. In this study, an innovative modified simultaneous saccharification and fermentation process is proposed for the production of bioethanol from highly concentrated raw corn starch (30 % w/v). A two-step synergistic hydrolysis and fermentation was carried out in a single bioreactor vessel. To ensure high process efficiency, factors influencing the hydrolysis of concentrated raw corn starch by raw starch degrading α-amylase from Bacillus paralicheniformis ATCC 9945a (BliAmy) and commercial glucoamylase were investigated. Box–Behnken experimental design was used to predict the effects of different ratios of added enzymes, glucoamylase addition time, incubation time, and pH on hydrolysis yield. Optimal conditions for the highest yield of hydrolysis of raw corn starch (90 %) were obtained after 8 h using 5.0 IU BliAmy per mg of starch and 0.5 % (v/v) glucoamylase at pH 4.5 and 60 °C. Obtained glucose was further fermented with Saccharomyces cerevisiae at 30 °C in the same vessel for bioethanol production. Bioethanol concentration at 129.2 g/L, with productivity of 2.94 g/L/h and ethanol yield (YP/S) at 0.50 g EtOH/g total sugar, equivalent to 87.8 % theoretical yield, was obtained by modified simultaneous saccharification and fermentation. This work enriches the information of bioethanol production and offers a novel strategy for raw starch hydrolysis under industrial conditions.",
journal = "Fuel",
title = "Supplementary material for: Šokarda Slavić, M., Margetić, A., Dojnov, B., Vujčić, M., Mišić, M., Božić, N.,& Vujčić, Z.. (2023). Modified simultaneous saccharification and fermentation for the production of bioethanol from highly concentrated raw corn starch. in Fuel, 338, 127363. https://doi.org/10.1016/j.fuel.2022.127363",
volume = "338",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5836"
}

Šokarda Slavić, M., Margetić, A., Dojnov, B., Vujčić, M., Mišić, M., Božić, N.,& Vujčić, Z.. (2023). Supplementary material for: Šokarda Slavić, M., Margetić, A., Dojnov, B., Vujčić, M., Mišić, M., Božić, N.,& Vujčić, Z.. (2023). Modified simultaneous saccharification and fermentation for the production of bioethanol from highly concentrated raw corn starch. in Fuel, 338, 127363. https://doi.org/10.1016/j.fuel.2022.127363. in Fuel, 338.
https://hdl.handle.net/21.15107/rcub_cherry_5836

Šokarda Slavić M, Margetić A, Dojnov B, Vujčić M, Mišić M, Božić N, Vujčić Z. Supplementary material for: Šokarda Slavić, M., Margetić, A., Dojnov, B., Vujčić, M., Mišić, M., Božić, N.,& Vujčić, Z.. (2023). Modified simultaneous saccharification and fermentation for the production of bioethanol from highly concentrated raw corn starch. in Fuel, 338, 127363. https://doi.org/10.1016/j.fuel.2022.127363. in Fuel. 2023;338.
https://hdl.handle.net/21.15107/rcub_cherry_5836 .

Šokarda Slavić, Marinela, Margetić, Aleksandra, Dojnov, Biljana, Vujčić, Miroslava, Mišić, Milan, Božić, Nataša, Vujčić, Zoran, "Supplementary material for: Šokarda Slavić, M., Margetić, A., Dojnov, B., Vujčić, M., Mišić, M., Božić, N.,& Vujčić, Z.. (2023). Modified simultaneous saccharification and fermentation for the production of bioethanol from highly concentrated raw corn starch. in Fuel, 338, 127363. https://doi.org/10.1016/j.fuel.2022.127363" in Fuel, 338 (2023),
https://hdl.handle.net/21.15107/rcub_cherry_5836 .

TY  - CONF
AU  - Stefanović, Marija
AU  - Savić, Aleksa
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Radosavljević, Jelena
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6207
AB  - Introduction: Lambda exonuclease (λ-exo), isolated from lambda bacteriophage, plays a crucial role in
DNA replication, repair and recombination. The enzyme specifically hydrolases double-stranded DNA
(dsDNA) in a highly processive manner in 5’�����3’ direction, yielding mononucleotides and single-stranded
DNA (ssDNA). This efficient unidirectional degradation makes it an invaluable tool in various molecular
biology techniques, including novel sequencing technologies. Hence, optimization of the expression
conditions is a prerequisite to achieving high-level production of λ-exo.
Methods: The N-terminally His-tagged λ-exo fusion construct with thrombin cleavage site (AddGene
#104531) was successfully transformed into five different E. coli strains (BL21(AI), Shuffle T7, C41(DE3),
C43(DE3), and BL21(DE3)). Expression was tested under three temperature regimes (20 ˚C, 30 ˚C, and 37
˚C) over time for enhanced soluble production. Crude extracts were analysed by SDS-PAGE for total protein
expression, soluble and insoluble cytoplasmatic fractions. The exonuclease activity of the extracts
was monitored via in-house developed fluorescence-based screening assay. Optimal conditions for highyield
production were determined by densitometric analysis using NIH ImageJ software. The soluble and
active enzyme was produced on the large scale in a shaking flask culture under optimal conditions, and
purified to homogeneity from the soluble lysate via metal affinity chromatography.
Results:We identified E. coli BL21(AI), SHuffle T7, and C41(DE3) as good producers of recombinant λ-exo
and determined optimal conditions (30 ˚C, 6 h post-induction) for high-yield expression. The enzyme was
eluted from Ni2+-IDA-Sepharose 6B column in 300 mM imidazole and maintained its activity upon purification
assessed by an in-house developed fluorescence-based screening assay.
Conclusion: This study provides a scalable cost-effective approach for soluble λ-exo production in selected
E. coli strains. This expression system would be a helpful platform for development of high-yield
production of λ-exo, easing its exploitation in biotechnology and other scientific frontiers. Additionally,
we provided a valuable low-cost screening assay for monitoring exonuclease activity during each purification
step.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue
T1  - Recombinant production of native λ-exonuclease in different E. coli strains
SP  - 172
EP  - 172
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6207
ER  - 

@conference{
author = "Stefanović, Marija and Savić, Aleksa and Božić, Nataša and Vujčić, Zoran and Radosavljević, Jelena",
year = "2023",
abstract = "Introduction: Lambda exonuclease (λ-exo), isolated from lambda bacteriophage, plays a crucial role in
DNA replication, repair and recombination. The enzyme specifically hydrolases double-stranded DNA
(dsDNA) in a highly processive manner in 5’�����3’ direction, yielding mononucleotides and single-stranded
DNA (ssDNA). This efficient unidirectional degradation makes it an invaluable tool in various molecular
biology techniques, including novel sequencing technologies. Hence, optimization of the expression
conditions is a prerequisite to achieving high-level production of λ-exo.
Methods: The N-terminally His-tagged λ-exo fusion construct with thrombin cleavage site (AddGene
#104531) was successfully transformed into five different E. coli strains (BL21(AI), Shuffle T7, C41(DE3),
C43(DE3), and BL21(DE3)). Expression was tested under three temperature regimes (20 ˚C, 30 ˚C, and 37
˚C) over time for enhanced soluble production. Crude extracts were analysed by SDS-PAGE for total protein
expression, soluble and insoluble cytoplasmatic fractions. The exonuclease activity of the extracts
was monitored via in-house developed fluorescence-based screening assay. Optimal conditions for highyield
production were determined by densitometric analysis using NIH ImageJ software. The soluble and
active enzyme was produced on the large scale in a shaking flask culture under optimal conditions, and
purified to homogeneity from the soluble lysate via metal affinity chromatography.
Results:We identified E. coli BL21(AI), SHuffle T7, and C41(DE3) as good producers of recombinant λ-exo
and determined optimal conditions (30 ˚C, 6 h post-induction) for high-yield expression. The enzyme was
eluted from Ni2+-IDA-Sepharose 6B column in 300 mM imidazole and maintained its activity upon purification
assessed by an in-house developed fluorescence-based screening assay.
Conclusion: This study provides a scalable cost-effective approach for soluble λ-exo production in selected
E. coli strains. This expression system would be a helpful platform for development of high-yield
production of λ-exo, easing its exploitation in biotechnology and other scientific frontiers. Additionally,
we provided a valuable low-cost screening assay for monitoring exonuclease activity during each purification
step.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue",
title = "Recombinant production of native λ-exonuclease in different E. coli strains",
pages = "172-172",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6207"
}

Stefanović, M., Savić, A., Božić, N., Vujčić, Z.,& Radosavljević, J.. (2023). Recombinant production of native λ-exonuclease in different E. coli strains. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 172-172.
https://hdl.handle.net/21.15107/rcub_cherry_6207

Stefanović M, Savić A, Božić N, Vujčić Z, Radosavljević J. Recombinant production of native λ-exonuclease in different E. coli strains. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue. 2023;:172-172.
https://hdl.handle.net/21.15107/rcub_cherry_6207 .

Stefanović, Marija, Savić, Aleksa, Božić, Nataša, Vujčić, Zoran, Radosavljević, Jelena, "Recombinant production of native λ-exonuclease in different E. coli strains" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue (2023):172-172,
https://hdl.handle.net/21.15107/rcub_cherry_6207 .

