TY  - JOUR
AU  - Andreu, Alicia
AU  - Ćorović, Marija
AU  - Garcia-Sanz, Carla
AU  - Santos, A. Sofia
AU  - Milivojević, Ana
AU  - Ortega-Nieto, Clara
AU  - Mateo, Cesar
AU  - Bezbradica, Dejan
AU  - Palomo, Jose M.
PY  - 2023
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/6804
AB  - Enzymatic glycosylation is a versatile and sustainable biotechnological approach that plays a pivotal role in the production of bioactive compounds. This process involves the enzymatic transfer of sugar moieties onto various acceptor molecules, such as small molecules, peptides, or proteins, resulting in the synthesis of glycosides. These glycosides often exhibit enhanced bioactivity, improved solubility, and enhanced stability, making them valuable in pharmaceuticals, nutraceuticals, and the food industry. This review explores the diverse enzymatic glycosylation strategies employed in the synthesis of bioactive compounds. It highlights the enzymatic catalysts involved, including glycosyltransferases, glycosidases, glycophosphorylases, and glycosynthases. It considers the advantages and disadvantages of these biocatalysts in the stereoselective and regioselective synthesis of different types of glycosylated molecules, phenolic and aliphatic alcohols, oligosaccharides, polysaccharides, glycoderivatives, glycopeptides, and glycoproteins with a clear focus on food and pharmaceutical chemistry. Furthermore, the review outlines various sources of sugar donors, activated glycosides, and sugar nucleotides, as well as the utilization of engineered enzymes and microorganisms for glycosylation reactions. The advantages of enzymatic glycosylation, including its high regioselectivity, stereoselectivity, and sustainability, are emphasized. Therefore, these approaches combining the use of different catalytic systems, the improvement of tools such as immobilization technology or chemical or genetic modification to improve the glycosylation process, could be useful tools in continuous biotechnological advancements.
PB  - MDPI
T2  - Catalysts
T1  - Enzymatic Glycosylation Strategies in the Production of Bioactive Compounds
IS  - 10
SP  - 1359
VL  - 13
DO  - 10.3390/catal13101359
ER  - 

@article{
author = "Andreu, Alicia and Ćorović, Marija and Garcia-Sanz, Carla and Santos, A. Sofia and Milivojević, Ana and Ortega-Nieto, Clara and Mateo, Cesar and Bezbradica, Dejan and Palomo, Jose M.",
year = "2023",
abstract = "Enzymatic glycosylation is a versatile and sustainable biotechnological approach that plays a pivotal role in the production of bioactive compounds. This process involves the enzymatic transfer of sugar moieties onto various acceptor molecules, such as small molecules, peptides, or proteins, resulting in the synthesis of glycosides. These glycosides often exhibit enhanced bioactivity, improved solubility, and enhanced stability, making them valuable in pharmaceuticals, nutraceuticals, and the food industry. This review explores the diverse enzymatic glycosylation strategies employed in the synthesis of bioactive compounds. It highlights the enzymatic catalysts involved, including glycosyltransferases, glycosidases, glycophosphorylases, and glycosynthases. It considers the advantages and disadvantages of these biocatalysts in the stereoselective and regioselective synthesis of different types of glycosylated molecules, phenolic and aliphatic alcohols, oligosaccharides, polysaccharides, glycoderivatives, glycopeptides, and glycoproteins with a clear focus on food and pharmaceutical chemistry. Furthermore, the review outlines various sources of sugar donors, activated glycosides, and sugar nucleotides, as well as the utilization of engineered enzymes and microorganisms for glycosylation reactions. The advantages of enzymatic glycosylation, including its high regioselectivity, stereoselectivity, and sustainability, are emphasized. Therefore, these approaches combining the use of different catalytic systems, the improvement of tools such as immobilization technology or chemical or genetic modification to improve the glycosylation process, could be useful tools in continuous biotechnological advancements.",
publisher = "MDPI",
journal = "Catalysts",
title = "Enzymatic Glycosylation Strategies in the Production of Bioactive Compounds",
number = "10",
pages = "1359",
volume = "13",
doi = "10.3390/catal13101359"
}

