TY  - JOUR
AU  - Kosić, Višnja
AU  - Božić, Nataša
AU  - Dojnov, Biljana
AU  - Banković, Predrag
AU  - Jović-Jovičić, Nataša
AU  - Knežević-Jugović, Zorica
AU  - Milutinović Nikolić, Aleksandra
PY  - 2024
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7525
AB  - Three glycoside hydrolases (α-amylase, xylanase and pullulanase) were immobilized on low-cost, environmentally friendly, easily modified clay rich in beidellite. Modifications included common procedures: Na-exchange, acid activation, pillaring, pillaring followed by acid activation, and organo-modifications with chitosan. Supports were characterized by chemical analysis, low temperature N2 physisorption, X-ray powder diffraction (XRPD) and Fourier-transform infrared (FT-IR) spectroscopy. The point of zero charge was also determined. Specific activity of different immobilizates of selected glycoside hydrolases was notably influenced by the type of chemical modification of supports. For each enzyme optimal support was chosen and storage stability was tested. α-Amylase immobilized on acid-activated support retained up to 95% of its initial specific activity of 105.6 ± 5.1 U g−1 after a testing period of 120 days. The most suitable support for xylanase was chitosan-modified beidellite with having specific activity of 90.0 ± 1.4 U g−1 which retained >50% its value after 120 days. Specific activity of pullulanase immobilized on pillared sample that was subsequently activated by acid was 44.5 ± 0.7 U g−1. Initial activity was preserved up to 33% for the same testing period. Comparing these results to the storage stability of the free enzymes that completely lost their activity for the longest period of 40 days, it can be concluded that appropriately modified beidellite- based clays could be used as suitable supports for stabilization of glycoside hydrolases. Nevertheless, further characterization of immobilizates (pH, thermal and operational stability) is needed in order to raise the suitability for larger scale processes in food industry.
PB  - Elsevier
T2  - Applied Clay Science
T1  - Significantly improved stabilization of glycoside hydrolases important in food industry by immobilization onto appropriately modified beidellite
VL  - 250
SP  - 107289
DO  - 10.1016/j.clay.2024.107289
ER  - 

@article{
author = "Kosić, Višnja and Božić, Nataša and Dojnov, Biljana and Banković, Predrag and Jović-Jovičić, Nataša and Knežević-Jugović, Zorica and Milutinović Nikolić, Aleksandra",
year = "2024",
abstract = "Three glycoside hydrolases (α-amylase, xylanase and pullulanase) were immobilized on low-cost, environmentally friendly, easily modified clay rich in beidellite. Modifications included common procedures: Na-exchange, acid activation, pillaring, pillaring followed by acid activation, and organo-modifications with chitosan. Supports were characterized by chemical analysis, low temperature N2 physisorption, X-ray powder diffraction (XRPD) and Fourier-transform infrared (FT-IR) spectroscopy. The point of zero charge was also determined. Specific activity of different immobilizates of selected glycoside hydrolases was notably influenced by the type of chemical modification of supports. For each enzyme optimal support was chosen and storage stability was tested. α-Amylase immobilized on acid-activated support retained up to 95% of its initial specific activity of 105.6 ± 5.1 U g−1 after a testing period of 120 days. The most suitable support for xylanase was chitosan-modified beidellite with having specific activity of 90.0 ± 1.4 U g−1 which retained >50% its value after 120 days. Specific activity of pullulanase immobilized on pillared sample that was subsequently activated by acid was 44.5 ± 0.7 U g−1. Initial activity was preserved up to 33% for the same testing period. Comparing these results to the storage stability of the free enzymes that completely lost their activity for the longest period of 40 days, it can be concluded that appropriately modified beidellite- based clays could be used as suitable supports for stabilization of glycoside hydrolases. Nevertheless, further characterization of immobilizates (pH, thermal and operational stability) is needed in order to raise the suitability for larger scale processes in food industry.",
publisher = "Elsevier",
journal = "Applied Clay Science",
title = "Significantly improved stabilization of glycoside hydrolases important in food industry by immobilization onto appropriately modified beidellite",
volume = "250",
pages = "107289",
doi = "10.1016/j.clay.2024.107289"
}

Kosić, V., Božić, N., Dojnov, B., Banković, P., Jović-Jovičić, N., Knežević-Jugović, Z.,& Milutinović Nikolić, A.. (2024). Significantly improved stabilization of glycoside hydrolases important in food industry by immobilization onto appropriately modified beidellite. in Applied Clay Science
Elsevier., 250, 107289.
https://doi.org/10.1016/j.clay.2024.107289

Kosić V, Božić N, Dojnov B, Banković P, Jović-Jovičić N, Knežević-Jugović Z, Milutinović Nikolić A. Significantly improved stabilization of glycoside hydrolases important in food industry by immobilization onto appropriately modified beidellite. in Applied Clay Science. 2024;250:107289.
doi:10.1016/j.clay.2024.107289 .

Kosić, Višnja, Božić, Nataša, Dojnov, Biljana, Banković, Predrag, Jović-Jovičić, Nataša, Knežević-Jugović, Zorica, Milutinović Nikolić, Aleksandra, "Significantly improved stabilization of glycoside hydrolases important in food industry by immobilization onto appropriately modified beidellite" in Applied Clay Science, 250 (2024):107289,
https://doi.org/10.1016/j.clay.2024.107289 . .

TY  - CONF
AU  - Kosić, Višnja
AU  - Božić, Nataša
AU  - Dojnov, Biljana
AU  - Stevanović, Gordana
AU  - Banković, Predrag
AU  - Milutinović Nikolić, Aleksandra
AU  - Knežević-Jugović, Zorica
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6846
AB  - Clays are naturally occurring, environmentally friendly, chemically inert, thermostable, inexpensive resources that are easily modified into materials with tailored properties. As such, they can be used as suitable supports for enzyme immobilization and application in the food industry. Natural polysaccharides starch, xylan, pullulan, and its derivatives obtained by the action of enzymes, have numerous potentials for food industrial applications. In this work the enzyme supports were prepared from bentonite from Coal mine "Bogovina", Serbia by acid activation (AA), pillaring (P), and pillaring followed by acid activation (PAA). The characterization of the obtained materials included chemical and phase composition, surface acidity, and textural properties. After characterization, -amylase from Bacillus paralicheniformis (BliAmy), commercial xylanase from Sigma-Aldrich (Xyl), and pullulanase from B. paralicheniformis (BliPull) were immobilized on bentonite based supports by 24 h adsorption at 25 °C. The obtained biocatalysts BliAmy-AA (106 IU/g), Xyl-P (74 IU/g), and BliPull-PAA (45 IU/g) showed very good storage stability with the activity preserved after 4 weeks of testing. Products of hydrolysis were detected by TLC and indicate a promising application in the food industry.
PB  - Belgrade : Serbian Chemical Society
C3  - Abstract Book - XXII Congress EuroFoodChem, June 14-16, 2023, Belgrade, Serbia, 2023, 210-210
T1  - Stable, environmentally friendly and inexpensive biocatalysts for obtaining important ingredients applicable in the food industry
SP  - 210
EP  - 210
UR  - https://hdl.handle.net/21.15107/rcub_cer_6846
ER  - 

@conference{
author = "Kosić, Višnja and Božić, Nataša and Dojnov, Biljana and Stevanović, Gordana and Banković, Predrag and Milutinović Nikolić, Aleksandra and Knežević-Jugović, Zorica",
year = "2023",
abstract = "Clays are naturally occurring, environmentally friendly, chemically inert, thermostable, inexpensive resources that are easily modified into materials with tailored properties. As such, they can be used as suitable supports for enzyme immobilization and application in the food industry. Natural polysaccharides starch, xylan, pullulan, and its derivatives obtained by the action of enzymes, have numerous potentials for food industrial applications. In this work the enzyme supports were prepared from bentonite from Coal mine "Bogovina", Serbia by acid activation (AA), pillaring (P), and pillaring followed by acid activation (PAA). The characterization of the obtained materials included chemical and phase composition, surface acidity, and textural properties. After characterization, -amylase from Bacillus paralicheniformis (BliAmy), commercial xylanase from Sigma-Aldrich (Xyl), and pullulanase from B. paralicheniformis (BliPull) were immobilized on bentonite based supports by 24 h adsorption at 25 °C. The obtained biocatalysts BliAmy-AA (106 IU/g), Xyl-P (74 IU/g), and BliPull-PAA (45 IU/g) showed very good storage stability with the activity preserved after 4 weeks of testing. Products of hydrolysis were detected by TLC and indicate a promising application in the food industry.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "Abstract Book - XXII Congress EuroFoodChem, June 14-16, 2023, Belgrade, Serbia, 2023, 210-210",
title = "Stable, environmentally friendly and inexpensive biocatalysts for obtaining important ingredients applicable in the food industry",
pages = "210-210",
url = "https://hdl.handle.net/21.15107/rcub_cer_6846"
}

Kosić, V., Božić, N., Dojnov, B., Stevanović, G., Banković, P., Milutinović Nikolić, A.,& Knežević-Jugović, Z.. (2023). Stable, environmentally friendly and inexpensive biocatalysts for obtaining important ingredients applicable in the food industry. in Abstract Book - XXII Congress EuroFoodChem, June 14-16, 2023, Belgrade, Serbia, 2023, 210-210
Belgrade : Serbian Chemical Society., 210-210.
https://hdl.handle.net/21.15107/rcub_cer_6846

Kosić V, Božić N, Dojnov B, Stevanović G, Banković P, Milutinović Nikolić A, Knežević-Jugović Z. Stable, environmentally friendly and inexpensive biocatalysts for obtaining important ingredients applicable in the food industry. in Abstract Book - XXII Congress EuroFoodChem, June 14-16, 2023, Belgrade, Serbia, 2023, 210-210. 2023;:210-210.
https://hdl.handle.net/21.15107/rcub_cer_6846 .