TY  - CONF
AU  - Stefanović, Marija
AU  - Savić, Aleksa
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Radosavljević, Jelena
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6208
AB  - Introduction: Lambda exonuclease (λ-exo), isolated from lambda bacteriophage, plays a crucial role inDNA replication, repair and recombination. The enzyme specifically hydrolases double-stranded DNA(dsDNA) in a highly processive manner in 5’�����3’ direction, yielding mononucleotides and single-strandedDNA (ssDNA). This efficient unidirectional degradation makes it an invaluable tool in various molecularbiology techniques, including novel sequencing technologies. Hence, optimization of the expressionconditions is a prerequisite to achieving high-level production of λ-exo.Methods: The N-terminally His-tagged λ-exo fusion construct with thrombin cleavage site (AddGene#104531) was successfully transformed into five different E. coli strains (BL21(AI), Shuffle T7, C41(DE3),C43(DE3), and BL21(DE3)). Expression was tested under three temperature regimes (20 ˚C, 30 ˚C, and 37˚C) over time for enhanced soluble production. Crude extracts were analysed by SDS-PAGE for total proteinexpression, soluble and insoluble cytoplasmatic fractions. The exonuclease activity of the extractswas monitored via in-house developed fluorescence-based screening assay. Optimal conditions for highyieldproduction were determined by densitometric analysis using NIH ImageJ software. The soluble andactive enzyme was produced on the large scale in a shaking flask culture under optimal conditions, andpurified to homogeneity from the soluble lysate via metal affinity chromatography.Results:We identified E. coli BL21(AI), SHuffle T7, and C41(DE3) as good producers of recombinant λ-exoand determined optimal conditions (30 ˚C, 6 h post-induction) for high-yield expression. The enzyme waseluted from Ni2+-IDA-Sepharose 6B column in 300 mM imidazole and maintained its activity upon purificationassessed by an in-house developed fluorescence-based screening assay.Conclusion: This study provides a scalable cost-effective approach for soluble λ-exo production in selectedE. coli strains. This expression system would be a helpful platform for development of high-yieldproduction of λ-exo, easing its exploitation in biotechnology and other scientific frontiers. Additionally,we provided a valuable low-cost screening assay for monitoring exonuclease activity during each purificationstep.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue
T1  - Recombinant production of native λ-exonuclease in different E. coli strains
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6208
ER  - 

@conference{
author = "Stefanović, Marija and Savić, Aleksa and Božić, Nataša and Vujčić, Zoran and Radosavljević, Jelena",
year = "2023",
abstract = "Introduction: Lambda exonuclease (λ-exo), isolated from lambda bacteriophage, plays a crucial role inDNA replication, repair and recombination. The enzyme specifically hydrolases double-stranded DNA(dsDNA) in a highly processive manner in 5’�����3’ direction, yielding mononucleotides and single-strandedDNA (ssDNA). This efficient unidirectional degradation makes it an invaluable tool in various molecularbiology techniques, including novel sequencing technologies. Hence, optimization of the expressionconditions is a prerequisite to achieving high-level production of λ-exo.Methods: The N-terminally His-tagged λ-exo fusion construct with thrombin cleavage site (AddGene#104531) was successfully transformed into five different E. coli strains (BL21(AI), Shuffle T7, C41(DE3),C43(DE3), and BL21(DE3)). Expression was tested under three temperature regimes (20 ˚C, 30 ˚C, and 37˚C) over time for enhanced soluble production. Crude extracts were analysed by SDS-PAGE for total proteinexpression, soluble and insoluble cytoplasmatic fractions. The exonuclease activity of the extractswas monitored via in-house developed fluorescence-based screening assay. Optimal conditions for highyieldproduction were determined by densitometric analysis using NIH ImageJ software. The soluble andactive enzyme was produced on the large scale in a shaking flask culture under optimal conditions, andpurified to homogeneity from the soluble lysate via metal affinity chromatography.Results:We identified E. coli BL21(AI), SHuffle T7, and C41(DE3) as good producers of recombinant λ-exoand determined optimal conditions (30 ˚C, 6 h post-induction) for high-yield expression. The enzyme waseluted from Ni2+-IDA-Sepharose 6B column in 300 mM imidazole and maintained its activity upon purificationassessed by an in-house developed fluorescence-based screening assay.Conclusion: This study provides a scalable cost-effective approach for soluble λ-exo production in selectedE. coli strains. This expression system would be a helpful platform for development of high-yieldproduction of λ-exo, easing its exploitation in biotechnology and other scientific frontiers. Additionally,we provided a valuable low-cost screening assay for monitoring exonuclease activity during each purificationstep.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue",
title = "Recombinant production of native λ-exonuclease in different E. coli strains",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6208"
}

Stefanović, M., Savić, A., Božić, N., Vujčić, Z.,& Radosavljević, J.. (2023). Recombinant production of native λ-exonuclease in different E. coli strains. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade..
https://hdl.handle.net/21.15107/rcub_cherry_6208

Stefanović M, Savić A, Božić N, Vujčić Z, Radosavljević J. Recombinant production of native λ-exonuclease in different E. coli strains. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue. 2023;.
https://hdl.handle.net/21.15107/rcub_cherry_6208 .

Stefanović, Marija, Savić, Aleksa, Božić, Nataša, Vujčić, Zoran, Radosavljević, Jelena, "Recombinant production of native λ-exonuclease in different E. coli strains" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue (2023),
https://hdl.handle.net/21.15107/rcub_cherry_6208 .

TY  - DATA
AU  - Simić, Stefan
AU  - Jeremić, Sanja
AU  - Đokić, Lidija
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Lončar, Nikola L.
AU  - Senthamaraikannan, Ramsankar
AU  - Babu, Ramesh P.
AU  - Opsenica, Igor
AU  - Nikodinović-Runić, Jasmina
PY  - 2020
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3357
T2  - Enzyme and Microbial Technology
T1  - Supplementary data for article: Simić, S.; Jeremic, S.; Djokic, L.; Božić, N.; Vujčić, Z.; Lončar, N.; Senthamaraikannan, R.; Babu, R.; Opsenica, I. M.; Nikodinovic-Runic, J. Development of an Efficient Biocatalytic System Based on Bacterial Laccase for the Oxidation of Selected 1,4-Dihydropyridines. Enzyme and Microbial Technology 2020, 132. https://doi.org/10.1016/j.enzmictec.2019.109411
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3357
ER  - 

@misc{
author = "Simić, Stefan and Jeremić, Sanja and Đokić, Lidija and Božić, Nataša and Vujčić, Zoran and Lončar, Nikola L. and Senthamaraikannan, Ramsankar and Babu, Ramesh P. and Opsenica, Igor and Nikodinović-Runić, Jasmina",
year = "2020",
journal = "Enzyme and Microbial Technology",
title = "Supplementary data for article: Simić, S.; Jeremic, S.; Djokic, L.; Božić, N.; Vujčić, Z.; Lončar, N.; Senthamaraikannan, R.; Babu, R.; Opsenica, I. M.; Nikodinovic-Runic, J. Development of an Efficient Biocatalytic System Based on Bacterial Laccase for the Oxidation of Selected 1,4-Dihydropyridines. Enzyme and Microbial Technology 2020, 132. https://doi.org/10.1016/j.enzmictec.2019.109411",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3357"
}

Simić, S., Jeremić, S., Đokić, L., Božić, N., Vujčić, Z., Lončar, N. L., Senthamaraikannan, R., Babu, R. P., Opsenica, I.,& Nikodinović-Runić, J.. (2020). Supplementary data for article: Simić, S.; Jeremic, S.; Djokic, L.; Božić, N.; Vujčić, Z.; Lončar, N.; Senthamaraikannan, R.; Babu, R.; Opsenica, I. M.; Nikodinovic-Runic, J. Development of an Efficient Biocatalytic System Based on Bacterial Laccase for the Oxidation of Selected 1,4-Dihydropyridines. Enzyme and Microbial Technology 2020, 132. https://doi.org/10.1016/j.enzmictec.2019.109411. in Enzyme and Microbial Technology.
https://hdl.handle.net/21.15107/rcub_cherry_3357

Simić S, Jeremić S, Đokić L, Božić N, Vujčić Z, Lončar NL, Senthamaraikannan R, Babu RP, Opsenica I, Nikodinović-Runić J. Supplementary data for article: Simić, S.; Jeremic, S.; Djokic, L.; Božić, N.; Vujčić, Z.; Lončar, N.; Senthamaraikannan, R.; Babu, R.; Opsenica, I. M.; Nikodinovic-Runic, J. Development of an Efficient Biocatalytic System Based on Bacterial Laccase for the Oxidation of Selected 1,4-Dihydropyridines. Enzyme and Microbial Technology 2020, 132. https://doi.org/10.1016/j.enzmictec.2019.109411. in Enzyme and Microbial Technology. 2020;.
https://hdl.handle.net/21.15107/rcub_cherry_3357 .

Simić, Stefan, Jeremić, Sanja, Đokić, Lidija, Božić, Nataša, Vujčić, Zoran, Lončar, Nikola L., Senthamaraikannan, Ramsankar, Babu, Ramesh P., Opsenica, Igor, Nikodinović-Runić, Jasmina, "Supplementary data for article: Simić, S.; Jeremic, S.; Djokic, L.; Božić, N.; Vujčić, Z.; Lončar, N.; Senthamaraikannan, R.; Babu, R.; Opsenica, I. M.; Nikodinovic-Runic, J. Development of an Efficient Biocatalytic System Based on Bacterial Laccase for the Oxidation of Selected 1,4-Dihydropyridines. Enzyme and Microbial Technology 2020, 132. https://doi.org/10.1016/j.enzmictec.2019.109411" in Enzyme and Microbial Technology (2020),
https://hdl.handle.net/21.15107/rcub_cherry_3357 .

TY  - JOUR
AU  - Simić, Stefan
AU  - Jeremić, Sanja
AU  - Đokić, Lidija
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Lončar, Nikola L.
AU  - Senthamaraikannan, Ramsankar
AU  - Babu, Ramesh P.
AU  - Opsenica, Igor
AU  - Nikodinović-Runić, Jasmina
PY  - 2020
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3356
AB  - Biocatalytic oxidations mediated by laccases are gaining importance due to their versatility and beneficial environmental effects. In this study, the oxidation of 1,4-dihydropyridines has been performed using three different types of bacterial laccase-based catalysts: purified laccase from Bacillus licheniformis ATCC 9945a (BliLacc), Escherichia coli whole cells expressing this laccase, and bacterial nanocellulose (BNC) supported BliLacc catalysts. The catalysts based on bacterial laccase were compared to the commercially available Trametes versicolor laccase (TvLacc). The oxidation product of 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylate was obtained within 7–24 h with good yields (70–99%) with all three biocatalysts. The substrate scope was examined with five additional 1,4-dihydropyridines, one of which was oxidized in high yield. Whole-cell biocatalyst was stable when stored for up to 1-month at 4 °C. In addition, evidence has been provided that multicopper oxidase CueO from the E. coli expression host contributed to the oxidation efficiency of the whole-cell biocatalyst. The immobilized whole-cell biocatalyst showed satisfactory activity and retained 37% of its original activity after three biotransformation cycles.
T2  - Enzyme and Microbial Technology
T1  - Development of an efficient biocatalytic system based on bacterial laccase for the oxidation of selected 1,4-dihydropyridines
VL  - 132
DO  - 10.1016/j.enzmictec.2019.109411
ER  - 