Andreu, A., Ćorović, M., Garcia-Sanz, C., Santos, A. S., Milivojević, A., Ortega-Nieto, C., Mateo, C., Bezbradica, D.,& Palomo, J. M.. (2023). Enzymatic Glycosylation Strategies in the Production of Bioactive Compounds. in Catalysts
MDPI., 13(10), 1359.
https://doi.org/10.3390/catal13101359

Andreu A, Ćorović M, Garcia-Sanz C, Santos AS, Milivojević A, Ortega-Nieto C, Mateo C, Bezbradica D, Palomo JM. Enzymatic Glycosylation Strategies in the Production of Bioactive Compounds. in Catalysts. 2023;13(10):1359.
doi:10.3390/catal13101359 .

Andreu, Alicia, Ćorović, Marija, Garcia-Sanz, Carla, Santos, A. Sofia, Milivojević, Ana, Ortega-Nieto, Clara, Mateo, Cesar, Bezbradica, Dejan, Palomo, Jose M., "Enzymatic Glycosylation Strategies in the Production of Bioactive Compounds" in Catalysts, 13, no. 10 (2023):1359,
https://doi.org/10.3390/catal13101359 . .

TY  - CONF
AU  - dos Santos, Leandro Alves
AU  - Banjanac, Katarina
AU  - Veljković, Milica
AU  - Simović, Milica
AU  - Mateo, Cesar
AU  - Bezbradica, Dejan
PY  - 2023
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/6948
AB  - A food-grade bacterial xylanase enzyme preparation, ROHALASE® SEP-VISCO,
was immobilized by adsorption onto polyethyleneimine (PEI) functionalized magnetic
nanoparticles (MNPs). This process resulted in nanobiocatalysts with optimal characteristics.
Subsequently, this nanobiocatalyst was employed to produce xylo-oligosaccharides (XOS). In
the past decade, magnetic nanoparticles (MNPs) have gained significant interest, finding
diverse applications in biomedicine, biosensor production, food processing, catalysis,
agriculture, and environmental processes. MNPs have proven highly effective for enzyme
immobilization because of their unique and distinguished properties such as high specific
surface area, biocompatibility, and magnetic characteristics. Magnetic characteristics
facilitate the easy removal of MNPs from reaction mixtures, allowing for their efficient reuse.
The functionalization of MNPs through with the cationic polymer PEI can further augment
their ability for enzyme attachment. Generally, xylanases hydrolyses β-1,4-glycosidic
linkages present in xylan which is a major component of the hemicellulosic fraction in the
plant cell walls. Furthermore, xylanases can produce XOS composed of xylose units linked
by β-1,4-xylosidic bonds. XOS have stimulatory effects on the selective growth of human
intestinal microbiota and are frequently defined as prebiotics.
In the present work, the xylanase immobilization efficiency on MNPs-PEI is between
100 and 43 % within the wide range of xylanase concentrations (400-2400 mg/g of support).
Free and immobilized xylanase showed maximal catalytic activity at pH 6.0 and 60°C in
reaction with commercial birchwood xylan (concentration of 1 % w/v). The maximum
activity of 1675 IU/g of support was achieved when immobilization was performed at initial
enzyme concentration of 1250 mg/g of support during 2 h. Since this immobilized
preparation exhibited the activity immobilization yield of 80 % and specific activity of 2.1
mg of proteins/g of support, it has been applied in reaction of XOS synthesis. The MNPsPEI-xylanase was found to produce high yield of XOS from birchwood xylan, indicating its
potential for utilization in feed and food formulations.
PB  - Belgrade : University, Faculty of Technology and Metallurgy
C3  - Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade
T1  - IMMOBILIZATION OF XYLANASE ON MAGNETIC NANOPARTICLES MODIFIED WITH POLYETHYLENEIMINE AND ITS APPLICATION IN XYLO-OLIGOSACCHARIDES SYNTHESIS
SP  - 34
UR  - https://hdl.handle.net/21.15107/rcub_technorep_6948
ER  - 