Kosić, Višnja, Božić, Nataša, Dojnov, Biljana, Stevanović, Gordana, Banković, Predrag, Milutinović Nikolić, Aleksandra, Knežević-Jugović, Zorica, "Stable, environmentally friendly and inexpensive biocatalysts for obtaining important ingredients applicable in the food industry" in Abstract Book - XXII Congress EuroFoodChem, June 14-16, 2023, Belgrade, Serbia, 2023, 210-210 (2023):210-210,
https://hdl.handle.net/21.15107/rcub_cer_6846 .

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  - https://cer.ihtm.bg.ac.rs/handle/123456789/5787
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.
PB  - Elsevier Ltd
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.",
publisher = "Elsevier Ltd",
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
Elsevier Ltd., 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  - CONF
AU  - Ristović, Marina
AU  - Stojanović, Sanja
AU  - Šokarda Slavić, Marinela
AU  - Margetić, Aleksandra
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Dojnov, Biljana
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6029
AB  - Since each region of the world has different agricultural sectors, specific residues are generating and at the same time a wide range of alternative novel sources of nutrients emerge, such as carbohydrates, proteins and minerals. Usage of biomass residues as start material for prebiotics production is preferable from the standpoint of ecology and as cheap production process. Corn cob can be considered as the main source for xylooligosaccharides (XOS) production. XOS are prebiotic, functional food ingredients, with biological benefits such as immunomodulatory and anti-inflammatory properties, anticancer and antioxidant activity. The European Commission has approved XOS produced from corn cob as a safe for human consumption. The corn cob mainly consists of lignocellulose. Chemical treatment is used to extract the hemicelullose from corn cob which is then hydrolyzed to XOS by fungal xylanases. Xylanase produced by fungi genera Aspergillus and Trichoderma are considered key enzymes for XOS production. Fermentation enzyme extract of strain Aspergillus tubingensis FAT35, obtained after SSF on corn cob as substrate, was used for hydrolysis of corn cob xylan to XOS. A. tubigensis FAT35 produced high level of xylanase (350 U/mL). Obtained XOS (2-10 units) were characterized by TLC and by antioxidant activity (ORAC, DPPH, FRAP, and ABTS). Significant antioxidant potential was shown by all used antioxidant tests. The obtained XOS are suitable to be a functional food additive and are obtained from agro-waste material by environmentally and economically suitable way.
PB  - European Federation of Biotechnology
C3  - Programme and abstract book - Biotechnology for a circular bioeconomy: carbon capture, waste recycling and mitigation of global warming, 28-29 March 2023, online
T1  - Corn cob agro-waste as valuable material for XOS production by fungal enzymes
SP  - 67
EP  - 67
UR  - https://hdl.handle.net/21.15107/rcub_cer_6029
ER  - 

@conference{
author = "Ristović, Marina and Stojanović, Sanja and Šokarda Slavić, Marinela and Margetić, Aleksandra and Božić, Nataša and Vujčić, Zoran and Dojnov, Biljana",
year = "2023",
abstract = "Since each region of the world has different agricultural sectors, specific residues are generating and at the same time a wide range of alternative novel sources of nutrients emerge, such as carbohydrates, proteins and minerals. Usage of biomass residues as start material for prebiotics production is preferable from the standpoint of ecology and as cheap production process. Corn cob can be considered as the main source for xylooligosaccharides (XOS) production. XOS are prebiotic, functional food ingredients, with biological benefits such as immunomodulatory and anti-inflammatory properties, anticancer and antioxidant activity. The European Commission has approved XOS produced from corn cob as a safe for human consumption. The corn cob mainly consists of lignocellulose. Chemical treatment is used to extract the hemicelullose from corn cob which is then hydrolyzed to XOS by fungal xylanases. Xylanase produced by fungi genera Aspergillus and Trichoderma are considered key enzymes for XOS production. Fermentation enzyme extract of strain Aspergillus tubingensis FAT35, obtained after SSF on corn cob as substrate, was used for hydrolysis of corn cob xylan to XOS. A. tubigensis FAT35 produced high level of xylanase (350 U/mL). Obtained XOS (2-10 units) were characterized by TLC and by antioxidant activity (ORAC, DPPH, FRAP, and ABTS). Significant antioxidant potential was shown by all used antioxidant tests. The obtained XOS are suitable to be a functional food additive and are obtained from agro-waste material by environmentally and economically suitable way.",
publisher = "European Federation of Biotechnology",
journal = "Programme and abstract book - Biotechnology for a circular bioeconomy: carbon capture, waste recycling and mitigation of global warming, 28-29 March 2023, online",
title = "Corn cob agro-waste as valuable material for XOS production by fungal enzymes",
pages = "67-67",
url = "https://hdl.handle.net/21.15107/rcub_cer_6029"
}

Ristović, M., Stojanović, S., Šokarda Slavić, M., Margetić, A., Božić, N., Vujčić, Z.,& Dojnov, B.. (2023). Corn cob agro-waste as valuable material for XOS production by fungal enzymes. in Programme and abstract book - Biotechnology for a circular bioeconomy: carbon capture, waste recycling and mitigation of global warming, 28-29 March 2023, online
European Federation of Biotechnology., 67-67.
https://hdl.handle.net/21.15107/rcub_cer_6029

Ristović M, Stojanović S, Šokarda Slavić M, Margetić A, Božić N, Vujčić Z, Dojnov B. Corn cob agro-waste as valuable material for XOS production by fungal enzymes. in Programme and abstract book - Biotechnology for a circular bioeconomy: carbon capture, waste recycling and mitigation of global warming, 28-29 March 2023, online. 2023;:67-67.
https://hdl.handle.net/21.15107/rcub_cer_6029 .

Ristović, Marina, Stojanović, Sanja, Šokarda Slavić, Marinela, Margetić, Aleksandra, Božić, Nataša, Vujčić, Zoran, Dojnov, Biljana, "Corn cob agro-waste as valuable material for XOS production by fungal enzymes" in Programme and abstract book - Biotechnology for a circular bioeconomy: carbon capture, waste recycling and mitigation of global warming, 28-29 March 2023, online (2023):67-67,
https://hdl.handle.net/21.15107/rcub_cer_6029 .

TY  - CONF
AU  - Kosić, Višnja
AU  - Božić, Nataša
AU  - Dojnov, Biljana
AU  - Stevanović, Gordana
AU  - Milutinović Nikolić, Aleksandra
AU  - Knežević-Jugović, Zorica
AU  - Banković, Predrag
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6030
AB  - Some coal mines suffer from problem of having huge deposits of bentonite that they regard as undesirable, since bentonite can cause problem due to its swelling property. Instead of piling stacks of bentonite and regarding it as ecological problem the bentonite can be used as enzyme support by immobilization of enzymes as the challenging new application in the field of biotechnology. The enzyme catalysts in the industrial application present lower energy costs and are more environmentally friendly in comparison to traditional chemical processes. The advantages of enzymes are even more prominent when enzymes are applied in immobilized form. In this work the enzyme supports were prepared from bentonite from Coal mine "Bogovina", Serbia where bentonite, although valuable resource, is still considered to be tailings. Bentonite was modified by common methods: acid activation (AA), pillaring (P), and pillaring followed by acid activation (PAA) and tested as enzyme support. All the obtained materials were characterized by the X-ray powder diffraction and FTIR spectroscopy. The amylase, xylanase, and pullulanase were immobilized on different bentonite based supports by 24 h adsorption at 25 °C. The experimental results revealed that under the investigated conditions AA, P, and PAA, were the most suitable for amylase (106 IU/g), xylanase (74 IU/g), and pullulanase (45 IU/g) immobilization, respectively.
PB  - European Federation of Biotechnology
C3  - Programme and abstract book - Biotechnology for a circular bioeconomy: carbon capture, waste recycling and mitigation of global warming, 28-29 March 2023, online
T1  - The tailings from coal mine instead of waste became applicable as enzyme supports
SP  - 65
EP  - 65
UR  - https://hdl.handle.net/21.15107/rcub_cer_6030
ER  - 