@article{
author = "Simić, Stefan and Jeremić, Sanja and Đokić, Lidija and Božić, Nataša and Vujčić, Zoran and Lončar, Nikola L. and Senthamaraikannan, Ramsankar and Babu, Ramesh P. and Opsenica, Igor and Nikodinović-Runić, Jasmina",
year = "2020",
abstract = "Biocatalytic oxidations mediated by laccases are gaining importance due to their versatility and beneficial environmental effects. In this study, the oxidation of 1,4-dihydropyridines has been performed using three different types of bacterial laccase-based catalysts: purified laccase from Bacillus licheniformis ATCC 9945a (BliLacc), Escherichia coli whole cells expressing this laccase, and bacterial nanocellulose (BNC) supported BliLacc catalysts. The catalysts based on bacterial laccase were compared to the commercially available Trametes versicolor laccase (TvLacc). The oxidation product of 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylate was obtained within 7–24 h with good yields (70–99%) with all three biocatalysts. The substrate scope was examined with five additional 1,4-dihydropyridines, one of which was oxidized in high yield. Whole-cell biocatalyst was stable when stored for up to 1-month at 4 °C. In addition, evidence has been provided that multicopper oxidase CueO from the E. coli expression host contributed to the oxidation efficiency of the whole-cell biocatalyst. The immobilized whole-cell biocatalyst showed satisfactory activity and retained 37% of its original activity after three biotransformation cycles.",
journal = "Enzyme and Microbial Technology",
title = "Development of an efficient biocatalytic system based on bacterial laccase for the oxidation of selected 1,4-dihydropyridines",
volume = "132",
doi = "10.1016/j.enzmictec.2019.109411"
}

Simić, S., Jeremić, S., Đokić, L., Božić, N., Vujčić, Z., Lončar, N. L., Senthamaraikannan, R., Babu, R. P., Opsenica, I.,& Nikodinović-Runić, J.. (2020). Development of an efficient biocatalytic system based on bacterial laccase for the oxidation of selected 1,4-dihydropyridines. in Enzyme and Microbial Technology, 132.
https://doi.org/10.1016/j.enzmictec.2019.109411

Simić S, Jeremić S, Đokić L, Božić N, Vujčić Z, Lončar NL, Senthamaraikannan R, Babu RP, Opsenica I, Nikodinović-Runić J. Development of an efficient biocatalytic system based on bacterial laccase for the oxidation of selected 1,4-dihydropyridines. in Enzyme and Microbial Technology. 2020;132.
doi:10.1016/j.enzmictec.2019.109411 .

Simić, Stefan, Jeremić, Sanja, Đokić, Lidija, Božić, Nataša, Vujčić, Zoran, Lončar, Nikola L., Senthamaraikannan, Ramsankar, Babu, Ramesh P., Opsenica, Igor, Nikodinović-Runić, Jasmina, "Development of an efficient biocatalytic system based on bacterial laccase for the oxidation of selected 1,4-dihydropyridines" in Enzyme and Microbial Technology, 132 (2020),
https://doi.org/10.1016/j.enzmictec.2019.109411 . .

TY  - JOUR
AU  - Božić, Nataša
AU  - Rozeboom, Henriëtte J.
AU  - Lončar, Nikola L.
AU  - Šokarda-Slavić, Marinela
AU  - Janssen, Dick B.
AU  - Vujčić, Zoran
PY  - 2020
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4261
AB  - α-Amylase from Bacillus paralicheniformis (BliAmy), belonging to GH13_5 subfamily of glycoside hydrolases, was proven to be a highly efficient raw starch digesting enzyme. The ability of some α-amylases to hydrolyze raw starch is related to the existence of surface binding sites (SBSs) for polysaccharides that can be distant from the active site. Crystallographic studies performed on BliAmy in the apo form and of enzyme bound with different oligosaccharides and oligosaccharide precursors revealed binding of these ligands to one SBS with two amino acids F257 and Y358 mainly involved in complex formation. The role of this SBS in starch binding and degradation was probed by designing enzyme variants mutated in this region (F257A and Y358A). Kinetic studies with different substrates show that starch binding through the SBS is disrupted in the mutants and that F257 and Y358 contributed cumulatively to binding and hydrolysis. Mutation of both sites (F257A/Y358A) resulted in a 5-fold lower efficacy with raw starch as substrate and at least 5.5-fold weaker binding compared to the wild type BliAmy, suggesting that the ability of BliAmy to hydrolyze raw starch with high efficiency is related to the level of its adsorption onto starch granules.
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T1  - Characterization of the starch surface binding site on Bacillus paralicheniformis α-amylase
VL  - 165
IS  - A
SP  - 1529
EP  - 1539
DO  - 10.1016/j.ijbiomac.2020.10.025
ER  - 

@article{
author = "Božić, Nataša and Rozeboom, Henriëtte J. and Lončar, Nikola L. and Šokarda-Slavić, Marinela and Janssen, Dick B. and Vujčić, Zoran",
year = "2020",
abstract = "α-Amylase from Bacillus paralicheniformis (BliAmy), belonging to GH13_5 subfamily of glycoside hydrolases, was proven to be a highly efficient raw starch digesting enzyme. The ability of some α-amylases to hydrolyze raw starch is related to the existence of surface binding sites (SBSs) for polysaccharides that can be distant from the active site. Crystallographic studies performed on BliAmy in the apo form and of enzyme bound with different oligosaccharides and oligosaccharide precursors revealed binding of these ligands to one SBS with two amino acids F257 and Y358 mainly involved in complex formation. The role of this SBS in starch binding and degradation was probed by designing enzyme variants mutated in this region (F257A and Y358A). Kinetic studies with different substrates show that starch binding through the SBS is disrupted in the mutants and that F257 and Y358 contributed cumulatively to binding and hydrolysis. Mutation of both sites (F257A/Y358A) resulted in a 5-fold lower efficacy with raw starch as substrate and at least 5.5-fold weaker binding compared to the wild type BliAmy, suggesting that the ability of BliAmy to hydrolyze raw starch with high efficiency is related to the level of its adsorption onto starch granules.",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules",
title = "Characterization of the starch surface binding site on Bacillus paralicheniformis α-amylase",
volume = "165",
number = "A",
pages = "1529-1539",
doi = "10.1016/j.ijbiomac.2020.10.025"
}

Božić, N., Rozeboom, H. J., Lončar, N. L., Šokarda-Slavić, M., Janssen, D. B.,& Vujčić, Z.. (2020). Characterization of the starch surface binding site on Bacillus paralicheniformis α-amylase. in International Journal of Biological Macromolecules
Elsevier., 165(A), 1529-1539.
https://doi.org/10.1016/j.ijbiomac.2020.10.025

Božić N, Rozeboom HJ, Lončar NL, Šokarda-Slavić M, Janssen DB, Vujčić Z. Characterization of the starch surface binding site on Bacillus paralicheniformis α-amylase. in International Journal of Biological Macromolecules. 2020;165(A):1529-1539.
doi:10.1016/j.ijbiomac.2020.10.025 .

Božić, Nataša, Rozeboom, Henriëtte J., Lončar, Nikola L., Šokarda-Slavić, Marinela, Janssen, Dick B., Vujčić, Zoran, "Characterization of the starch surface binding site on Bacillus paralicheniformis α-amylase" in International Journal of Biological Macromolecules, 165, no. A (2020):1529-1539,
https://doi.org/10.1016/j.ijbiomac.2020.10.025 . .

TY  - JOUR
AU  - Božić, Nataša
AU  - Rozeboom, Henriëtte J.
AU  - Lončar, Nikola L.
AU  - Šokarda-Slavić, Marinela
AU  - Janssen, Dick B.
AU  - Vujčić, Zoran
PY  - 2020
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4262
AB  - α-Amylase from Bacillus paralicheniformis (BliAmy), belonging to GH13_5 subfamily of glycoside hydrolases, was proven to be a highly efficient raw starch digesting enzyme. The ability of some α-amylases to hydrolyze raw starch is related to the existence of surface binding sites (SBSs) for polysaccharides that can be distant from the active site. Crystallographic studies performed on BliAmy in the apo form and of enzyme bound with different oligosaccharides and oligosaccharide precursors revealed binding of these ligands to one SBS with two amino acids F257 and Y358 mainly involved in complex formation. The role of this SBS in starch binding and degradation was probed by designing enzyme variants mutated in this region (F257A and Y358A). Kinetic studies with different substrates show that starch binding through the SBS is disrupted in the mutants and that F257 and Y358 contributed cumulatively to binding and hydrolysis. Mutation of both sites (F257A/Y358A) resulted in a 5-fold lower efficacy with raw starch as substrate and at least 5.5-fold weaker binding compared to the wild type BliAmy, suggesting that the ability of BliAmy to hydrolyze raw starch with high efficiency is related to the level of its adsorption onto starch granules.
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T1  - Characterization of the starch surface binding site on Bacillus paralicheniformis α-amylase
VL  - 165
IS  - A
SP  - 1529
EP  - 1539
DO  - 10.1016/j.ijbiomac.2020.10.025
ER  - 

@article{
author = "Božić, Nataša and Rozeboom, Henriëtte J. and Lončar, Nikola L. and Šokarda-Slavić, Marinela and Janssen, Dick B. and Vujčić, Zoran",
year = "2020",
abstract = "α-Amylase from Bacillus paralicheniformis (BliAmy), belonging to GH13_5 subfamily of glycoside hydrolases, was proven to be a highly efficient raw starch digesting enzyme. The ability of some α-amylases to hydrolyze raw starch is related to the existence of surface binding sites (SBSs) for polysaccharides that can be distant from the active site. Crystallographic studies performed on BliAmy in the apo form and of enzyme bound with different oligosaccharides and oligosaccharide precursors revealed binding of these ligands to one SBS with two amino acids F257 and Y358 mainly involved in complex formation. The role of this SBS in starch binding and degradation was probed by designing enzyme variants mutated in this region (F257A and Y358A). Kinetic studies with different substrates show that starch binding through the SBS is disrupted in the mutants and that F257 and Y358 contributed cumulatively to binding and hydrolysis. Mutation of both sites (F257A/Y358A) resulted in a 5-fold lower efficacy with raw starch as substrate and at least 5.5-fold weaker binding compared to the wild type BliAmy, suggesting that the ability of BliAmy to hydrolyze raw starch with high efficiency is related to the level of its adsorption onto starch granules.",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules",
title = "Characterization of the starch surface binding site on Bacillus paralicheniformis α-amylase",
volume = "165",
number = "A",
pages = "1529-1539",
doi = "10.1016/j.ijbiomac.2020.10.025"
}

Božić, N., Rozeboom, H. J., Lončar, N. L., Šokarda-Slavić, M., Janssen, D. B.,& Vujčić, Z.. (2020). Characterization of the starch surface binding site on Bacillus paralicheniformis α-amylase. in International Journal of Biological Macromolecules
Elsevier., 165(A), 1529-1539.
https://doi.org/10.1016/j.ijbiomac.2020.10.025

Božić N, Rozeboom HJ, Lončar NL, Šokarda-Slavić M, Janssen DB, Vujčić Z. Characterization of the starch surface binding site on Bacillus paralicheniformis α-amylase. in International Journal of Biological Macromolecules. 2020;165(A):1529-1539.
doi:10.1016/j.ijbiomac.2020.10.025 .

