TY  - JOUR
AU  - Carević, Milica
AU  - Vukašinović-Sekulić, Maja
AU  - Ćorović, Marija
AU  - Rogniaux, Helene
AU  - Ropartz, David
AU  - Veličković, Dušan
AU  - Bezbradica, Dejan
PY  - 2018
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/3934
AB  - beta-Galactosidase is an important industrial enzyme that catalyzes reaction of lactose hydrolysis and recently more interesting reaction of transgalactosylation, yielding a highly valuable group of prebiotic compounds named galacto-oligosaccharides (GOS). In this paper, parameters for achieving high yields of tailor-made GOS using crude beta-galactosidase obtained from Lactobacillus acidophilus ATCC 4356, probiotic bacteria regarded as safe for human consumption, were optimized. At the same time, detailed structural elucidation of obtained GOS was conducted, and it was concluded that beta-galactosidase from L acidophilus shows a particular specificity towards the formation of beta-(1 - gt  6) glycosidic bonds. In order to develop more stable and economically cost-effective preparation, crude enzyme was successfully immobilized on a methacrylic polymer carrier Lifetech ECR8409, leading to its simultaneous 2-fold purification. This immobilized preparation showed unchanged specificity towards the transgalactosylation reaction, thus yielding 86 WI GOS under the previously optimized conditions (lactose concentration 400 g/l in 0.1 M sodium phosphate buffer, pH 6.8 and temperature 50 degrees C).
PB  - Soc Bioscience Bioengineering Japan, Osaka
T2  - Journal of Bioscience and Bioengineering
T1  - Evaluation of beta-galactosidase from Lactobacillus acidophilus as biocatalyst for galacto-oligosaccharides synthesis: Product structural characterization and enzyme immobilization
EP  - 704
IS  - 6
SP  - 697
VL  - 126
DO  - 10.1016/j.jbiosc.2018.06.003
ER  - 

@article{
author = "Carević, Milica and Vukašinović-Sekulić, Maja and Ćorović, Marija and Rogniaux, Helene and Ropartz, David and Veličković, Dušan and Bezbradica, Dejan",
year = "2018",
abstract = "beta-Galactosidase is an important industrial enzyme that catalyzes reaction of lactose hydrolysis and recently more interesting reaction of transgalactosylation, yielding a highly valuable group of prebiotic compounds named galacto-oligosaccharides (GOS). In this paper, parameters for achieving high yields of tailor-made GOS using crude beta-galactosidase obtained from Lactobacillus acidophilus ATCC 4356, probiotic bacteria regarded as safe for human consumption, were optimized. At the same time, detailed structural elucidation of obtained GOS was conducted, and it was concluded that beta-galactosidase from L acidophilus shows a particular specificity towards the formation of beta-(1 - gt  6) glycosidic bonds. In order to develop more stable and economically cost-effective preparation, crude enzyme was successfully immobilized on a methacrylic polymer carrier Lifetech ECR8409, leading to its simultaneous 2-fold purification. This immobilized preparation showed unchanged specificity towards the transgalactosylation reaction, thus yielding 86 WI GOS under the previously optimized conditions (lactose concentration 400 g/l in 0.1 M sodium phosphate buffer, pH 6.8 and temperature 50 degrees C).",
publisher = "Soc Bioscience Bioengineering Japan, Osaka",
journal = "Journal of Bioscience and Bioengineering",
title = "Evaluation of beta-galactosidase from Lactobacillus acidophilus as biocatalyst for galacto-oligosaccharides synthesis: Product structural characterization and enzyme immobilization",
pages = "704-697",
number = "6",
volume = "126",
doi = "10.1016/j.jbiosc.2018.06.003"
}

Carević, M., Vukašinović-Sekulić, M., Ćorović, M., Rogniaux, H., Ropartz, D., Veličković, D.,& Bezbradica, D.. (2018). Evaluation of beta-galactosidase from Lactobacillus acidophilus as biocatalyst for galacto-oligosaccharides synthesis: Product structural characterization and enzyme immobilization. in Journal of Bioscience and Bioengineering
Soc Bioscience Bioengineering Japan, Osaka., 126(6), 697-704.
https://doi.org/10.1016/j.jbiosc.2018.06.003

Carević M, Vukašinović-Sekulić M, Ćorović M, Rogniaux H, Ropartz D, Veličković D, Bezbradica D. Evaluation of beta-galactosidase from Lactobacillus acidophilus as biocatalyst for galacto-oligosaccharides synthesis: Product structural characterization and enzyme immobilization. in Journal of Bioscience and Bioengineering. 2018;126(6):697-704.
doi:10.1016/j.jbiosc.2018.06.003 .