@conference{
author = "Kosić, Višnja and Božić, Nataša and Dojnov, Biljana and Stevanović, Gordana and Milutinović Nikolić, Aleksandra and Knežević-Jugović, Zorica and Banković, Predrag",
year = "2023",
abstract = "Some coal mines suffer from problem of having huge deposits of bentonite that they regard as undesirable, since bentonite can cause problem due to its swelling property. Instead of piling stacks of bentonite and regarding it as ecological problem the bentonite can be used as enzyme support by immobilization of enzymes as the challenging new application in the field of biotechnology. The enzyme catalysts in the industrial application present lower energy costs and are more environmentally friendly in comparison to traditional chemical processes. The advantages of enzymes are even more prominent when enzymes are applied in immobilized form. In this work the enzyme supports were prepared from bentonite from Coal mine "Bogovina", Serbia where bentonite, although valuable resource, is still considered to be tailings. Bentonite was modified by common methods: acid activation (AA), pillaring (P), and pillaring followed by acid activation (PAA) and tested as enzyme support. All the obtained materials were characterized by the X-ray powder diffraction and FTIR spectroscopy. The amylase, xylanase, and pullulanase were immobilized on different bentonite based supports by 24 h adsorption at 25 °C. The experimental results revealed that under the investigated conditions AA, P, and PAA, were the most suitable for amylase (106 IU/g), xylanase (74 IU/g), and pullulanase (45 IU/g) immobilization, respectively.",
publisher = "European Federation of Biotechnology",
journal = "Programme and abstract book - Biotechnology for a circular bioeconomy: carbon capture, waste recycling and mitigation of global warming, 28-29 March 2023, online",
title = "The tailings from coal mine instead of waste became applicable as enzyme supports",
pages = "65-65",
url = "https://hdl.handle.net/21.15107/rcub_cer_6030"
}

Kosić, V., Božić, N., Dojnov, B., Stevanović, G., Milutinović Nikolić, A., Knežević-Jugović, Z.,& Banković, P.. (2023). The tailings from coal mine instead of waste became applicable as enzyme supports. in Programme and abstract book - Biotechnology for a circular bioeconomy: carbon capture, waste recycling and mitigation of global warming, 28-29 March 2023, online
European Federation of Biotechnology., 65-65.
https://hdl.handle.net/21.15107/rcub_cer_6030

Kosić V, Božić N, Dojnov B, Stevanović G, Milutinović Nikolić A, Knežević-Jugović Z, Banković P. The tailings from coal mine instead of waste became applicable as enzyme supports. in Programme and abstract book - Biotechnology for a circular bioeconomy: carbon capture, waste recycling and mitigation of global warming, 28-29 March 2023, online. 2023;:65-65.
https://hdl.handle.net/21.15107/rcub_cer_6030 .

Kosić, Višnja, Božić, Nataša, Dojnov, Biljana, Stevanović, Gordana, Milutinović Nikolić, Aleksandra, Knežević-Jugović, Zorica, Banković, Predrag, "The tailings from coal mine instead of waste became applicable as enzyme supports" in Programme and abstract book - Biotechnology for a circular bioeconomy: carbon capture, waste recycling and mitigation of global warming, 28-29 March 2023, online (2023):65-65,
https://hdl.handle.net/21.15107/rcub_cer_6030 .

TY  - CONF
AU  - Stojanović, Sanja
AU  - Margetić, Aleksandra
AU  - Šokarda Slavić, Marinela
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Dojnov, Biljana
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6242
AB  - Most of the functional oligosaccharides (OS) consist of monomers, present in varying degrees of polymerization (DP) ranging between 3 and 10 units. DP of inulin-type fructooligosaccharides has a great impact on fermentability and their
utilization by probiotic bacteria such is Bifidobacteria, thus they have a great impact on their health-promoting effect. Technological properties of fructooligosaccharides (FOS) can improve the physicochemical and sensory characteristics of food products, leading to their increased application in the food industry. It has been found that microbial endoinulinase plays an important role in production of inulin-type fructooligosaccharides. Aspergillus welwitschiae FAW1 strain has proven to be non-toxigenic with the absence of biosynthetic gene clusters for mycotoxins (ochratoxins and fumonisins) and therefore safe for use in food production [5]. Growing on the natural substrate, triticale (Triticosecale sp) FAW1 strain produced inulinase complex from which endoinulinase (InuA) was purified by chromatographic techniques. FOS was prepared by time-controlled hydrolysis of inulin. Monitoring kinetics and determining the amount of obtained FOS by TLC and HPLC methods led to a conclusion that FOS production by hydrolysis of inulin is kinetic dependent reaction. Depending on the reaction time, FOS with different compositions are obtained.The largest amount of produced FOS (DP 2-6) has been in 15-20 minutes of the reaction, where the resulting mixture contains small amount of mono- and disaccharides. The obtained FOS were characterized on antioxidant capacity. Produced FOS showed significant antioxidant potential according to ORAC method which classifies them as potent candidate as additives in functional food. Endoinulinase (InuA) form A. welvitscihae FAW1 considered as key enzyme in FOS preparation. The composition and lenght of the produced FOS can be varied by controlling the reaction time, depending on the needs of of the market and their eventual application.
PB  - Serbian Chemical Society
C3  - Abstract Book - XXII Congress EuroFoodChem, June 14-16, 2023, Belgrade, Serbia
T1  - Obtaining of FOS by controlled hydrolysis of inulin with Aspergillus welwitschiae FAW1 endoinulinase
SP  - 203
EP  - 203
UR  - https://hdl.handle.net/21.15107/rcub_cer_6242
ER  - 

@conference{
author = "Stojanović, Sanja and Margetić, Aleksandra and Šokarda Slavić, Marinela and Božić, Nataša and Vujčić, Zoran and Dojnov, Biljana",
year = "2023",
abstract = "Most of the functional oligosaccharides (OS) consist of monomers, present in varying degrees of polymerization (DP) ranging between 3 and 10 units. DP of inulin-type fructooligosaccharides has a great impact on fermentability and their
utilization by probiotic bacteria such is Bifidobacteria, thus they have a great impact on their health-promoting effect. Technological properties of fructooligosaccharides (FOS) can improve the physicochemical and sensory characteristics of food products, leading to their increased application in the food industry. It has been found that microbial endoinulinase plays an important role in production of inulin-type fructooligosaccharides. Aspergillus welwitschiae FAW1 strain has proven to be non-toxigenic with the absence of biosynthetic gene clusters for mycotoxins (ochratoxins and fumonisins) and therefore safe for use in food production [5]. Growing on the natural substrate, triticale (Triticosecale sp) FAW1 strain produced inulinase complex from which endoinulinase (InuA) was purified by chromatographic techniques. FOS was prepared by time-controlled hydrolysis of inulin. Monitoring kinetics and determining the amount of obtained FOS by TLC and HPLC methods led to a conclusion that FOS production by hydrolysis of inulin is kinetic dependent reaction. Depending on the reaction time, FOS with different compositions are obtained.The largest amount of produced FOS (DP 2-6) has been in 15-20 minutes of the reaction, where the resulting mixture contains small amount of mono- and disaccharides. The obtained FOS were characterized on antioxidant capacity. Produced FOS showed significant antioxidant potential according to ORAC method which classifies them as potent candidate as additives in functional food. Endoinulinase (InuA) form A. welvitscihae FAW1 considered as key enzyme in FOS preparation. The composition and lenght of the produced FOS can be varied by controlling the reaction time, depending on the needs of of the market and their eventual application.",
publisher = "Serbian Chemical Society",
journal = "Abstract Book - XXII Congress EuroFoodChem, June 14-16, 2023, Belgrade, Serbia",
title = "Obtaining of FOS by controlled hydrolysis of inulin with Aspergillus welwitschiae FAW1 endoinulinase",
pages = "203-203",
url = "https://hdl.handle.net/21.15107/rcub_cer_6242"
}

Stojanović, S., Margetić, A., Šokarda Slavić, M., Božić, N., Vujčić, Z.,& Dojnov, B.. (2023). Obtaining of FOS by controlled hydrolysis of inulin with Aspergillus welwitschiae FAW1 endoinulinase. in Abstract Book - XXII Congress EuroFoodChem, June 14-16, 2023, Belgrade, Serbia
Serbian Chemical Society., 203-203.
https://hdl.handle.net/21.15107/rcub_cer_6242

Stojanović S, Margetić A, Šokarda Slavić M, Božić N, Vujčić Z, Dojnov B. Obtaining of FOS by controlled hydrolysis of inulin with Aspergillus welwitschiae FAW1 endoinulinase. in Abstract Book - XXII Congress EuroFoodChem, June 14-16, 2023, Belgrade, Serbia. 2023;:203-203.
https://hdl.handle.net/21.15107/rcub_cer_6242 .

Stojanović, Sanja, Margetić, Aleksandra, Šokarda Slavić, Marinela, Božić, Nataša, Vujčić, Zoran, Dojnov, Biljana, "Obtaining of FOS by controlled hydrolysis of inulin with Aspergillus welwitschiae FAW1 endoinulinase" in Abstract Book - XXII Congress EuroFoodChem, June 14-16, 2023, Belgrade, Serbia (2023):203-203,
https://hdl.handle.net/21.15107/rcub_cer_6242 .

TY  - JOUR
AU  - Šokarda Slavić, Marinela
AU  - Kojić, Milan
AU  - Margetić, Aleksandra
AU  - Stanisavljević, Nemanja
AU  - Gardijan, Lazar
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6458
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 = "Šokarda Slavić, Marinela and Kojić, Milan and Margetić, Aleksandra and Stanisavljević, Nemanja 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"
}

Šokarda Slavić, M., Kojić, M., Margetić, A., Stanisavljević, N., 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

Šokarda Slavić M, Kojić M, Margetić A, Stanisavljević N, 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 .