Božić, Nataša, Rozeboom, Henriëtte J., Lončar, Nikola L., Šokarda-Slavić, Marinela, Janssen, Dick B., Vujčić, Zoran, "Characterization of the starch surface binding site on Bacillus paralicheniformis α-amylase" in International Journal of Biological Macromolecules, 165, no. A (2020):1529-1539,
https://doi.org/10.1016/j.ijbiomac.2020.10.025 . .

TY  - JOUR
AU  - Lončar, Nikola L.
AU  - Drašković, Natalija
AU  - Božić, Nataša
AU  - Romero, Elvira
AU  - Simić, Stefan
AU  - Opsenica, Igor
AU  - Vujčić, Zoran
AU  - Fraaije, Marco W.
PY  - 2019
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3709
AB  - The consumption of dyes is increasing worldwide in line with the increase of population and demand for clothes and other colored products. However, the efficiency of dyeing processes is still poor and results in large amounts of colored effluents. It is desired to develop a portfolio of enzymes which can be used for the treatment of colored wastewaters. Herein, we used genome sequence information to discover a dye-decolorizing peroxidase (DyP) from Pseudomonas fluorescens Pf-01. Two genes putatively encoding for DyPs were identified in the respective genome and cloned for expression in Escherichia coli, of which one (Pf DyP B2) could be overexpressed as a soluble protein. Pf DyP B2 shows some typical features known for DyPs which includes the ability to convert dyes at the expense of hydrogen peroxide. Interestingly, t-butyl hydroperoxide could be used as an alternative substrate to hydrogen peroxide. Immobilization of Pf DyP B2 in calcium-alginate beads resulted in a significant increase in stability: Pf DyP B2 retains 80% of its initial activity after 2 h incubation at 50◦ C, while the soluble enzyme is inactivated within minutes. Pf DyP B2 was also tested with aniline and ethyl diazoacetate as substrates. Based on GC-MS analyses, 30% conversion of the starting material was achieved after 65 h at 30◦ C. Importantly, this is the first report of a DyP-catalyzed insertion of a carbene into an N-H bond.
PB  - MDPI
T2  - Catalysts
T1  - Expression and characterization of a dye-decolorizing peroxidase from pseudomonas fluorescens Pf0-1
VL  - 9
IS  - 5
DO  - 10.3390/catal9050463
ER  - 

@article{
author = "Lončar, Nikola L. and Drašković, Natalija and Božić, Nataša and Romero, Elvira and Simić, Stefan and Opsenica, Igor and Vujčić, Zoran and Fraaije, Marco W.",
year = "2019",
abstract = "The consumption of dyes is increasing worldwide in line with the increase of population and demand for clothes and other colored products. However, the efficiency of dyeing processes is still poor and results in large amounts of colored effluents. It is desired to develop a portfolio of enzymes which can be used for the treatment of colored wastewaters. Herein, we used genome sequence information to discover a dye-decolorizing peroxidase (DyP) from Pseudomonas fluorescens Pf-01. Two genes putatively encoding for DyPs were identified in the respective genome and cloned for expression in Escherichia coli, of which one (Pf DyP B2) could be overexpressed as a soluble protein. Pf DyP B2 shows some typical features known for DyPs which includes the ability to convert dyes at the expense of hydrogen peroxide. Interestingly, t-butyl hydroperoxide could be used as an alternative substrate to hydrogen peroxide. Immobilization of Pf DyP B2 in calcium-alginate beads resulted in a significant increase in stability: Pf DyP B2 retains 80% of its initial activity after 2 h incubation at 50◦ C, while the soluble enzyme is inactivated within minutes. Pf DyP B2 was also tested with aniline and ethyl diazoacetate as substrates. Based on GC-MS analyses, 30% conversion of the starting material was achieved after 65 h at 30◦ C. Importantly, this is the first report of a DyP-catalyzed insertion of a carbene into an N-H bond.",
publisher = "MDPI",
journal = "Catalysts",
title = "Expression and characterization of a dye-decolorizing peroxidase from pseudomonas fluorescens Pf0-1",
volume = "9",
number = "5",
doi = "10.3390/catal9050463"
}

Lončar, N. L., Drašković, N., Božić, N., Romero, E., Simić, S., Opsenica, I., Vujčić, Z.,& Fraaije, M. W.. (2019). Expression and characterization of a dye-decolorizing peroxidase from pseudomonas fluorescens Pf0-1. in Catalysts
MDPI., 9(5).
https://doi.org/10.3390/catal9050463

Lončar NL, Drašković N, Božić N, Romero E, Simić S, Opsenica I, Vujčić Z, Fraaije MW. Expression and characterization of a dye-decolorizing peroxidase from pseudomonas fluorescens Pf0-1. in Catalysts. 2019;9(5).
doi:10.3390/catal9050463 .

Lončar, Nikola L., Drašković, Natalija, Božić, Nataša, Romero, Elvira, Simić, Stefan, Opsenica, Igor, Vujčić, Zoran, Fraaije, Marco W., "Expression and characterization of a dye-decolorizing peroxidase from pseudomonas fluorescens Pf0-1" in Catalysts, 9, no. 5 (2019),
https://doi.org/10.3390/catal9050463 . .

TY  - DATA
AU  - Lončar, Nikola L.
AU  - Drašković, Natalija
AU  - Božić, Nataša
AU  - Romero, Elvira
AU  - Simić, Stefan
AU  - Opsenica, Igor
AU  - Vujčić, Zoran
AU  - Fraaije, Marco W.
PY  - 2019
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3710
PB  - MDPI
T2  - Catalysts
T1  - Supplementary data for the article: Lončar, N.; Drašković, N.; Božić, N.; Romero, E.; Simić, S.; Opsenica, I.; Vujčić, Z.; Fraaije, M. W. Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1. Catalysts 2019, 9 (5). https://doi.org/10.3390/catal9050463
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3710
ER  - 

@misc{
author = "Lončar, Nikola L. and Drašković, Natalija and Božić, Nataša and Romero, Elvira and Simić, Stefan and Opsenica, Igor and Vujčić, Zoran and Fraaije, Marco W.",
year = "2019",
publisher = "MDPI",
journal = "Catalysts",
title = "Supplementary data for the article: Lončar, N.; Drašković, N.; Božić, N.; Romero, E.; Simić, S.; Opsenica, I.; Vujčić, Z.; Fraaije, M. W. Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1. Catalysts 2019, 9 (5). https://doi.org/10.3390/catal9050463",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3710"
}

Lončar, N. L., Drašković, N., Božić, N., Romero, E., Simić, S., Opsenica, I., Vujčić, Z.,& Fraaije, M. W.. (2019). Supplementary data for the article: Lončar, N.; Drašković, N.; Božić, N.; Romero, E.; Simić, S.; Opsenica, I.; Vujčić, Z.; Fraaije, M. W. Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1. Catalysts 2019, 9 (5). https://doi.org/10.3390/catal9050463. in Catalysts
MDPI..
https://hdl.handle.net/21.15107/rcub_cherry_3710

Lončar NL, Drašković N, Božić N, Romero E, Simić S, Opsenica I, Vujčić Z, Fraaije MW. Supplementary data for the article: Lončar, N.; Drašković, N.; Božić, N.; Romero, E.; Simić, S.; Opsenica, I.; Vujčić, Z.; Fraaije, M. W. Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1. Catalysts 2019, 9 (5). https://doi.org/10.3390/catal9050463. in Catalysts. 2019;.
https://hdl.handle.net/21.15107/rcub_cherry_3710 .

Lončar, Nikola L., Drašković, Natalija, Božić, Nataša, Romero, Elvira, Simić, Stefan, Opsenica, Igor, Vujčić, Zoran, Fraaije, Marco W., "Supplementary data for the article: Lončar, N.; Drašković, N.; Božić, N.; Romero, E.; Simić, S.; Opsenica, I.; Vujčić, Z.; Fraaije, M. W. Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1. Catalysts 2019, 9 (5). https://doi.org/10.3390/catal9050463" in Catalysts (2019),
https://hdl.handle.net/21.15107/rcub_cherry_3710 .

TY  - JOUR
AU  - Šokarda-Slavić, Marinela
AU  - Pešić, Milja
AU  - Vujčić, Zoran
AU  - Božić, Nataša
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2061
AB  - alpha-Amylase from Bacillus licheniformis ATCC 9945a (BliAmy) was proven to be very efficient in hydrolysis of granular starch below the temperature of gelatinization. By applying two-stage feeding strategy to achieve high-cell-density cultivation of Escherichia coli and extracellular production of BliAmy, total of 250.5 U/mL (i.e. 0.7 g/L) of enzyme was obtained. Thermostability of amylase was exploited to simplify purification. The hydrolysis of concentrated raw starch was optimized using response surface methodology. Regardless of raw starch concentration tested (20, 25, 30 %), BliAmy was very effective, achieving the final hydrolysis degree of 91 % for the hydrolysis of 30 % starch suspension after 24 h. The major A-type crystalline structure and amorphous domains of the starch granule were degraded at the same rates, while amylose-lipid complexes were not degraded. BliAmy presents interesting performances on highly concentrated solid starch and could be of value for starch-consuming industries while response surface methodology (RSM) could be efficiently applied for the optimization of the hydrolysis.
PB  - Springer, New York
T2  - Applied Microbiology and Biotechnology
T1  - Overcoming hydrolysis of raw corn starch under industrial conditions with Bacillus licheniformis ATCC 9945a alpha-amylase
VL  - 100
IS  - 6
SP  - 2709
EP  - 2719
DO  - 10.1007/s00253-015-7101-4
ER  - 

@article{
author = "Šokarda-Slavić, Marinela and Pešić, Milja and Vujčić, Zoran and Božić, Nataša",
year = "2016",
abstract = "alpha-Amylase from Bacillus licheniformis ATCC 9945a (BliAmy) was proven to be very efficient in hydrolysis of granular starch below the temperature of gelatinization. By applying two-stage feeding strategy to achieve high-cell-density cultivation of Escherichia coli and extracellular production of BliAmy, total of 250.5 U/mL (i.e. 0.7 g/L) of enzyme was obtained. Thermostability of amylase was exploited to simplify purification. The hydrolysis of concentrated raw starch was optimized using response surface methodology. Regardless of raw starch concentration tested (20, 25, 30 %), BliAmy was very effective, achieving the final hydrolysis degree of 91 % for the hydrolysis of 30 % starch suspension after 24 h. The major A-type crystalline structure and amorphous domains of the starch granule were degraded at the same rates, while amylose-lipid complexes were not degraded. BliAmy presents interesting performances on highly concentrated solid starch and could be of value for starch-consuming industries while response surface methodology (RSM) could be efficiently applied for the optimization of the hydrolysis.",
publisher = "Springer, New York",
journal = "Applied Microbiology and Biotechnology",
title = "Overcoming hydrolysis of raw corn starch under industrial conditions with Bacillus licheniformis ATCC 9945a alpha-amylase",
volume = "100",
number = "6",
pages = "2709-2719",
doi = "10.1007/s00253-015-7101-4"
}

Šokarda-Slavić, M., Pešić, M., Vujčić, Z.,& Božić, N.. (2016). Overcoming hydrolysis of raw corn starch under industrial conditions with Bacillus licheniformis ATCC 9945a alpha-amylase. in Applied Microbiology and Biotechnology
Springer, New York., 100(6), 2709-2719.
https://doi.org/10.1007/s00253-015-7101-4

Šokarda-Slavić M, Pešić M, Vujčić Z, Božić N. Overcoming hydrolysis of raw corn starch under industrial conditions with Bacillus licheniformis ATCC 9945a alpha-amylase. in Applied Microbiology and Biotechnology. 2016;100(6):2709-2719.
doi:10.1007/s00253-015-7101-4 .