@conference{
author = "dos Santos, Leandro Alves and Banjanac, Katarina and Veljković, Milica and Simović, Milica and Mateo, Cesar and Bezbradica, Dejan",
year = "2023",
abstract = "A food-grade bacterial xylanase enzyme preparation, ROHALASE® SEP-VISCO,
was immobilized by adsorption onto polyethyleneimine (PEI) functionalized magnetic
nanoparticles (MNPs). This process resulted in nanobiocatalysts with optimal characteristics.
Subsequently, this nanobiocatalyst was employed to produce xylo-oligosaccharides (XOS). In
the past decade, magnetic nanoparticles (MNPs) have gained significant interest, finding
diverse applications in biomedicine, biosensor production, food processing, catalysis,
agriculture, and environmental processes. MNPs have proven highly effective for enzyme
immobilization because of their unique and distinguished properties such as high specific
surface area, biocompatibility, and magnetic characteristics. Magnetic characteristics
facilitate the easy removal of MNPs from reaction mixtures, allowing for their efficient reuse.
The functionalization of MNPs through with the cationic polymer PEI can further augment
their ability for enzyme attachment. Generally, xylanases hydrolyses β-1,4-glycosidic
linkages present in xylan which is a major component of the hemicellulosic fraction in the
plant cell walls. Furthermore, xylanases can produce XOS composed of xylose units linked
by β-1,4-xylosidic bonds. XOS have stimulatory effects on the selective growth of human
intestinal microbiota and are frequently defined as prebiotics.
In the present work, the xylanase immobilization efficiency on MNPs-PEI is between
100 and 43 % within the wide range of xylanase concentrations (400-2400 mg/g of support).
Free and immobilized xylanase showed maximal catalytic activity at pH 6.0 and 60°C in
reaction with commercial birchwood xylan (concentration of 1 % w/v). The maximum
activity of 1675 IU/g of support was achieved when immobilization was performed at initial
enzyme concentration of 1250 mg/g of support during 2 h. Since this immobilized
preparation exhibited the activity immobilization yield of 80 % and specific activity of 2.1
mg of proteins/g of support, it has been applied in reaction of XOS synthesis. The MNPsPEI-xylanase was found to produce high yield of XOS from birchwood xylan, indicating its
potential for utilization in feed and food formulations.",
publisher = "Belgrade : University, Faculty of Technology and Metallurgy",
journal = "Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade",
title = "IMMOBILIZATION OF XYLANASE ON MAGNETIC NANOPARTICLES MODIFIED WITH POLYETHYLENEIMINE AND ITS APPLICATION IN XYLO-OLIGOSACCHARIDES SYNTHESIS",
pages = "34",
url = "https://hdl.handle.net/21.15107/rcub_technorep_6948"
}

dos Santos, L. A., Banjanac, K., Veljković, M., Simović, M., Mateo, C.,& Bezbradica, D.. (2023). IMMOBILIZATION OF XYLANASE ON MAGNETIC NANOPARTICLES MODIFIED WITH POLYETHYLENEIMINE AND ITS APPLICATION IN XYLO-OLIGOSACCHARIDES SYNTHESIS. in Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade
Belgrade : University, Faculty of Technology and Metallurgy., 34.
https://hdl.handle.net/21.15107/rcub_technorep_6948

dos Santos LA, Banjanac K, Veljković M, Simović M, Mateo C, Bezbradica D. IMMOBILIZATION OF XYLANASE ON MAGNETIC NANOPARTICLES MODIFIED WITH POLYETHYLENEIMINE AND ITS APPLICATION IN XYLO-OLIGOSACCHARIDES SYNTHESIS. in Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade. 2023;:34.
https://hdl.handle.net/21.15107/rcub_technorep_6948 .

dos Santos, Leandro Alves, Banjanac, Katarina, Veljković, Milica, Simović, Milica, Mateo, Cesar, Bezbradica, Dejan, "IMMOBILIZATION OF XYLANASE ON MAGNETIC NANOPARTICLES MODIFIED WITH POLYETHYLENEIMINE AND ITS APPLICATION IN XYLO-OLIGOSACCHARIDES SYNTHESIS" in Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade (2023):34,
https://hdl.handle.net/21.15107/rcub_technorep_6948 .