Šokarda Slavić, Marinela, Kojić, Milan, Margetić, Aleksandra, Stanisavljević, Nemanja, 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  - CONF
AU  - Ristović, Marina
AU  - Margetić, Aleksandra
AU  - Stojanović, Sanja
AU  - Šokarda Slavić, Marinela
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Dojnov, Biljana
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7135
AB  - Corn is widely used in the diet all over the world which generate corn cob as one of the most common agrowaste material. Corn cob mainly consists of lignocellulose with a high quantity of xylan; therefore, it can be considered the main source for xylooligosaccharides (XOS) production. XOS are prebiotic, functional food ingredients, which stand out on the market of oligosaccharide prebiotics because of their properties (antitumor effect, immunomodulatory effect, antioxidant power, and high pH and thermal stability). The European Commission has approved XOS produced from corn cob as safe for human consumption. For XOS production highly efficient xylanase obtained by Aspergillus tubingensis FAT35 under SSF on corn cob medium was used. Three methods of XOS production were evaluated: by hydrolysis of xylan alkali isolated from corn cob, by direct hydrolysis of xylan in corn cob and simultaneously production during fungal growth on corn cob (in situ). XOS were separated by ethanol precipitation from corn cob hydrolysate and from fermentation extract. In this way, the enzyme is concentrated in the precipitate, and XOS (remaining in the supernatant) are partially purified (separated from unhydrolyzed xylan and long chain oligomers). All prepared XOS were characterized by TLC and HPLC and antioxidant potential. Thanks to the high antioxidant potential (proven by DPPH, ABTS, FRAP, and ORAC tests) obtained XOS are good candidates for a functional food additive. Although the highest yield of XOS was obtained by the standard method (enzymatic hydrolysis of isolated xylan), additional methods represent the direct use of agro-waste material in an environmentally and economically suitable way. Special attention attracts in situ production because after isolation of XOS by ethanol precipitation the enzyme remains functional active and available for further use in next process.
PB  - Elsevier
C3  - Book of Abstracts - 3rd Food Chemistry Conference, 10-12 October 2023, Dresden, Germany
T1  - The use of corn cob as an excellent starting material for obtaining XOS and fungal enzymes involved in their production
SP  - P2.002
UR  - https://hdl.handle.net/21.15107/rcub_cer_7135
ER  - 

@conference{
author = "Ristović, Marina and Margetić, Aleksandra and Stojanović, Sanja and Šokarda Slavić, Marinela and Božić, Nataša and Vujčić, Zoran and Dojnov, Biljana",
year = "2023",
abstract = "Corn is widely used in the diet all over the world which generate corn cob as one of the most common agrowaste material. Corn cob mainly consists of lignocellulose with a high quantity of xylan; therefore, it can be considered the main source for xylooligosaccharides (XOS) production. XOS are prebiotic, functional food ingredients, which stand out on the market of oligosaccharide prebiotics because of their properties (antitumor effect, immunomodulatory effect, antioxidant power, and high pH and thermal stability). The European Commission has approved XOS produced from corn cob as safe for human consumption. For XOS production highly efficient xylanase obtained by Aspergillus tubingensis FAT35 under SSF on corn cob medium was used. Three methods of XOS production were evaluated: by hydrolysis of xylan alkali isolated from corn cob, by direct hydrolysis of xylan in corn cob and simultaneously production during fungal growth on corn cob (in situ). XOS were separated by ethanol precipitation from corn cob hydrolysate and from fermentation extract. In this way, the enzyme is concentrated in the precipitate, and XOS (remaining in the supernatant) are partially purified (separated from unhydrolyzed xylan and long chain oligomers). All prepared XOS were characterized by TLC and HPLC and antioxidant potential. Thanks to the high antioxidant potential (proven by DPPH, ABTS, FRAP, and ORAC tests) obtained XOS are good candidates for a functional food additive. Although the highest yield of XOS was obtained by the standard method (enzymatic hydrolysis of isolated xylan), additional methods represent the direct use of agro-waste material in an environmentally and economically suitable way. Special attention attracts in situ production because after isolation of XOS by ethanol precipitation the enzyme remains functional active and available for further use in next process.",
publisher = "Elsevier",
journal = "Book of Abstracts - 3rd Food Chemistry Conference, 10-12 October 2023, Dresden, Germany",
title = "The use of corn cob as an excellent starting material for obtaining XOS and fungal enzymes involved in their production",
pages = "P2.002",
url = "https://hdl.handle.net/21.15107/rcub_cer_7135"
}

Ristović, M., Margetić, A., Stojanović, S., Šokarda Slavić, M., Božić, N., Vujčić, Z.,& Dojnov, B.. (2023). The use of corn cob as an excellent starting material for obtaining XOS and fungal enzymes involved in their production. in Book of Abstracts - 3rd Food Chemistry Conference, 10-12 October 2023, Dresden, Germany
Elsevier., P2.002.
https://hdl.handle.net/21.15107/rcub_cer_7135

Ristović M, Margetić A, Stojanović S, Šokarda Slavić M, Božić N, Vujčić Z, Dojnov B. The use of corn cob as an excellent starting material for obtaining XOS and fungal enzymes involved in their production. in Book of Abstracts - 3rd Food Chemistry Conference, 10-12 October 2023, Dresden, Germany. 2023;:P2.002.
https://hdl.handle.net/21.15107/rcub_cer_7135 .

Ristović, Marina, Margetić, Aleksandra, Stojanović, Sanja, Šokarda Slavić, Marinela, Božić, Nataša, Vujčić, Zoran, Dojnov, Biljana, "The use of corn cob as an excellent starting material for obtaining XOS and fungal enzymes involved in their production" in Book of Abstracts - 3rd Food Chemistry Conference, 10-12 October 2023, Dresden, Germany (2023):P2.002,
https://hdl.handle.net/21.15107/rcub_cer_7135 .

TY  - CONF
AU  - Šokarda Slavić, Marinela
AU  - Margetić, Aleksandra
AU  - Dojnov, Biljana
AU  - Vujčić, Miroslava
AU  - Ristović, Marina
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7137
AB  - Given the growing concern about the depletion of fossil fuels, global warming, and the loss of natural resources, bioethanol made from sugar cane, molasses, and corn continues to garner interest globally and is regarded as the safest and cleanest alternative to oil. Starch is a widely available renewable carbohydrate from which bioethanol is conventionally obtained through energy demanding liquefaction and saccharification processes. A significant simplification of the process and a reduction of starch processing costs would be possible by applying raw starch hydrolysis using enzymes capable of degrading starch below the gelatinization temperature. A novel strategy for highly concentrated raw corn starch (30 % w/v) hydrolysis based on a modified simultaneous saccharification and fermentation process is optimized for the production of bioethanol. Different ratios of Bacillus paralicheniformis ATCC 9945a (BliAmy) and glucoamylase (Dextrozyme® GA), glucoamylase addition time, incubation time, and pH were investigated using a Box–Behnken experimental design to ensure high process efficiency. A two-step synergistic hydrolysis and fermentation with Saccharomyces cerevisiae at 30 °C was carried out in a single bioreactor vessel at the same pH (4.5). The obtained bioethanol concentration at 129.2 g/L, with a 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 indicates the viability of the proposed innovative process.
PB  - Slovenian Biochemical Society
C3  - Book of Abstracts - 15th Meeting of the Slovenian Biochemical Society with International Participation, 20-23 September 2023, Portorož, Slovenia
T1  - An innovative process for the production of bioethanol: Optimization and kinetic assessment
SP  - P59
UR  - https://hdl.handle.net/21.15107/rcub_cer_7137
ER  - 

@conference{
author = "Šokarda Slavić, Marinela and Margetić, Aleksandra and Dojnov, Biljana and Vujčić, Miroslava and Ristović, Marina and Božić, Nataša and Vujčić, Zoran",
year = "2023",
abstract = "Given the growing concern about the depletion of fossil fuels, global warming, and the loss of natural resources, bioethanol made from sugar cane, molasses, and corn continues to garner interest globally and is regarded as the safest and cleanest alternative to oil. Starch is a widely available renewable carbohydrate from which bioethanol is conventionally obtained through energy demanding liquefaction and saccharification processes. A significant simplification of the process and a reduction of starch processing costs would be possible by applying raw starch hydrolysis using enzymes capable of degrading starch below the gelatinization temperature. A novel strategy for highly concentrated raw corn starch (30 % w/v) hydrolysis based on a modified simultaneous saccharification and fermentation process is optimized for the production of bioethanol. Different ratios of Bacillus paralicheniformis ATCC 9945a (BliAmy) and glucoamylase (Dextrozyme® GA), glucoamylase addition time, incubation time, and pH were investigated using a Box–Behnken experimental design to ensure high process efficiency. A two-step synergistic hydrolysis and fermentation with Saccharomyces cerevisiae at 30 °C was carried out in a single bioreactor vessel at the same pH (4.5). The obtained bioethanol concentration at 129.2 g/L, with a 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 indicates the viability of the proposed innovative process.",
publisher = "Slovenian Biochemical Society",
journal = "Book of Abstracts - 15th Meeting of the Slovenian Biochemical Society with International Participation, 20-23 September 2023, Portorož, Slovenia",
title = "An innovative process for the production of bioethanol: Optimization and kinetic assessment",
pages = "P59",
url = "https://hdl.handle.net/21.15107/rcub_cer_7137"
}

Šokarda Slavić, M., Margetić, A., Dojnov, B., Vujčić, M., Ristović, M., Božić, N.,& Vujčić, Z.. (2023). An innovative process for the production of bioethanol: Optimization and kinetic assessment. in Book of Abstracts - 15th Meeting of the Slovenian Biochemical Society with International Participation, 20-23 September 2023, Portorož, Slovenia
Slovenian Biochemical Society., P59.
https://hdl.handle.net/21.15107/rcub_cer_7137

Šokarda Slavić M, Margetić A, Dojnov B, Vujčić M, Ristović M, Božić N, Vujčić Z. An innovative process for the production of bioethanol: Optimization and kinetic assessment. in Book of Abstracts - 15th Meeting of the Slovenian Biochemical Society with International Participation, 20-23 September 2023, Portorož, Slovenia. 2023;:P59.
https://hdl.handle.net/21.15107/rcub_cer_7137 .