Šokarda-Slavić, Marinela, Pešić, Milja, Vujčić, Zoran, Božić, Nataša, "Overcoming hydrolysis of raw corn starch under industrial conditions with Bacillus licheniformis ATCC 9945a alpha-amylase" in Applied Microbiology and Biotechnology, 100, no. 6 (2016):2709-2719,
https://doi.org/10.1007/s00253-015-7101-4 . .

TY  - JOUR
AU  - Lončar, Nikola L.
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2375
AB  - Bacterial laccases have proven advantages over fungal and plant counterparts in terms of wider pH optimum, higher stability and broader biocatalytic scope. In this work, Bacillus licheniformis ATCC 9945a laccase is produced heterologously in Escherichia coli. Produced laccase exhibits remarkably high temperature optimum at 90 degrees C and possess significant thermostability and resistance to inactivation by organic solvents. Laccase has an apparent melting temperature of 79 degrees C at pH 7.0 and above 70 degrees C in range of pH 5.0-8.0, while having half-life of 50 min at 70 degrees C. Presence of 10% organic solvents such as acetonitrile, dimethylformamide, dimethylsulfoxide or methanol reduces melting temperature to 45-52 degrees C but activity remains practically unimpaired. With 50% of acetonitrile and methanol laccase retained similar to 40% of initial activity. EDTA and 300 mM sodium -chloride have positive effect on activity. Enzyme is active on syringaldazine, ABTS, phenols, amines, naphthol, lignin and lignin model compounds and mediates C-C bond formation via oxidative coupling after one electron oxidation of phenolic group. Successful polymerization of 2 -naphthol was achieved with 77% conversion of 250 mg/L 2-naphtol in only 15 min which may further expand substrate scope of this enzyme towards polymer production and/or xenobiotics removal for environmental applications. (C) 2016 Elsevier B.V. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - Journal of Molecular Catalysis. B: Enzymatic
T1  - Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a
VL  - 134
SP  - 390
EP  - 395
DO  - 10.1016/j.molcatb.2016.06.005
ER  - 

@article{
author = "Lončar, Nikola L. and Božić, Nataša and Vujčić, Zoran",
year = "2016",
abstract = "Bacterial laccases have proven advantages over fungal and plant counterparts in terms of wider pH optimum, higher stability and broader biocatalytic scope. In this work, Bacillus licheniformis ATCC 9945a laccase is produced heterologously in Escherichia coli. Produced laccase exhibits remarkably high temperature optimum at 90 degrees C and possess significant thermostability and resistance to inactivation by organic solvents. Laccase has an apparent melting temperature of 79 degrees C at pH 7.0 and above 70 degrees C in range of pH 5.0-8.0, while having half-life of 50 min at 70 degrees C. Presence of 10% organic solvents such as acetonitrile, dimethylformamide, dimethylsulfoxide or methanol reduces melting temperature to 45-52 degrees C but activity remains practically unimpaired. With 50% of acetonitrile and methanol laccase retained similar to 40% of initial activity. EDTA and 300 mM sodium -chloride have positive effect on activity. Enzyme is active on syringaldazine, ABTS, phenols, amines, naphthol, lignin and lignin model compounds and mediates C-C bond formation via oxidative coupling after one electron oxidation of phenolic group. Successful polymerization of 2 -naphthol was achieved with 77% conversion of 250 mg/L 2-naphtol in only 15 min which may further expand substrate scope of this enzyme towards polymer production and/or xenobiotics removal for environmental applications. (C) 2016 Elsevier B.V. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Journal of Molecular Catalysis. B: Enzymatic",
title = "Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a",
volume = "134",
pages = "390-395",
doi = "10.1016/j.molcatb.2016.06.005"
}

Lončar, N. L., Božić, N.,& Vujčić, Z.. (2016). Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a. in Journal of Molecular Catalysis. B: Enzymatic
Elsevier Science Bv, Amsterdam., 134, 390-395.
https://doi.org/10.1016/j.molcatb.2016.06.005

Lončar NL, Božić N, Vujčić Z. Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a. in Journal of Molecular Catalysis. B: Enzymatic. 2016;134:390-395.
doi:10.1016/j.molcatb.2016.06.005 .

Lončar, Nikola L., Božić, Nataša, Vujčić, Zoran, "Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a" in Journal of Molecular Catalysis. B: Enzymatic, 134 (2016):390-395,
https://doi.org/10.1016/j.molcatb.2016.06.005 . .

TY  - JOUR
AU  - Lončar, Nikola L.
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3517
AB  - Bacterial laccases have proven advantages over fungal and plant counterparts in terms of wider pH optimum, higher stability and broader biocatalytic scope. In this work, Bacillus licheniformis ATCC 9945a laccase is produced heterologously in Escherichia coli. Produced laccase exhibits remarkably high temperature optimum at 90 degrees C and possess significant thermostability and resistance to inactivation by organic solvents. Laccase has an apparent melting temperature of 79 degrees C at pH 7.0 and above 70 degrees C in range of pH 5.0-8.0, while having half-life of 50 min at 70 degrees C. Presence of 10% organic solvents such as acetonitrile, dimethylformamide, dimethylsulfoxide or methanol reduces melting temperature to 45-52 degrees C but activity remains practically unimpaired. With 50% of acetonitrile and methanol laccase retained similar to 40% of initial activity. EDTA and 300 mM sodium -chloride have positive effect on activity. Enzyme is active on syringaldazine, ABTS, phenols, amines, naphthol, lignin and lignin model compounds and mediates C-C bond formation via oxidative coupling after one electron oxidation of phenolic group. Successful polymerization of 2 -naphthol was achieved with 77% conversion of 250 mg/L 2-naphtol in only 15 min which may further expand substrate scope of this enzyme towards polymer production and/or xenobiotics removal for environmental applications. (C) 2016 Elsevier B.V. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - Journal of Molecular Catalysis. B: Enzymatic
T1  - Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a
VL  - 134
SP  - 390
EP  - 395
DO  - 10.1016/j.molcatb.2016.06.005
ER  - 

@article{
author = "Lončar, Nikola L. and Božić, Nataša and Vujčić, Zoran",
year = "2016",
abstract = "Bacterial laccases have proven advantages over fungal and plant counterparts in terms of wider pH optimum, higher stability and broader biocatalytic scope. In this work, Bacillus licheniformis ATCC 9945a laccase is produced heterologously in Escherichia coli. Produced laccase exhibits remarkably high temperature optimum at 90 degrees C and possess significant thermostability and resistance to inactivation by organic solvents. Laccase has an apparent melting temperature of 79 degrees C at pH 7.0 and above 70 degrees C in range of pH 5.0-8.0, while having half-life of 50 min at 70 degrees C. Presence of 10% organic solvents such as acetonitrile, dimethylformamide, dimethylsulfoxide or methanol reduces melting temperature to 45-52 degrees C but activity remains practically unimpaired. With 50% of acetonitrile and methanol laccase retained similar to 40% of initial activity. EDTA and 300 mM sodium -chloride have positive effect on activity. Enzyme is active on syringaldazine, ABTS, phenols, amines, naphthol, lignin and lignin model compounds and mediates C-C bond formation via oxidative coupling after one electron oxidation of phenolic group. Successful polymerization of 2 -naphthol was achieved with 77% conversion of 250 mg/L 2-naphtol in only 15 min which may further expand substrate scope of this enzyme towards polymer production and/or xenobiotics removal for environmental applications. (C) 2016 Elsevier B.V. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Journal of Molecular Catalysis. B: Enzymatic",
title = "Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a",
volume = "134",
pages = "390-395",
doi = "10.1016/j.molcatb.2016.06.005"
}

Lončar, N. L., Božić, N.,& Vujčić, Z.. (2016). Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a. in Journal of Molecular Catalysis. B: Enzymatic
Elsevier Science Bv, Amsterdam., 134, 390-395.
https://doi.org/10.1016/j.molcatb.2016.06.005

Lončar NL, Božić N, Vujčić Z. Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a. in Journal of Molecular Catalysis. B: Enzymatic. 2016;134:390-395.
doi:10.1016/j.molcatb.2016.06.005 .

Lončar, Nikola L., Božić, Nataša, Vujčić, Zoran, "Expression and characterization of a thermostable organic solvent-tolerant laccase from Bacillus licheniformis ATCC 9945a" in Journal of Molecular Catalysis. B: Enzymatic, 134 (2016):390-395,
https://doi.org/10.1016/j.molcatb.2016.06.005 . .