Carević, Milica, Vukašinović-Sekulić, Maja, Ćorović, Marija, Rogniaux, Helene, Ropartz, David, Veličković, Dušan, Bezbradica, Dejan, "Evaluation of beta-galactosidase from Lactobacillus acidophilus as biocatalyst for galacto-oligosaccharides synthesis: Product structural characterization and enzyme immobilization" in Journal of Bioscience and Bioengineering, 126, no. 6 (2018):697-704,
https://doi.org/10.1016/j.jbiosc.2018.06.003 . .

TY  - JOUR
AU  - Simović, Milica
AU  - Bezbradica, Dejan
AU  - Banjanac, Katarina
AU  - Milivojević, Ana
AU  - Fanuel, Mathieu
AU  - Rogniaux, Helene
AU  - Ropartz, David
AU  - Veličković, Dušan
PY  - 2016
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/3258
AB  - Galacto-oligosaccharides (GOS) represent a diverse group of well-characterized prebiotic ingredients derived from lactose in a reaction catalyzed with beta-galactosidases. Enzymatic transgalactosylation results in a mixture of compounds of various degrees of polymerization and types of linkages. Because structure plays an important role in terms of prebiotic activity, it is of crucial importance to provide an insight into the mechanism of transgalactosylation reaction and occurrence of different types of beta-linkages during GOS synthesis. Our study proved that a novel one-step method, based on ion-mobility spectrometry tandem mass spectrometry (IMS-MS/MS), enables complete elucidation of GOS structure. It has been shown that) beta-galactosidase from Aspergillus oryzae has the highest affinity toward formation of beta-(1 - gt  3) or beta-(1 - gt  6) linkages. Additionally, it was observed that the occurrence of different linkages varies during the reaction course, indicating that tailoring favorable GOS structures with improved prebiotic activity can be achieved by adequate control of enzymatic synthesis.
PB  - Amer Chemical Soc, Washington
T2  - Journal of Agricultural and Food Chemistry
T1  - Structural Elucidation of Enzymatically Synthesized Galacto-oligosaccharides Using Ion-Mobility Spectrometry Tandem Mass Spectrometry
EP  - 3615
IS  - 18
SP  - 3609
VL  - 64
DO  - 10.1021/acs.jafc.6b01293
ER  - 

@article{
author = "Simović, Milica and Bezbradica, Dejan and Banjanac, Katarina and Milivojević, Ana and Fanuel, Mathieu and Rogniaux, Helene and Ropartz, David and Veličković, Dušan",
year = "2016",
abstract = "Galacto-oligosaccharides (GOS) represent a diverse group of well-characterized prebiotic ingredients derived from lactose in a reaction catalyzed with beta-galactosidases. Enzymatic transgalactosylation results in a mixture of compounds of various degrees of polymerization and types of linkages. Because structure plays an important role in terms of prebiotic activity, it is of crucial importance to provide an insight into the mechanism of transgalactosylation reaction and occurrence of different types of beta-linkages during GOS synthesis. Our study proved that a novel one-step method, based on ion-mobility spectrometry tandem mass spectrometry (IMS-MS/MS), enables complete elucidation of GOS structure. It has been shown that) beta-galactosidase from Aspergillus oryzae has the highest affinity toward formation of beta-(1 - gt  3) or beta-(1 - gt  6) linkages. Additionally, it was observed that the occurrence of different linkages varies during the reaction course, indicating that tailoring favorable GOS structures with improved prebiotic activity can be achieved by adequate control of enzymatic synthesis.",
publisher = "Amer Chemical Soc, Washington",
journal = "Journal of Agricultural and Food Chemistry",
title = "Structural Elucidation of Enzymatically Synthesized Galacto-oligosaccharides Using Ion-Mobility Spectrometry Tandem Mass Spectrometry",
pages = "3615-3609",
number = "18",
volume = "64",
doi = "10.1021/acs.jafc.6b01293"
}

Simović, M., Bezbradica, D., Banjanac, K., Milivojević, A., Fanuel, M., Rogniaux, H., Ropartz, D.,& Veličković, D.. (2016). Structural Elucidation of Enzymatically Synthesized Galacto-oligosaccharides Using Ion-Mobility Spectrometry Tandem Mass Spectrometry. in Journal of Agricultural and Food Chemistry
Amer Chemical Soc, Washington., 64(18), 3609-3615.
https://doi.org/10.1021/acs.jafc.6b01293

Simović M, Bezbradica D, Banjanac K, Milivojević A, Fanuel M, Rogniaux H, Ropartz D, Veličković D. Structural Elucidation of Enzymatically Synthesized Galacto-oligosaccharides Using Ion-Mobility Spectrometry Tandem Mass Spectrometry. in Journal of Agricultural and Food Chemistry. 2016;64(18):3609-3615.
doi:10.1021/acs.jafc.6b01293 .