Šokarda Slavić, Marinela, Margetić, Aleksandra, Dojnov, Biljana, Vujčić, Miroslava, Ristović, Marina, Božić, Nataša, Vujčić, Zoran, "An innovative process for the production of bioethanol: Optimization and kinetic assessment" in Book of Abstracts - 15th Meeting of the Slovenian Biochemical Society with International Participation, 20-23 September 2023, Portorož, Slovenia (2023):P59,
https://hdl.handle.net/21.15107/rcub_cer_7137 .

TY  - CONF
AU  - Stefanović, Marija
AU  - Savić, Aleksa
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Radosavljević, Jelena
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7345
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
SP  - 23
EP  - 23
UR  - https://hdl.handle.net/21.15107/rcub_cer_7345
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",
pages = "23-23",
url = "https://hdl.handle.net/21.15107/rcub_cer_7345"
}

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_cer_7345

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_cer_7345 .

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_cer_7345 .

TY  - CONF
AU  - Stefanović, Marija
AU  - Savić, Aleksa
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Radosavljević, Jelena
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7346
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
T1  - Poster presentation: Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains
UR  - https://hdl.handle.net/21.15107/rcub_cer_7346
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",
title = "Poster presentation: Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains",
url = "https://hdl.handle.net/21.15107/rcub_cer_7346"
}

Stefanović, M., Savić, A., Božić, N., Vujčić, Z.,& Radosavljević, J.. (2023). Poster presentation: Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains. 
Prirodno-matematički fakultet, Univerzitet u Kragujevcu..
https://hdl.handle.net/21.15107/rcub_cer_7346

Stefanović M, Savić A, Božić N, Vujčić Z, Radosavljević J. Poster presentation: Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains. 2023;.
https://hdl.handle.net/21.15107/rcub_cer_7346 .

Stefanović, Marija, Savić, Aleksa, Božić, Nataša, Vujčić, Zoran, Radosavljević, Jelena, "Poster presentation: Electrophoretic assessment of recombinant λ- exonuclease production in different E. coli strains" (2023),
https://hdl.handle.net/21.15107/rcub_cer_7346 .

TY  - CONF
AU  - Stojanović, Sanja
AU  - Stepanović, Jelena
AU  - Ristović, Marina
AU  - Dojnov, Biljana
AU  - Božić, Nataša
AU  - Duduk, Bojan
AU  - Vujčić, Zoran
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/5920
AB  - Fungal inulinases have wide application in industrial biotechnology, and it is presumed that their expression is regulated at the transcriptional level via promoter. It is also known that different sugars have an inducing effect on gene expression in fungal genome, including inulinases. Aim of this work was to determine which of the sugars used in growth medium, as the only carbon source, induce the extracellular exoinulinase gene inuE expression in Aspergillus welwitschiae FAW1. Inulin, rafinose, sucrose, glucose and fructose were used as carbon sources, and expression of inuE was monitored during 72 h of cultivation (tested after 24, 36, 48 and 72 h). Both, presence of mRNA in the mycelia and extracellular enzyme activity in the growth media were monitored. Interestingly,
obtained results showed that inuE was induced by fructose, sucrose and rafinose and not by inulin. In all cases, the highest mRNA was detected after 24 h of cultivation, while extracellular exoinulinase activity increased from 24 h with a peak in 72 h. Further experiments are necessary for a comprehensive understanding of the regulation mechanisms of AweinuE promoter for its more purposeful application in biotechnology.
PB  - University of Belgrade - Faculty of Chemistry
PB  - Serbian Biochemical Society
C3  - Proceedings - XI Conference of Serbian Biochemical Society "Amazing Biochemistry", 22.09.2022. Novi Sad, Serbia, 2022
T1  - Exoinulinase gene expression in Aspergillus welwitschiae FAW1 induced by different carbon sources
SP  - 141
UR  - https://hdl.handle.net/21.15107/rcub_cer_5920
ER  - 

@conference{
author = "Stojanović, Sanja and Stepanović, Jelena and Ristović, Marina and Dojnov, Biljana and Božić, Nataša and Duduk, Bojan and Vujčić, Zoran",
year = "2022",
abstract = "Fungal inulinases have wide application in industrial biotechnology, and it is presumed that their expression is regulated at the transcriptional level via promoter. It is also known that different sugars have an inducing effect on gene expression in fungal genome, including inulinases. Aim of this work was to determine which of the sugars used in growth medium, as the only carbon source, induce the extracellular exoinulinase gene inuE expression in Aspergillus welwitschiae FAW1. Inulin, rafinose, sucrose, glucose and fructose were used as carbon sources, and expression of inuE was monitored during 72 h of cultivation (tested after 24, 36, 48 and 72 h). Both, presence of mRNA in the mycelia and extracellular enzyme activity in the growth media were monitored. Interestingly,
obtained results showed that inuE was induced by fructose, sucrose and rafinose and not by inulin. In all cases, the highest mRNA was detected after 24 h of cultivation, while extracellular exoinulinase activity increased from 24 h with a peak in 72 h. Further experiments are necessary for a comprehensive understanding of the regulation mechanisms of AweinuE promoter for its more purposeful application in biotechnology.",
publisher = "University of Belgrade - Faculty of Chemistry, Serbian Biochemical Society",
journal = "Proceedings - XI Conference of Serbian Biochemical Society "Amazing Biochemistry", 22.09.2022. Novi Sad, Serbia, 2022",
title = "Exoinulinase gene expression in Aspergillus welwitschiae FAW1 induced by different carbon sources",
pages = "141",
url = "https://hdl.handle.net/21.15107/rcub_cer_5920"
}

Stojanović, S., Stepanović, J., Ristović, M., Dojnov, B., Božić, N., Duduk, B.,& Vujčić, Z.. (2022). Exoinulinase gene expression in Aspergillus welwitschiae FAW1 induced by different carbon sources. in Proceedings - XI Conference of Serbian Biochemical Society "Amazing Biochemistry", 22.09.2022. Novi Sad, Serbia, 2022
University of Belgrade - Faculty of Chemistry., 141.
https://hdl.handle.net/21.15107/rcub_cer_5920

Stojanović S, Stepanović J, Ristović M, Dojnov B, Božić N, Duduk B, Vujčić Z. Exoinulinase gene expression in Aspergillus welwitschiae FAW1 induced by different carbon sources. in Proceedings - XI Conference of Serbian Biochemical Society "Amazing Biochemistry", 22.09.2022. Novi Sad, Serbia, 2022. 2022;:141.
https://hdl.handle.net/21.15107/rcub_cer_5920 .

Stojanović, Sanja, Stepanović, Jelena, Ristović, Marina, Dojnov, Biljana, Božić, Nataša, Duduk, Bojan, Vujčić, Zoran, "Exoinulinase gene expression in Aspergillus welwitschiae FAW1 induced by different carbon sources" in Proceedings - XI Conference of Serbian Biochemical Society "Amazing Biochemistry", 22.09.2022. Novi Sad, Serbia, 2022 (2022):141,
https://hdl.handle.net/21.15107/rcub_cer_5920 .

TY  - CONF
AU  - Kosić, Višnja
AU  - Pavlović, Stefan
AU  - Božić, Nataša
AU  - Dojnov, Biljana
AU  - Stevanović, Gordana
AU  - Knežević-Jugović, Zorica
AU  - Milutinović Nikolić, Aleksandra
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/5925
AB  - α-Amylase from Bacillus paralicheniformis (BliAmy) is a highly efficient raw starch digesting enzyme. Starch is an inexpensive source of many food industrial products. Naturally occurring clay are non-toxic, environmentally friendly and inexpensive. Therefore, immobilization of BliAmy by adsorption on three differently modified bentonites was studied. Modifications included common Na-exchange procedure, acid activation, and alkali activation. The modified clays were characterized by X-ray powder diffraction, mercury intrusion porosimetry and the points of zero charge were determined. The adsorption of the enzyme was significantly influenced by the type of modification of bentonite, being the highest for the acid-activated bentonite with the highest porosity. On the other hand, the highest enzyme activity for immobilized α-amylase was obtained with alkali-modified bentonite (98 U/g), suggesting it as a good candidate for immobilization of α-amylase for application in the food industry.
PB  - The Society of Physical Chemists of Serbia
C3  - Proceedings - 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry, Physical Chemistry 2022, September, 26-30, 2022, Belgrade, Serbia
T1  - Immobilization of α-amylase from bacillus paralicheniformis on bentonites
SP  - 161
EP  - 164
UR  - https://hdl.handle.net/21.15107/rcub_cer_5925
ER  - 