TY  - DATA
AU  - Lončar, Nikola L.
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3518
PB  - Elsevier Science Bv, Amsterdam
T2  - Journal of Molecular Catalysis. B: Enzymatic
T1  - Supplementary data for the article: Loncar, N.; Bozic, N.; Vujcic, Z. Expression and Characterization of a Thermostable Organic Solvent-Tolerant Laccase from Bacillus Licheniformis ATCC 9945a. J. Mol. Catal. B-Enzym. 2016, 134, 390–395. https://doi.org/10.1016/j.molcatb.2016.06.005
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3518
ER  - 

@misc{
author = "Lončar, Nikola L. and Božić, Nataša and Vujčić, Zoran",
year = "2016",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Journal of Molecular Catalysis. B: Enzymatic",
title = "Supplementary data for the article: Loncar, N.; Bozic, N.; Vujcic, Z. Expression and Characterization of a Thermostable Organic Solvent-Tolerant Laccase from Bacillus Licheniformis ATCC 9945a. J. Mol. Catal. B-Enzym. 2016, 134, 390–395. https://doi.org/10.1016/j.molcatb.2016.06.005",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3518"
}

Lončar, N. L., Božić, N.,& Vujčić, Z.. (2016). Supplementary data for the article: Loncar, N.; Bozic, N.; Vujcic, Z. Expression and Characterization of a Thermostable Organic Solvent-Tolerant Laccase from Bacillus Licheniformis ATCC 9945a. J. Mol. Catal. B-Enzym. 2016, 134, 390–395. https://doi.org/10.1016/j.molcatb.2016.06.005. in Journal of Molecular Catalysis. B: Enzymatic
Elsevier Science Bv, Amsterdam..
https://hdl.handle.net/21.15107/rcub_cherry_3518

Lončar NL, Božić N, Vujčić Z. Supplementary data for the article: Loncar, N.; Bozic, N.; Vujcic, Z. Expression and Characterization of a Thermostable Organic Solvent-Tolerant Laccase from Bacillus Licheniformis ATCC 9945a. J. Mol. Catal. B-Enzym. 2016, 134, 390–395. https://doi.org/10.1016/j.molcatb.2016.06.005. in Journal of Molecular Catalysis. B: Enzymatic. 2016;.
https://hdl.handle.net/21.15107/rcub_cherry_3518 .

Lončar, Nikola L., Božić, Nataša, Vujčić, Zoran, "Supplementary data for the article: Loncar, N.; Bozic, N.; Vujcic, Z. Expression and Characterization of a Thermostable Organic Solvent-Tolerant Laccase from Bacillus Licheniformis ATCC 9945a. J. Mol. Catal. B-Enzym. 2016, 134, 390–395. https://doi.org/10.1016/j.molcatb.2016.06.005" in Journal of Molecular Catalysis. B: Enzymatic (2016),
https://hdl.handle.net/21.15107/rcub_cherry_3518 .

TY  - DATA
AU  - Šokarda-Slavić, Marinela
AU  - Pešić, Milja
AU  - Vujčić, Zoran
AU  - Božić, Nataša
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3588
PB  - Springer, New York
T2  - Applied Microbiology and Biotechnology
T1  - Supplementary material for the article: Šokarda Slavić, M.; Pešić, M.; Vujčić, Z.; Božić, N. Overcoming Hydrolysis of Raw Corn  Starch under Industrial Conditions with Bacillus Licheniformis ATCC 9945a α-Amylase. Applied Microbiology and Biotechnology 2016, 100 (6), 2709–2719. https://doi.org/10.1007/s00253-015-7101-4
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3588
ER  - 

@misc{
author = "Šokarda-Slavić, Marinela and Pešić, Milja and Vujčić, Zoran and Božić, Nataša",
year = "2016",
publisher = "Springer, New York",
journal = "Applied Microbiology and Biotechnology",
title = "Supplementary material for the article: Šokarda Slavić, M.; Pešić, M.; Vujčić, Z.; Božić, N. Overcoming Hydrolysis of Raw Corn  Starch under Industrial Conditions with Bacillus Licheniformis ATCC 9945a α-Amylase. Applied Microbiology and Biotechnology 2016, 100 (6), 2709–2719. https://doi.org/10.1007/s00253-015-7101-4",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3588"
}

Šokarda-Slavić, M., Pešić, M., Vujčić, Z.,& Božić, N.. (2016). Supplementary material for the article: Šokarda Slavić, M.; Pešić, M.; Vujčić, Z.; Božić, N. Overcoming Hydrolysis of Raw Corn  Starch under Industrial Conditions with Bacillus Licheniformis ATCC 9945a α-Amylase. Applied Microbiology and Biotechnology 2016, 100 (6), 2709–2719. https://doi.org/10.1007/s00253-015-7101-4. in Applied Microbiology and Biotechnology
Springer, New York..
https://hdl.handle.net/21.15107/rcub_cherry_3588

Šokarda-Slavić M, Pešić M, Vujčić Z, Božić N. Supplementary material for the article: Šokarda Slavić, M.; Pešić, M.; Vujčić, Z.; Božić, N. Overcoming Hydrolysis of Raw Corn  Starch under Industrial Conditions with Bacillus Licheniformis ATCC 9945a α-Amylase. Applied Microbiology and Biotechnology 2016, 100 (6), 2709–2719. https://doi.org/10.1007/s00253-015-7101-4. in Applied Microbiology and Biotechnology. 2016;.
https://hdl.handle.net/21.15107/rcub_cherry_3588 .

Šokarda-Slavić, Marinela, Pešić, Milja, Vujčić, Zoran, Božić, Nataša, "Supplementary material for the article: Šokarda Slavić, M.; Pešić, M.; Vujčić, Z.; Božić, N. Overcoming Hydrolysis of Raw Corn  Starch under Industrial Conditions with Bacillus Licheniformis ATCC 9945a α-Amylase. Applied Microbiology and Biotechnology 2016, 100 (6), 2709–2719. https://doi.org/10.1007/s00253-015-7101-4" in Applied Microbiology and Biotechnology (2016),
https://hdl.handle.net/21.15107/rcub_cherry_3588 .

TY  - JOUR
AU  - Vujčić, Zoran
AU  - Janović, Barbara
AU  - Lončar, Nikola L.
AU  - Margetić, Aleksandra
AU  - Božić, Nataša
AU  - Dojnov, Biljana
AU  - Vujčić, Miroslava
PY  - 2015
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1644
AB  - Horseradish peroxidase (HRP) is enzyme first described more than 200 years ago and yet there are still some aspects of this potent enzyme to be tackled. Researchers were focused on most abundant isoenzyme HRP CIA while remaining, particularly anionic isoenzymes were discarded in purification process. This work describes exploitation of those isoenzymes for removal of recalcitrant pollutants such as reactive dyes. Results demonstrated that not only these enzymes can decolorize dyes but also in some cases anionic forms are more efficient than commercially produced cationic HRP form. Enzyme concentration of 0.14 U ml(-1) was found to provide maximum dye removal at optimized reaction conditions with dye concentration of 30 mg I-1. Majority of dyes tested were successfully decolorized at pH 5 or 7 while some dyes like Orange 2 and Reactive black 5 are decolorized most efficiently at pH 9. Anionic isoenzymes act by disrupting chromophore of Reactive black 5 while cationic HRP oxidize dye but leaves chromophore present. (C) 2014 Elsevier Ltd. All rights reserved.
PB  - Elsevier Sci Ltd, Oxford
T2  - International Biodeterioration and Biodegradation
T1  - Exploitation of neglected horseradish peroxidase izoenzymes for dye decolorization
VL  - 97
SP  - 124
EP  - 127
DO  - 10.1016/j.ibiod.2014.10.007
ER  - 

@article{
author = "Vujčić, Zoran and Janović, Barbara and Lončar, Nikola L. and Margetić, Aleksandra and Božić, Nataša and Dojnov, Biljana and Vujčić, Miroslava",
year = "2015",
abstract = "Horseradish peroxidase (HRP) is enzyme first described more than 200 years ago and yet there are still some aspects of this potent enzyme to be tackled. Researchers were focused on most abundant isoenzyme HRP CIA while remaining, particularly anionic isoenzymes were discarded in purification process. This work describes exploitation of those isoenzymes for removal of recalcitrant pollutants such as reactive dyes. Results demonstrated that not only these enzymes can decolorize dyes but also in some cases anionic forms are more efficient than commercially produced cationic HRP form. Enzyme concentration of 0.14 U ml(-1) was found to provide maximum dye removal at optimized reaction conditions with dye concentration of 30 mg I-1. Majority of dyes tested were successfully decolorized at pH 5 or 7 while some dyes like Orange 2 and Reactive black 5 are decolorized most efficiently at pH 9. Anionic isoenzymes act by disrupting chromophore of Reactive black 5 while cationic HRP oxidize dye but leaves chromophore present. (C) 2014 Elsevier Ltd. All rights reserved.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "International Biodeterioration and Biodegradation",
title = "Exploitation of neglected horseradish peroxidase izoenzymes for dye decolorization",
volume = "97",
pages = "124-127",
doi = "10.1016/j.ibiod.2014.10.007"
}

Vujčić, Z., Janović, B., Lončar, N. L., Margetić, A., Božić, N., Dojnov, B.,& Vujčić, M.. (2015). Exploitation of neglected horseradish peroxidase izoenzymes for dye decolorization. in International Biodeterioration and Biodegradation
Elsevier Sci Ltd, Oxford., 97, 124-127.
https://doi.org/10.1016/j.ibiod.2014.10.007

Vujčić Z, Janović B, Lončar NL, Margetić A, Božić N, Dojnov B, Vujčić M. Exploitation of neglected horseradish peroxidase izoenzymes for dye decolorization. in International Biodeterioration and Biodegradation. 2015;97:124-127.
doi:10.1016/j.ibiod.2014.10.007 .

Vujčić, Zoran, Janović, Barbara, Lončar, Nikola L., Margetić, Aleksandra, Božić, Nataša, Dojnov, Biljana, Vujčić, Miroslava, "Exploitation of neglected horseradish peroxidase izoenzymes for dye decolorization" in International Biodeterioration and Biodegradation, 97 (2015):124-127,
https://doi.org/10.1016/j.ibiod.2014.10.007 . .