TY  - JOUR
AU  - Bezbradica, Dejan
AU  - Mateo, Cesar
AU  - Guisan, Jose M.
PY  - 2014
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/2813
AB  - Immobilization of enzymes on glutaraldehyde-activated supports has been largely used on supports previously activated with amine groups. Therefore, the supports are positively charged hence usually the immobilization is promoted through a two step mechanism: in a first step the enzyme is adsorbed on the support via an anionic exchange mechanism and then, the covalent immobilization occurs. In this paper a new glutaraldehyde activated support without a net charge is presented and characterized in immobilizations of trypsin, penicillin acylase G, lipase and E. coli BL21 cell extract. Immobilization mechanism was studied and this was produced without an adsorption step. This support promoted initially a reversible immobilization, converting into irreversible after incubation of the enzyme-support for several days or after a reduction step. In addition the stability of glutaraldehyde groups was studied retaining around 50 and 25% of its immobilization capacity for 24 h at pH 7 and 10 respectively. This fact allows the incubation of the enzyme with the support even at alkaline pH promoting an extra stabilization factor for trypsin on this support.
PB  - Elsevier, Amsterdam
T2  - Journal of Molecular Catalysis B-Enzymatic
T1  - Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde
EP  - 224
SP  - 218
VL  - 102
DO  - 10.1016/j.molcatb.2014.02.021
ER  - 

@article{
author = "Bezbradica, Dejan and Mateo, Cesar and Guisan, Jose M.",
year = "2014",
abstract = "Immobilization of enzymes on glutaraldehyde-activated supports has been largely used on supports previously activated with amine groups. Therefore, the supports are positively charged hence usually the immobilization is promoted through a two step mechanism: in a first step the enzyme is adsorbed on the support via an anionic exchange mechanism and then, the covalent immobilization occurs. In this paper a new glutaraldehyde activated support without a net charge is presented and characterized in immobilizations of trypsin, penicillin acylase G, lipase and E. coli BL21 cell extract. Immobilization mechanism was studied and this was produced without an adsorption step. This support promoted initially a reversible immobilization, converting into irreversible after incubation of the enzyme-support for several days or after a reduction step. In addition the stability of glutaraldehyde groups was studied retaining around 50 and 25% of its immobilization capacity for 24 h at pH 7 and 10 respectively. This fact allows the incubation of the enzyme with the support even at alkaline pH promoting an extra stabilization factor for trypsin on this support.",
publisher = "Elsevier, Amsterdam",
journal = "Journal of Molecular Catalysis B-Enzymatic",
title = "Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde",
pages = "224-218",
volume = "102",
doi = "10.1016/j.molcatb.2014.02.021"
}

Bezbradica, D., Mateo, C.,& Guisan, J. M.. (2014). Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde. in Journal of Molecular Catalysis B-Enzymatic
Elsevier, Amsterdam., 102, 218-224.
https://doi.org/10.1016/j.molcatb.2014.02.021

Bezbradica D, Mateo C, Guisan JM. Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde. in Journal of Molecular Catalysis B-Enzymatic. 2014;102:218-224.
doi:10.1016/j.molcatb.2014.02.021 .

Bezbradica, Dejan, Mateo, Cesar, Guisan, Jose M., "Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde" in Journal of Molecular Catalysis B-Enzymatic, 102 (2014):218-224,
https://doi.org/10.1016/j.molcatb.2014.02.021 . .