@conference{
author = "Kosić, Višnja and Pavlović, Stefan and Božić, Nataša and Dojnov, Biljana and Stevanović, Gordana and Knežević-Jugović, Zorica and Milutinović Nikolić, Aleksandra",
year = "2022",
abstract = "α-Amylase from Bacillus paralicheniformis (BliAmy) is a highly efficient raw starch digesting enzyme. Starch is an inexpensive source of many food industrial products. Naturally occurring clay are non-toxic, environmentally friendly and inexpensive. Therefore, immobilization of BliAmy by adsorption on three differently modified bentonites was studied. Modifications included common Na-exchange procedure, acid activation, and alkali activation. The modified clays were characterized by X-ray powder diffraction, mercury intrusion porosimetry and the points of zero charge were determined. The adsorption of the enzyme was significantly influenced by the type of modification of bentonite, being the highest for the acid-activated bentonite with the highest porosity. On the other hand, the highest enzyme activity for immobilized α-amylase was obtained with alkali-modified bentonite (98 U/g), suggesting it as a good candidate for immobilization of α-amylase for application in the food industry.",
publisher = "The Society of Physical Chemists of Serbia",
journal = "Proceedings - 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry, Physical Chemistry 2022, September, 26-30, 2022, Belgrade, Serbia",
title = "Immobilization of α-amylase from bacillus paralicheniformis on bentonites",
pages = "161-164",
url = "https://hdl.handle.net/21.15107/rcub_cer_5925"
}

Kosić, V., Pavlović, S., Božić, N., Dojnov, B., Stevanović, G., Knežević-Jugović, Z.,& Milutinović Nikolić, A.. (2022). Immobilization of α-amylase from bacillus paralicheniformis on bentonites. in Proceedings - 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry, Physical Chemistry 2022, September, 26-30, 2022, Belgrade, Serbia
The Society of Physical Chemists of Serbia., 161-164.
https://hdl.handle.net/21.15107/rcub_cer_5925

Kosić V, Pavlović S, Božić N, Dojnov B, Stevanović G, Knežević-Jugović Z, Milutinović Nikolić A. Immobilization of α-amylase from bacillus paralicheniformis on bentonites. in Proceedings - 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry, Physical Chemistry 2022, September, 26-30, 2022, Belgrade, Serbia. 2022;:161-164.
https://hdl.handle.net/21.15107/rcub_cer_5925 .

Kosić, Višnja, Pavlović, Stefan, Božić, Nataša, Dojnov, Biljana, Stevanović, Gordana, Knežević-Jugović, Zorica, Milutinović Nikolić, Aleksandra, "Immobilization of α-amylase from bacillus paralicheniformis on bentonites" in Proceedings - 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry, Physical Chemistry 2022, September, 26-30, 2022, Belgrade, Serbia (2022):161-164,
https://hdl.handle.net/21.15107/rcub_cer_5925 .

TY  - CONF
AU  - Pavlović, Marija
AU  - Stojanović, Sanja
AU  - Dojnov, Biljana
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Margetić, Aleksandra
PY  - 2021
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/5834
AB  - Pectinolytic enzymes represent a large group of enzymes that catalyze the reactions of depolymerization and deesterification of pectin polysaccharides1. Saprophytic fungi produce pectinases on a large scale for industrial purposes. These enzymes have a various biotechnological application and their global annual production represents 25% of total industrial enzymes 1,2. Agro-waste is widely used as economical substrate for the production of pectinases by solid state fermentation 3. In this study, sugar beet pulp, as a good source of pectin3, was used as a substrate for enzyme production by Aspergillus tubingensis. This strain was isolated from the quince fruit and identified by the molecular DNA marker calmodulin (CaM). SSF was performed with this strain on sugar beet pulp (80%) in combination with wheat bran (20%), a potent substrate for pectinase production 3. The obtained high pectinolytic activity (15 U/mL), determined by 3,5-dinitrosalicylic acid reagent, was in the range of commercial pectinases. Zymography detection, using Ruthenium Red to visualize endo-pectinase activity and pectin-methyl esterase activity revealed several pectinase activity bands. Hydrolysis of different pectin substrates with the obtained pectinase complex was analyzed by thin layer chromatography in order to detect different products such as pectic oligosaccharides, which are emerging prebiotics superior to intact pectin.
PB  - University of Belgrade - Faculty of Chemistry
PB  - Serbian Biochemical Society
C3  - Proceedings - X Conference of Serbian Biochemical Society with international participation, “Biochemical Insights into Molecular Mechanisms”, 24.09.2021. Kragujevac, Serbia
T1  - Highly active pectinases from newly isolated Aspergillus tubingensis strain
SP  - 124
EP  - 125
UR  - https://hdl.handle.net/21.15107/rcub_cer_5834
ER  - 

@conference{
author = "Pavlović, Marija and Stojanović, Sanja and Dojnov, Biljana and Božić, Nataša and Vujčić, Zoran and Margetić, Aleksandra",
year = "2021",
abstract = "Pectinolytic enzymes represent a large group of enzymes that catalyze the reactions of depolymerization and deesterification of pectin polysaccharides1. Saprophytic fungi produce pectinases on a large scale for industrial purposes. These enzymes have a various biotechnological application and their global annual production represents 25% of total industrial enzymes 1,2. Agro-waste is widely used as economical substrate for the production of pectinases by solid state fermentation 3. In this study, sugar beet pulp, as a good source of pectin3, was used as a substrate for enzyme production by Aspergillus tubingensis. This strain was isolated from the quince fruit and identified by the molecular DNA marker calmodulin (CaM). SSF was performed with this strain on sugar beet pulp (80%) in combination with wheat bran (20%), a potent substrate for pectinase production 3. The obtained high pectinolytic activity (15 U/mL), determined by 3,5-dinitrosalicylic acid reagent, was in the range of commercial pectinases. Zymography detection, using Ruthenium Red to visualize endo-pectinase activity and pectin-methyl esterase activity revealed several pectinase activity bands. Hydrolysis of different pectin substrates with the obtained pectinase complex was analyzed by thin layer chromatography in order to detect different products such as pectic oligosaccharides, which are emerging prebiotics superior to intact pectin.",
publisher = "University of Belgrade - Faculty of Chemistry, Serbian Biochemical Society",
journal = "Proceedings - X Conference of Serbian Biochemical Society with international participation, “Biochemical Insights into Molecular Mechanisms”, 24.09.2021. Kragujevac, Serbia",
title = "Highly active pectinases from newly isolated Aspergillus tubingensis strain",
pages = "124-125",
url = "https://hdl.handle.net/21.15107/rcub_cer_5834"
}

Pavlović, M., Stojanović, S., Dojnov, B., Božić, N., Vujčić, Z.,& Margetić, A.. (2021). Highly active pectinases from newly isolated Aspergillus tubingensis strain. in Proceedings - X Conference of Serbian Biochemical Society with international participation, “Biochemical Insights into Molecular Mechanisms”, 24.09.2021. Kragujevac, Serbia
University of Belgrade - Faculty of Chemistry., 124-125.
https://hdl.handle.net/21.15107/rcub_cer_5834

Pavlović M, Stojanović S, Dojnov B, Božić N, Vujčić Z, Margetić A. Highly active pectinases from newly isolated Aspergillus tubingensis strain. in Proceedings - X Conference of Serbian Biochemical Society with international participation, “Biochemical Insights into Molecular Mechanisms”, 24.09.2021. Kragujevac, Serbia. 2021;:124-125.
https://hdl.handle.net/21.15107/rcub_cer_5834 .

Pavlović, Marija, Stojanović, Sanja, Dojnov, Biljana, Božić, Nataša, Vujčić, Zoran, Margetić, Aleksandra, "Highly active pectinases from newly isolated Aspergillus tubingensis strain" in Proceedings - X Conference of Serbian Biochemical Society with international participation, “Biochemical Insights into Molecular Mechanisms”, 24.09.2021. Kragujevac, Serbia (2021):124-125,
https://hdl.handle.net/21.15107/rcub_cer_5834 .