TY  - JOUR
AU  - Lončar, Nikola L.
AU  - Šokarda-Slavić, Marinela
AU  - Vujčić, Zoran
AU  - Božić, Nataša
PY  - 2015
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1985
AB  - Bacillus licheniformis 9945a alpha-amylase is known as a potent enzyme for raw starch hydrolysis. In this paper, a mixed mode Nuvia cPrime (TM) resin is examined with the aim to improve the downstream processing of raw starch digesting amylases and exploit the hydrophobic patches on their surface. This resin combines hydrophobic interactions with cation exchange groups and as such the presence of salt facilitates hydrophobic interactions while the ion-exchange groups enable proper selectivity. alpha-Amylase was produced using an optimized fed-batch approach in a defined media and significant overexpression of 1.2 g L-1 was achieved. This single step procedure enables simultaneous concentration, pigment removal as well as purification of amylase with yields of 96% directly from the fermentation broth.
PB  - Nature Publishing Group, London
T2  - Scientific Reports
T1  - Mixed-mode resins: taking shortcut in downstream processing of raw-starch digesting alpha-amylases
VL  - 5
DO  - 10.1038/srep15772
ER  - 

@article{
author = "Lončar, Nikola L. and Šokarda-Slavić, Marinela and Vujčić, Zoran and Božić, Nataša",
year = "2015",
abstract = "Bacillus licheniformis 9945a alpha-amylase is known as a potent enzyme for raw starch hydrolysis. In this paper, a mixed mode Nuvia cPrime (TM) resin is examined with the aim to improve the downstream processing of raw starch digesting amylases and exploit the hydrophobic patches on their surface. This resin combines hydrophobic interactions with cation exchange groups and as such the presence of salt facilitates hydrophobic interactions while the ion-exchange groups enable proper selectivity. alpha-Amylase was produced using an optimized fed-batch approach in a defined media and significant overexpression of 1.2 g L-1 was achieved. This single step procedure enables simultaneous concentration, pigment removal as well as purification of amylase with yields of 96% directly from the fermentation broth.",
publisher = "Nature Publishing Group, London",
journal = "Scientific Reports",
title = "Mixed-mode resins: taking shortcut in downstream processing of raw-starch digesting alpha-amylases",
volume = "5",
doi = "10.1038/srep15772"
}

Lončar, N. L., Šokarda-Slavić, M., Vujčić, Z.,& Božić, N.. (2015). Mixed-mode resins: taking shortcut in downstream processing of raw-starch digesting alpha-amylases. in Scientific Reports
Nature Publishing Group, London., 5.
https://doi.org/10.1038/srep15772

Lončar NL, Šokarda-Slavić M, Vujčić Z, Božić N. Mixed-mode resins: taking shortcut in downstream processing of raw-starch digesting alpha-amylases. in Scientific Reports. 2015;5.
doi:10.1038/srep15772 .

Lončar, Nikola L., Šokarda-Slavić, Marinela, Vujčić, Zoran, Božić, Nataša, "Mixed-mode resins: taking shortcut in downstream processing of raw-starch digesting alpha-amylases" in Scientific Reports, 5 (2015),
https://doi.org/10.1038/srep15772 . .

TY  - DATA
AU  - Lončar, Nikola L.
AU  - Šokarda-Slavić, Marinela
AU  - Vujčić, Zoran
AU  - Božić, Nataša
PY  - 2015
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3384
PB  - Nature Publishing Group, London
T2  - Scientific Reports
T1  - Supplementary data for article: Lončar, N.; Slavić, M. Š.; Vujčić, Z.; Božić, N. Mixed-Mode Resins: Taking Shortcut in Downstream Processing of Raw-Starch Digesting α-Amylases. Scientific Reports 2015, 5. https://doi.org/10.1038/srep15772
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3384
ER  - 

@misc{
author = "Lončar, Nikola L. and Šokarda-Slavić, Marinela and Vujčić, Zoran and Božić, Nataša",
year = "2015",
publisher = "Nature Publishing Group, London",
journal = "Scientific Reports",
title = "Supplementary data for article: Lončar, N.; Slavić, M. Š.; Vujčić, Z.; Božić, N. Mixed-Mode Resins: Taking Shortcut in Downstream Processing of Raw-Starch Digesting α-Amylases. Scientific Reports 2015, 5. https://doi.org/10.1038/srep15772",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3384"
}

Lončar, N. L., Šokarda-Slavić, M., Vujčić, Z.,& Božić, N.. (2015). Supplementary data for article: Lončar, N.; Slavić, M. Š.; Vujčić, Z.; Božić, N. Mixed-Mode Resins: Taking Shortcut in Downstream Processing of Raw-Starch Digesting α-Amylases. Scientific Reports 2015, 5. https://doi.org/10.1038/srep15772. in Scientific Reports
Nature Publishing Group, London..
https://hdl.handle.net/21.15107/rcub_cherry_3384

Lončar NL, Šokarda-Slavić M, Vujčić Z, Božić N. Supplementary data for article: Lončar, N.; Slavić, M. Š.; Vujčić, Z.; Božić, N. Mixed-Mode Resins: Taking Shortcut in Downstream Processing of Raw-Starch Digesting α-Amylases. Scientific Reports 2015, 5. https://doi.org/10.1038/srep15772. in Scientific Reports. 2015;.
https://hdl.handle.net/21.15107/rcub_cherry_3384 .

Lončar, Nikola L., Šokarda-Slavić, Marinela, Vujčić, Zoran, Božić, Nataša, "Supplementary data for article: Lončar, N.; Slavić, M. Š.; Vujčić, Z.; Božić, N. Mixed-Mode Resins: Taking Shortcut in Downstream Processing of Raw-Starch Digesting α-Amylases. Scientific Reports 2015, 5. https://doi.org/10.1038/srep15772" in Scientific Reports (2015),
https://hdl.handle.net/21.15107/rcub_cherry_3384 .

TY  - JOUR
AU  - Gligorijević, Nikola
AU  - Stevanović, Nikola R.
AU  - Lončar, Nikola L.
AU  - Baošić, Rada
AU  - Vujčić, Zoran
AU  - Božić, Nataša
PY  - 2014
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1771
AB  - Several natural isolates of Bacillus strains namely 5B, 12B, 16B, 18 and 24B were grown at two different temperatures in submerged fermentation for the production of raw-starch-digesting alpha-amylases. All strains except Bacillus sp. 18 produced more alpha-amylase at 37 degrees C. The hydrolysis of raw cornstarch followed the same pattern. Efficient hydrolysis was obtained with alpha-amylases from Bacillus sp. 5B, 12B, 16B and 24B grown at 37 degrees C and Bacillus sp. 18 grown at 50 degrees C. Zymography after isoelectric focusing showed that alpha-amylases were produced in multiple forms, from 2 to 6, depending on the strain when they were growing at 37 degrees C, while growth at 50 degrees C induced only 1 or 2 isoforms. Thin layer chromatography (TLC) analysis of the hydrolysis products of raw corn and soluble starch by alpha-amylases revealed the production of various mixtures of oligosaccharides. In most cases, G3 was the most dominant product from soluble starch while G2, G3 and G5 were the main products of raw starch hydrolysis. This indicates that the obtained alpha-amylases could be used for starch liquidification or short-chain-oligosaccharide formation, depending on the type of starch (raw or soluble) used for the hydrolysis.
PB  - Serbian Chemical Soc, Belgrade
T2  - Journal of the Serbian Chemical Society
T1  - A thin layer chromatographic comparison of raw and soluble starch hydrolysis patterns of some alpha-amylases from Bacillus sp isolated in Serbia
VL  - 79
IS  - 4
SP  - 411
EP  - 420
DO  - 10.2298/JSC130909155G
ER  - 

@article{
author = "Gligorijević, Nikola and Stevanović, Nikola R. and Lončar, Nikola L. and Baošić, Rada and Vujčić, Zoran and Božić, Nataša",
year = "2014",
abstract = "Several natural isolates of Bacillus strains namely 5B, 12B, 16B, 18 and 24B were grown at two different temperatures in submerged fermentation for the production of raw-starch-digesting alpha-amylases. All strains except Bacillus sp. 18 produced more alpha-amylase at 37 degrees C. The hydrolysis of raw cornstarch followed the same pattern. Efficient hydrolysis was obtained with alpha-amylases from Bacillus sp. 5B, 12B, 16B and 24B grown at 37 degrees C and Bacillus sp. 18 grown at 50 degrees C. Zymography after isoelectric focusing showed that alpha-amylases were produced in multiple forms, from 2 to 6, depending on the strain when they were growing at 37 degrees C, while growth at 50 degrees C induced only 1 or 2 isoforms. Thin layer chromatography (TLC) analysis of the hydrolysis products of raw corn and soluble starch by alpha-amylases revealed the production of various mixtures of oligosaccharides. In most cases, G3 was the most dominant product from soluble starch while G2, G3 and G5 were the main products of raw starch hydrolysis. This indicates that the obtained alpha-amylases could be used for starch liquidification or short-chain-oligosaccharide formation, depending on the type of starch (raw or soluble) used for the hydrolysis.",
publisher = "Serbian Chemical Soc, Belgrade",
journal = "Journal of the Serbian Chemical Society",
title = "A thin layer chromatographic comparison of raw and soluble starch hydrolysis patterns of some alpha-amylases from Bacillus sp isolated in Serbia",
volume = "79",
number = "4",
pages = "411-420",
doi = "10.2298/JSC130909155G"
}

Gligorijević, N., Stevanović, N. R., Lončar, N. L., Baošić, R., Vujčić, Z.,& Božić, N.. (2014). A thin layer chromatographic comparison of raw and soluble starch hydrolysis patterns of some alpha-amylases from Bacillus sp isolated in Serbia. in Journal of the Serbian Chemical Society
Serbian Chemical Soc, Belgrade., 79(4), 411-420.
https://doi.org/10.2298/JSC130909155G

Gligorijević N, Stevanović NR, Lončar NL, Baošić R, Vujčić Z, Božić N. A thin layer chromatographic comparison of raw and soluble starch hydrolysis patterns of some alpha-amylases from Bacillus sp isolated in Serbia. in Journal of the Serbian Chemical Society. 2014;79(4):411-420.
doi:10.2298/JSC130909155G .

Gligorijević, Nikola, Stevanović, Nikola R., Lončar, Nikola L., Baošić, Rada, Vujčić, Zoran, Božić, Nataša, "A thin layer chromatographic comparison of raw and soluble starch hydrolysis patterns of some alpha-amylases from Bacillus sp isolated in Serbia" in Journal of the Serbian Chemical Society, 79, no. 4 (2014):411-420,
https://doi.org/10.2298/JSC130909155G . .