TY  - CONF
AU  - Božić, Nataša
PY  - 2021
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/5860
AB  - Understanding the structural peculiarities and properties of both, raw starch as a substrate and raw starch digesting amylases (RSDA) as an enzymes is needed for efficient application in food and other industries. Although knowledge of these structures and properties has increased significantly in recent years, the main requirement defining whether RSDA will be efficient in raw starch hydrolysis is still a riddle. We have recently identified the surface binding site (SBS) of a potent RSDA from Bacillus paralicheniformis ATCC 9945a (BliAmy), by crystallographic study of its native form and in complexes with maltose, acarbose, maltohexaose and β-cyclodextrin. To understand role of this SBS, the two key residues identified, Phe257 and Tyr358, were mutated. Kinetic studies show that starch binding through the SBS is disrupted in the mutants and that both mutants contributed cumulatively to binding and degradation. Mutation of both sites resulted in at least 5.5 fold weaker binding and 5 fold lower efficacy with raw starch as substrate 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 granule.
RSDA was further exploited for its robustness by changing its activity and converting this hydrolase into transglycosylase. The use of transglycosylases for synthetic purposes is limited since, unlike hydrolases, these enzymes are relatively rare in nature, and act on a limited substrate repertoire. To alter the activity of BliAmy, His235 was replaced with Glu. The mode of action of the mutant enzyme was tested using substrates such as starch, amylopectin, maltooligosaccharides etc. Mutant exhibited transglycosylation activity, while wild type BliAmy exhibited hydrolysis activity exclusively. Converting hyperthermostable BliAmy into transglycosylase provides potent tool in the synthesis of starch derivatives. The production of starchy foods with slow digestion properties, and thus a low glycemic index, is therefore an important goal of the modern food industry.
PB  - University of Belgrade
C3  - Book of Abstracts - 2nd UNIFood International Conference - UNIFood2021, September 24-25, 2021, Belgrade
T1  - From raw starch degrading α-amylase to transglycosylase by single point mutation
SP  - 13
EP  - 13
UR  - https://hdl.handle.net/21.15107/rcub_cer_5860
ER  - 

@conference{
author = "Božić, Nataša",
year = "2021",
abstract = "Understanding the structural peculiarities and properties of both, raw starch as a substrate and raw starch digesting amylases (RSDA) as an enzymes is needed for efficient application in food and other industries. Although knowledge of these structures and properties has increased significantly in recent years, the main requirement defining whether RSDA will be efficient in raw starch hydrolysis is still a riddle. We have recently identified the surface binding site (SBS) of a potent RSDA from Bacillus paralicheniformis ATCC 9945a (BliAmy), by crystallographic study of its native form and in complexes with maltose, acarbose, maltohexaose and β-cyclodextrin. To understand role of this SBS, the two key residues identified, Phe257 and Tyr358, were mutated. Kinetic studies show that starch binding through the SBS is disrupted in the mutants and that both mutants contributed cumulatively to binding and degradation. Mutation of both sites resulted in at least 5.5 fold weaker binding and 5 fold lower efficacy with raw starch as substrate 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 granule.
RSDA was further exploited for its robustness by changing its activity and converting this hydrolase into transglycosylase. The use of transglycosylases for synthetic purposes is limited since, unlike hydrolases, these enzymes are relatively rare in nature, and act on a limited substrate repertoire. To alter the activity of BliAmy, His235 was replaced with Glu. The mode of action of the mutant enzyme was tested using substrates such as starch, amylopectin, maltooligosaccharides etc. Mutant exhibited transglycosylation activity, while wild type BliAmy exhibited hydrolysis activity exclusively. Converting hyperthermostable BliAmy into transglycosylase provides potent tool in the synthesis of starch derivatives. The production of starchy foods with slow digestion properties, and thus a low glycemic index, is therefore an important goal of the modern food industry.",
publisher = "University of Belgrade",
journal = "Book of Abstracts - 2nd UNIFood International Conference - UNIFood2021, September 24-25, 2021, Belgrade",
title = "From raw starch degrading α-amylase to transglycosylase by single point mutation",
pages = "13-13",
url = "https://hdl.handle.net/21.15107/rcub_cer_5860"
}

Božić, N.. (2021). From raw starch degrading α-amylase to transglycosylase by single point mutation. in Book of Abstracts - 2nd UNIFood International Conference - UNIFood2021, September 24-25, 2021, Belgrade
University of Belgrade., 13-13.
https://hdl.handle.net/21.15107/rcub_cer_5860

Božić N. From raw starch degrading α-amylase to transglycosylase by single point mutation. in Book of Abstracts - 2nd UNIFood International Conference - UNIFood2021, September 24-25, 2021, Belgrade. 2021;:13-13.
https://hdl.handle.net/21.15107/rcub_cer_5860 .

Božić, Nataša, "From raw starch degrading α-amylase to transglycosylase by single point mutation" in Book of Abstracts - 2nd UNIFood International Conference - UNIFood2021, September 24-25, 2021, Belgrade (2021):13-13,
https://hdl.handle.net/21.15107/rcub_cer_5860 .

TY  - JOUR
AU  - Simić, Stefan
AU  - Jeremić, Sanja
AU  - Đokić, Lidija
AU  - Božić, Nataša
AU  - Vujčić, Zoran
AU  - Lončar, Nikola
AU  - Senthamaraikannan, Ramsankar
AU  - Babu, Ramesh Padamati
AU  - Opsenica, Igor
AU  - Nikodinović-Runić, Jasmina
PY  - 2020
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/3052
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.
PB  - Elsevier
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
SP  - 109411
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 and Senthamaraikannan, Ramsankar and Babu, Ramesh Padamati 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.",
publisher = "Elsevier",
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",
pages = "109411",
doi = "10.1016/j.enzmictec.2019.109411"
}

Simić, S., Jeremić, S., Đokić, L., Božić, N., Vujčić, Z., Lončar, N., 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
Elsevier., 132, 109411.
https://doi.org/10.1016/j.enzmictec.2019.109411

Simić S, Jeremić S, Đokić L, Božić N, Vujčić Z, Lončar N, 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;132:109411.
doi:10.1016/j.enzmictec.2019.109411 .

Simić, Stefan, Jeremić, Sanja, Đokić, Lidija, Božić, Nataša, Vujčić, Zoran, Lončar, Nikola, Senthamaraikannan, Ramsankar, Babu, Ramesh Padamati, 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):109411,
https://doi.org/10.1016/j.enzmictec.2019.109411 . .

TY  - JOUR
AU  - Božić, Nataša
AU  - Rozeboom, Henriëtte J.
AU  - Lončar, Nikola
AU  - Šokarda Slavić, Marinela
AU  - Janssen, Dick B.
AU  - Vujčić, Zoran
PY  - 2020
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/3728
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 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., 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 N, 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, 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
AU  - Šokarda Slavić, Marinela
AU  - Janssen, Dick B.
AU  - Vujčić, Zoran
PY  - 2020
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/3736
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 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., 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 N, 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, 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
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://cer.ihtm.bg.ac.rs/handle/123456789/3021
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 e ciency of dyeing processes
is still poor and results in large amounts of colored e uents. 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 (PfDyP B2) could be overexpressed as a soluble protein. PfDyP 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 PfDyP B2 in calcium-alginate beads resulted in a significant increase in stability: PfDyP B2 retains 80% of its initial activity after 2 h incubation at 50 °C, while the soluble enzyme is inactivated within minutes. PfDyP 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
SP  - 463
DO  - 10.3390/catal9050463
ER  - 

@article{
author = "Lončar, Nikola 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 e ciency of dyeing processes
is still poor and results in large amounts of colored e uents. 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 (PfDyP B2) could be overexpressed as a soluble protein. PfDyP 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 PfDyP B2 in calcium-alginate beads resulted in a significant increase in stability: PfDyP B2 retains 80% of its initial activity after 2 h incubation at 50 °C, while the soluble enzyme is inactivated within minutes. PfDyP 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",
pages = "463",
doi = "10.3390/catal9050463"
}

Lončar, N., 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, 463.
https://doi.org/10.3390/catal9050463

Lončar N, 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;9:463.
doi:10.3390/catal9050463 .

Lončar, Nikola, 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 (2019):463,
https://doi.org/10.3390/catal9050463 . .

TY  - CONF
AU  - Simić, Stefan
AU  - Božić, Nataša
AU  - Đokić, Lidija
AU  - Nikodinović-Runić, Jasmina
AU  - Opsenica, Igor
PY  - 2019
UR  - https://www.shd.org.rs/index.php/abstracts-56
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/3302
AB  - Laccases are a versatile class of enzymes with applications ranging from waste valorization to organic synthesis. We have tested whole-cell systems containing bacterial laccase as catalysts in the oxidation of 1,4-dihydropyridines. E. coli was used as the expression host for the cotA gene from Bacillus licheniformis, and the resulting whole-cell catalyst facilitated the oxidation of 1,4-dihydropyridines. It was found that multicopper oxidase CueO from the E. coli expression host also possesses catalytic activity in the oxidation of 1,4-dihydropyridines. The whole-cell biocatalyst expressing Bacillus licheniformis laccase was subsequently immobilized on bacterial nanocellulose and utilized in the same transformation, retaining 37 % of its original activity after three consecutive catalytic runs. This is the first report of a whole-cell catalytic system containing recombinant laccase for the oxidation of 1,4-dihydropyridines.
AB  - Lakaze predstavljalju raznoliku klasu enzima koja nalazi primenu od valorizacije otpada do organske sinteze. U ovom istraživanju ispitivane su cele ćelije koje sadrže bakterijsku lakazu kao katalizator u oksidaciji 1,4-dihidropiridina. Ekspresija cotA gena iz Bacillus licheniformis je izvršena u ćelijama E. coli i za nastali biokatalizator je ustanovljeno da ubrzava oksidaciju 1,4-dihidropiridina. Pored toga, ustanovljeno je da „multicopper“ oksidaza CueO iz E. coli takođe poseduje aktivnost prema oksidaciji 1,4-dihidropiridina. Ekspresioni sistem koji sadrži lakazu iz bakterije Bacillus licheniformis zatim je imobilizovan na bakterijskoj nanocelulozi i upotrebljen je kao katalizator u istoj transformaciji. Takav katalizator je bilo moguće ponovo upotrebiti tri puta, nakon čega je njegova aktivnost iznosila 37 % od početne. Navedeno istraživanje predstavlja prvu primenu celih ćelija sa rekombinantnom lakazom u oksidaciji 1,4-dihidropiridina.
PB  - Serbian Chemical Society, Belgrade / Srpsko hemijsko društvo, Beograd
C3  - 56th Meeting of the Serbian chemical Society - Book of Abstracts / 56. Savetovanje Srpskog hemijskog društva - Kratki izvodi radova, Niš 7-8.9. 2019.
T1  - Oxidation of 1,4-dihydropyridines catalyzed by recombinant bacterial laccase expressed in E. coli
T1  - Oksidacija 1,4-dihidropiridina katalizovana rekombinantnom bakterijskom lakazom eksprimiranom u E. coli
SP  - 88
UR  - https://hdl.handle.net/21.15107/rcub_cer_3302
ER  - 

@conference{
author = "Simić, Stefan and Božić, Nataša and Đokić, Lidija and Nikodinović-Runić, Jasmina and Opsenica, Igor",
year = "2019",
abstract = "Laccases are a versatile class of enzymes with applications ranging from waste valorization to organic synthesis. We have tested whole-cell systems containing bacterial laccase as catalysts in the oxidation of 1,4-dihydropyridines. E. coli was used as the expression host for the cotA gene from Bacillus licheniformis, and the resulting whole-cell catalyst facilitated the oxidation of 1,4-dihydropyridines. It was found that multicopper oxidase CueO from the E. coli expression host also possesses catalytic activity in the oxidation of 1,4-dihydropyridines. The whole-cell biocatalyst expressing Bacillus licheniformis laccase was subsequently immobilized on bacterial nanocellulose and utilized in the same transformation, retaining 37 % of its original activity after three consecutive catalytic runs. This is the first report of a whole-cell catalytic system containing recombinant laccase for the oxidation of 1,4-dihydropyridines., Lakaze predstavljalju raznoliku klasu enzima koja nalazi primenu od valorizacije otpada do organske sinteze. U ovom istraživanju ispitivane su cele ćelije koje sadrže bakterijsku lakazu kao katalizator u oksidaciji 1,4-dihidropiridina. Ekspresija cotA gena iz Bacillus licheniformis je izvršena u ćelijama E. coli i za nastali biokatalizator je ustanovljeno da ubrzava oksidaciju 1,4-dihidropiridina. Pored toga, ustanovljeno je da „multicopper“ oksidaza CueO iz E. coli takođe poseduje aktivnost prema oksidaciji 1,4-dihidropiridina. Ekspresioni sistem koji sadrži lakazu iz bakterije Bacillus licheniformis zatim je imobilizovan na bakterijskoj nanocelulozi i upotrebljen je kao katalizator u istoj transformaciji. Takav katalizator je bilo moguće ponovo upotrebiti tri puta, nakon čega je njegova aktivnost iznosila 37 % od početne. Navedeno istraživanje predstavlja prvu primenu celih ćelija sa rekombinantnom lakazom u oksidaciji 1,4-dihidropiridina.",
publisher = "Serbian Chemical Society, Belgrade / Srpsko hemijsko društvo, Beograd",
journal = "56th Meeting of the Serbian chemical Society - Book of Abstracts / 56. Savetovanje Srpskog hemijskog društva - Kratki izvodi radova, Niš 7-8.9. 2019.",
title = "Oxidation of 1,4-dihydropyridines catalyzed by recombinant bacterial laccase expressed in E. coli, Oksidacija 1,4-dihidropiridina katalizovana rekombinantnom bakterijskom lakazom eksprimiranom u E. coli",
pages = "88",
url = "https://hdl.handle.net/21.15107/rcub_cer_3302"
}

Simić, S., Božić, N., Đokić, L., Nikodinović-Runić, J.,& Opsenica, I.. (2019). Oxidation of 1,4-dihydropyridines catalyzed by recombinant bacterial laccase expressed in E. coli. in 56th Meeting of the Serbian chemical Society - Book of Abstracts / 56. Savetovanje Srpskog hemijskog društva - Kratki izvodi radova, Niš 7-8.9. 2019.
Serbian Chemical Society, Belgrade / Srpsko hemijsko društvo, Beograd., 88.
https://hdl.handle.net/21.15107/rcub_cer_3302

Simić S, Božić N, Đokić L, Nikodinović-Runić J, Opsenica I. Oxidation of 1,4-dihydropyridines catalyzed by recombinant bacterial laccase expressed in E. coli. in 56th Meeting of the Serbian chemical Society - Book of Abstracts / 56. Savetovanje Srpskog hemijskog društva - Kratki izvodi radova, Niš 7-8.9. 2019.. 2019;:88.
https://hdl.handle.net/21.15107/rcub_cer_3302 .

Simić, Stefan, Božić, Nataša, Đokić, Lidija, Nikodinović-Runić, Jasmina, Opsenica, Igor, "Oxidation of 1,4-dihydropyridines catalyzed by recombinant bacterial laccase expressed in E. coli" in 56th Meeting of the Serbian chemical Society - Book of Abstracts / 56. Savetovanje Srpskog hemijskog društva - Kratki izvodi radova, Niš 7-8.9. 2019. (2019):88,
https://hdl.handle.net/21.15107/rcub_cer_3302 .

TY  - JOUR
AU  - Lončar, Nikola
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2016
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/1908
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.
PB  - Elsevier
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 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.",
publisher = "Elsevier",
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., 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., 134, 390-395.
https://doi.org/10.1016/j.molcatb.2016.06.005

Lončar N, 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, 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  - Šokarda Slavić, Marinela
AU  - Pešić, Milja
AU  - Vujčić, Zoran
AU  - Božić, Nataša
PY  - 2016
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/1965
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
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2016
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/3131
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.
PB  - Elsevier
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 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.",
publisher = "Elsevier",
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., 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., 134, 390-395.
https://doi.org/10.1016/j.molcatb.2016.06.005

Lončar N, 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, 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  - CONF
AU  - Šokarda Slavić, Marinela
AU  - Lončar, Nikola
AU  - Janeček, Štefan
AU  - Vujčić, Zoran
AU  - Božić, Nataša
PY  - 2016
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/3551
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Sixth Conference, “Biochemistry and Interdisciplinarity: Transcending the Limits of Field”, Faculty of Chemistry, University of Belgrade, 18.11.2016. Belgrade, Serbia
T1  - Rational design of raw starch degrading α-amylase from Bacillus licheniformis 9945a for possible surface binding sites indetification
SP  - 159
EP  - 159
UR  - https://hdl.handle.net/21.15107/rcub_cer_3551
ER  - 

@conference{
author = "Šokarda Slavić, Marinela and Lončar, Nikola and Janeček, Štefan and Vujčić, Zoran and Božić, Nataša",
year = "2016",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Sixth Conference, “Biochemistry and Interdisciplinarity: Transcending the Limits of Field”, Faculty of Chemistry, University of Belgrade, 18.11.2016. Belgrade, Serbia",
title = "Rational design of raw starch degrading α-amylase from Bacillus licheniformis 9945a for possible surface binding sites indetification",
pages = "159-159",
url = "https://hdl.handle.net/21.15107/rcub_cer_3551"
}

Šokarda Slavić, M., Lončar, N., Janeček, Š., Vujčić, Z.,& Božić, N.. (2016). Rational design of raw starch degrading α-amylase from Bacillus licheniformis 9945a for possible surface binding sites indetification. in Serbian Biochemical Society Sixth Conference, “Biochemistry and Interdisciplinarity: Transcending the Limits of Field”, Faculty of Chemistry, University of Belgrade, 18.11.2016. Belgrade, Serbia
Serbian Biochemical Society., 159-159.
https://hdl.handle.net/21.15107/rcub_cer_3551

Šokarda Slavić M, Lončar N, Janeček Š, Vujčić Z, Božić N. Rational design of raw starch degrading α-amylase from Bacillus licheniformis 9945a for possible surface binding sites indetification. in Serbian Biochemical Society Sixth Conference, “Biochemistry and Interdisciplinarity: Transcending the Limits of Field”, Faculty of Chemistry, University of Belgrade, 18.11.2016. Belgrade, Serbia. 2016;:159-159.
https://hdl.handle.net/21.15107/rcub_cer_3551 .

Šokarda Slavić, Marinela, Lončar, Nikola, Janeček, Štefan, Vujčić, Zoran, Božić, Nataša, "Rational design of raw starch degrading α-amylase from Bacillus licheniformis 9945a for possible surface binding sites indetification" in Serbian Biochemical Society Sixth Conference, “Biochemistry and Interdisciplinarity: Transcending the Limits of Field”, Faculty of Chemistry, University of Belgrade, 18.11.2016. Belgrade, Serbia (2016):159-159,
https://hdl.handle.net/21.15107/rcub_cer_3551 .

TY  - JOUR
AU  - Vujčić, Zoran
AU  - Janović, Barbara
AU  - Lončar, Nikola
AU  - Margetić, Aleksandra
AU  - Božić, Nataša
AU  - Dojnov, Biljana
AU  - Vujčić, Miroslava
PY  - 2015
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/1760
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.
PB  - Elsevier Sci Ltd, Oxford
T2  - International Biodeterioration & 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 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.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "International Biodeterioration & 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., Margetić, A., Božić, N., Dojnov, B.,& Vujčić, M.. (2015). Exploitation of neglected horseradish peroxidase izoenzymes for dye decolorization. in International Biodeterioration & Biodegradation
Elsevier Sci Ltd, Oxford., 97, 124-127.
https://doi.org/10.1016/j.ibiod.2014.10.007

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

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