TY  - JOUR
AU  - Lončar, Nikola L.
AU  - Gligorijević, Nikola
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2014
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1791
AB  - Halotolerant strains of Bacillus amyloliquefaciens were isolated from salt spring in Ovca spa located in Republic of Serbia. Strains exhibit robust spore laccase with high temperature optimum of 65 degrees C while pH optimum is wide and substrate dependant. Ability to oxidize azo dyes was demonstrated. Under optimized conditions more than 85% removal of Congo red dye was achieved at pH 5.7. Substantial resistance to inhibition by high concentration of chloride ions was observed and tolerance of some commonly used cosolvents shows that applicability of these laccases goes beyond decolorization of textile effluents. (C) 2014 Elsevier Ltd. All rights reserved.
PB  - Elsevier Sci Ltd, Oxford
T2  - International Biodeterioration and Biodegradation
T1  - Congo red degrading laccases from Bacillus amyloliquefaciens strains isolated from salt spring in Serbia
VL  - 91
SP  - 18
EP  - 23
DO  - 10.1016/j.ibiod.2014.03.008
ER  - 

@article{
author = "Lončar, Nikola L. and Gligorijević, Nikola and Božić, Nataša and Vujčić, Zoran",
year = "2014",
abstract = "Halotolerant strains of Bacillus amyloliquefaciens were isolated from salt spring in Ovca spa located in Republic of Serbia. Strains exhibit robust spore laccase with high temperature optimum of 65 degrees C while pH optimum is wide and substrate dependant. Ability to oxidize azo dyes was demonstrated. Under optimized conditions more than 85% removal of Congo red dye was achieved at pH 5.7. Substantial resistance to inhibition by high concentration of chloride ions was observed and tolerance of some commonly used cosolvents shows that applicability of these laccases goes beyond decolorization of textile effluents. (C) 2014 Elsevier Ltd. All rights reserved.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "International Biodeterioration and Biodegradation",
title = "Congo red degrading laccases from Bacillus amyloliquefaciens strains isolated from salt spring in Serbia",
volume = "91",
pages = "18-23",
doi = "10.1016/j.ibiod.2014.03.008"
}

Lončar, N. L., Gligorijević, N., Božić, N.,& Vujčić, Z.. (2014). Congo red degrading laccases from Bacillus amyloliquefaciens strains isolated from salt spring in Serbia. in International Biodeterioration and Biodegradation
Elsevier Sci Ltd, Oxford., 91, 18-23.
https://doi.org/10.1016/j.ibiod.2014.03.008

Lončar NL, Gligorijević N, Božić N, Vujčić Z. Congo red degrading laccases from Bacillus amyloliquefaciens strains isolated from salt spring in Serbia. in International Biodeterioration and Biodegradation. 2014;91:18-23.
doi:10.1016/j.ibiod.2014.03.008 .

Lončar, Nikola L., Gligorijević, Nikola, Božić, Nataša, Vujčić, Zoran, "Congo red degrading laccases from Bacillus amyloliquefaciens strains isolated from salt spring in Serbia" in International Biodeterioration and Biodegradation, 91 (2014):18-23,
https://doi.org/10.1016/j.ibiod.2014.03.008 . .

TY  - JOUR
AU  - Božić, Nataša
AU  - Šokarda-Slavić, Marinela
AU  - Gavrilović, Anja
AU  - Vujčić, Zoran
PY  - 2014
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1798
AB  - alpha-Amylase production by solid-state fermentation of different Bacillus sp. was studied previously on different fermentation media. However, no study has been reported on the influence of selected media on expression of desired amylase isoforms such as raw-starch-digesting amylase (RSDA). In this paper, the influence of different inexpensive and available agro-resources as solid media (corn, wheat and triticale) on alpha-amylase isoform induction from three wild-type Bacillus sp., selected among one hundred strains tested, namely 9B, 12B and 24A was investigated. For all three strains, tested amylases were detected in the multiple forms; however, number and intensity of each form differed depending on the solid media used for growth. To determine which isoform from Bacillus sp. 12B was RSDA, the suspected isoform was purified. The optimum pH for the purified alpha-amylase isoform was 6.0-8.0, while the optimum temperature was 60-90 A degrees C. Isoform was considerably thermostable and Ca2+-independent, and actually the only alpha-amylase active towards raw starch. Purification and characterization of RSDA showed that not all of the solid media tested induced RSDA. From an economic point of view, it might be significant to obtain pure isoenzyme for potential use in the raw-starch hydrolysis, since it was 5 times more efficient in raw corn starch hydrolysis than the crude amylase preparation.
PB  - Springer, New York
T2  - Bioprocess and Biosystems Engineering
T1  - Production of raw-starch-digesting alpha-amylase isoform from Bacillus sp under solid-state fermentation and biochemical characterization
VL  - 37
IS  - 7
SP  - 1353
EP  - 1360
DO  - 10.1007/s00449-013-1105-1
ER  - 

@article{
author = "Božić, Nataša and Šokarda-Slavić, Marinela and Gavrilović, Anja and Vujčić, Zoran",
year = "2014",
abstract = "alpha-Amylase production by solid-state fermentation of different Bacillus sp. was studied previously on different fermentation media. However, no study has been reported on the influence of selected media on expression of desired amylase isoforms such as raw-starch-digesting amylase (RSDA). In this paper, the influence of different inexpensive and available agro-resources as solid media (corn, wheat and triticale) on alpha-amylase isoform induction from three wild-type Bacillus sp., selected among one hundred strains tested, namely 9B, 12B and 24A was investigated. For all three strains, tested amylases were detected in the multiple forms; however, number and intensity of each form differed depending on the solid media used for growth. To determine which isoform from Bacillus sp. 12B was RSDA, the suspected isoform was purified. The optimum pH for the purified alpha-amylase isoform was 6.0-8.0, while the optimum temperature was 60-90 A degrees C. Isoform was considerably thermostable and Ca2+-independent, and actually the only alpha-amylase active towards raw starch. Purification and characterization of RSDA showed that not all of the solid media tested induced RSDA. From an economic point of view, it might be significant to obtain pure isoenzyme for potential use in the raw-starch hydrolysis, since it was 5 times more efficient in raw corn starch hydrolysis than the crude amylase preparation.",
publisher = "Springer, New York",
journal = "Bioprocess and Biosystems Engineering",
title = "Production of raw-starch-digesting alpha-amylase isoform from Bacillus sp under solid-state fermentation and biochemical characterization",
volume = "37",
number = "7",
pages = "1353-1360",
doi = "10.1007/s00449-013-1105-1"
}

Božić, N., Šokarda-Slavić, M., Gavrilović, A.,& Vujčić, Z.. (2014). Production of raw-starch-digesting alpha-amylase isoform from Bacillus sp under solid-state fermentation and biochemical characterization. in Bioprocess and Biosystems Engineering
Springer, New York., 37(7), 1353-1360.
https://doi.org/10.1007/s00449-013-1105-1

Božić N, Šokarda-Slavić M, Gavrilović A, Vujčić Z. Production of raw-starch-digesting alpha-amylase isoform from Bacillus sp under solid-state fermentation and biochemical characterization. in Bioprocess and Biosystems Engineering. 2014;37(7):1353-1360.
doi:10.1007/s00449-013-1105-1 .

Božić, Nataša, Šokarda-Slavić, Marinela, Gavrilović, Anja, Vujčić, Zoran, "Production of raw-starch-digesting alpha-amylase isoform from Bacillus sp under solid-state fermentation and biochemical characterization" in Bioprocess and Biosystems Engineering, 37, no. 7 (2014):1353-1360,
https://doi.org/10.1007/s00449-013-1105-1 . .

TY  - JOUR
AU  - Pešić, Milja
AU  - Božić, Nataša
AU  - Lopez, Carmen
AU  - Lončar, Nikola L.
AU  - Alvaro, Gregorio
AU  - Vujčić, Zoran
PY  - 2014
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1844
AB  - Chloroperoxidase from Caldariomyces fumago (CPO, EC 1.11.1.10) is one of the most interesting enzymes from the group of heme peroxidases and has been extensively applied in synthetic processes. Nevertheless, the practical application of CPO is limited due to its very low operational stability, especially in the presence of peroxidative compounds. For this reason, effect of chemical modifications of CPO in the stability of the enzyme was studied. Side-chain selective modifications of amino groups of Lys residues, and carboxyl groups of Asp and Glu residues, as well as crosslinking and periodate oxidation of sugar moiety were carried out. The stability of modified CPOs was evaluated at elevated pH and temperature, and in the presence of tert-butyl hydroperoxide. Effect of modification of CPO on the performance of the reaction of Cbz-ethanolamine oxidation was studied as well. Those modifications that involved carboxyl groups via carbodiimide coupled method and the periodate oxidation of the sugar moiety produced better catalysts than native CPO in terms of stability and activity at elevated pH values and temperatures. (C) 2014 Elsevier Ltd. All rights reserved.
PB  - Elsevier Sci Ltd, Oxford
T2  - Process Biochemistry
T1  - Chemical modification of chloroperoxidase for enhanced stability and activity
VL  - 49
IS  - 9
SP  - 1472
EP  - 1479
DO  - 10.1016/j.procbio.2014.05.025
ER  - 

@article{
author = "Pešić, Milja and Božić, Nataša and Lopez, Carmen and Lončar, Nikola L. and Alvaro, Gregorio and Vujčić, Zoran",
year = "2014",
abstract = "Chloroperoxidase from Caldariomyces fumago (CPO, EC 1.11.1.10) is one of the most interesting enzymes from the group of heme peroxidases and has been extensively applied in synthetic processes. Nevertheless, the practical application of CPO is limited due to its very low operational stability, especially in the presence of peroxidative compounds. For this reason, effect of chemical modifications of CPO in the stability of the enzyme was studied. Side-chain selective modifications of amino groups of Lys residues, and carboxyl groups of Asp and Glu residues, as well as crosslinking and periodate oxidation of sugar moiety were carried out. The stability of modified CPOs was evaluated at elevated pH and temperature, and in the presence of tert-butyl hydroperoxide. Effect of modification of CPO on the performance of the reaction of Cbz-ethanolamine oxidation was studied as well. Those modifications that involved carboxyl groups via carbodiimide coupled method and the periodate oxidation of the sugar moiety produced better catalysts than native CPO in terms of stability and activity at elevated pH values and temperatures. (C) 2014 Elsevier Ltd. All rights reserved.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Process Biochemistry",
title = "Chemical modification of chloroperoxidase for enhanced stability and activity",
volume = "49",
number = "9",
pages = "1472-1479",
doi = "10.1016/j.procbio.2014.05.025"
}

Pešić, M., Božić, N., Lopez, C., Lončar, N. L., Alvaro, G.,& Vujčić, Z.. (2014). Chemical modification of chloroperoxidase for enhanced stability and activity. in Process Biochemistry
Elsevier Sci Ltd, Oxford., 49(9), 1472-1479.
https://doi.org/10.1016/j.procbio.2014.05.025

Pešić M, Božić N, Lopez C, Lončar NL, Alvaro G, Vujčić Z. Chemical modification of chloroperoxidase for enhanced stability and activity. in Process Biochemistry. 2014;49(9):1472-1479.
doi:10.1016/j.procbio.2014.05.025 .

Pešić, Milja, Božić, Nataša, Lopez, Carmen, Lončar, Nikola L., Alvaro, Gregorio, Vujčić, Zoran, "Chemical modification of chloroperoxidase for enhanced stability and activity" in Process Biochemistry, 49, no. 9 (2014):1472-1479,
https://doi.org/10.1016/j.procbio.2014.05.025 . .