Simović, Milica, Bezbradica, Dejan, Banjanac, Katarina, Milivojević, Ana, Fanuel, Mathieu, Rogniaux, Helene, Ropartz, David, Veličković, Dušan, "Structural Elucidation of Enzymatically Synthesized Galacto-oligosaccharides Using Ion-Mobility Spectrometry Tandem Mass Spectrometry" in Journal of Agricultural and Food Chemistry, 64, no. 18 (2016):3609-3615,
https://doi.org/10.1021/acs.jafc.6b01293 . .

TY  - JOUR
AU  - Trbojević-Ivić, Jovana
AU  - Dimitrijević, Aleksandra
AU  - Milosavić, Nenad
AU  - Bezbradica, Dejan
AU  - Drakulić, Branko
AU  - Gavrović-Jankulović, Marija
AU  - Pavlović, Marija
AU  - Rogniaux, Helene
AU  - Veličković, Dušan
PY  - 2016
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/3442
AB  - Hydroxyapatite (HAP), a calcium-phosphate bioactive ceramic, is actively employed in medical and separation sciences. Although different classes of biomacromolecules interact with this material, interactions with proteins are the most important, since they directly affect the biocompatibility of the carrier and it's industrial application. In the presented work, we thoroughly investigate and elucidate the interaction mechanism between Candida rugosa lipase (CRL) upon it's immobilization on HAP, since this immobilized enzyme showed advanced catalytic properties in previous studies. Applying elution and protein modification strategies we concluded that Ca-chelation of HAP's C-site and CRL's -COOH groups is the most probable interacting mechanism. A proteomics approach revealed that this chelation is conserved throughout all CRL isoforms, while results of molecular modelling led us to propose the involvement of a specific region of the protein surface and side chains in interactions with HAP.
PB  - Royal Soc Chemistry, Cambridge
T2  - RSC Advances
T1  - Assessment of the interacting mechanism between Candida rugosa lipases and hydroxyapatite and identification of the hydroxyapatite-binding sequence through proteomics and molecular modelling
EP  - 34824
IS  - 41
SP  - 34818
VL  - 6
DO  - 10.1039/c6ra07521e
ER  - 

@article{
author = "Trbojević-Ivić, Jovana and Dimitrijević, Aleksandra and Milosavić, Nenad and Bezbradica, Dejan and Drakulić, Branko and Gavrović-Jankulović, Marija and Pavlović, Marija and Rogniaux, Helene and Veličković, Dušan",
year = "2016",
abstract = "Hydroxyapatite (HAP), a calcium-phosphate bioactive ceramic, is actively employed in medical and separation sciences. Although different classes of biomacromolecules interact with this material, interactions with proteins are the most important, since they directly affect the biocompatibility of the carrier and it's industrial application. In the presented work, we thoroughly investigate and elucidate the interaction mechanism between Candida rugosa lipase (CRL) upon it's immobilization on HAP, since this immobilized enzyme showed advanced catalytic properties in previous studies. Applying elution and protein modification strategies we concluded that Ca-chelation of HAP's C-site and CRL's -COOH groups is the most probable interacting mechanism. A proteomics approach revealed that this chelation is conserved throughout all CRL isoforms, while results of molecular modelling led us to propose the involvement of a specific region of the protein surface and side chains in interactions with HAP.",
publisher = "Royal Soc Chemistry, Cambridge",
journal = "RSC Advances",
title = "Assessment of the interacting mechanism between Candida rugosa lipases and hydroxyapatite and identification of the hydroxyapatite-binding sequence through proteomics and molecular modelling",
pages = "34824-34818",
number = "41",
volume = "6",
doi = "10.1039/c6ra07521e"
}

Trbojević-Ivić, J., Dimitrijević, A., Milosavić, N., Bezbradica, D., Drakulić, B., Gavrović-Jankulović, M., Pavlović, M., Rogniaux, H.,& Veličković, D.. (2016). Assessment of the interacting mechanism between Candida rugosa lipases and hydroxyapatite and identification of the hydroxyapatite-binding sequence through proteomics and molecular modelling. in RSC Advances
Royal Soc Chemistry, Cambridge., 6(41), 34818-34824.
https://doi.org/10.1039/c6ra07521e

Trbojević-Ivić J, Dimitrijević A, Milosavić N, Bezbradica D, Drakulić B, Gavrović-Jankulović M, Pavlović M, Rogniaux H, Veličković D. Assessment of the interacting mechanism between Candida rugosa lipases and hydroxyapatite and identification of the hydroxyapatite-binding sequence through proteomics and molecular modelling. in RSC Advances. 2016;6(41):34818-34824.
doi:10.1039/c6ra07521e .

Trbojević-Ivić, Jovana, Dimitrijević, Aleksandra, Milosavić, Nenad, Bezbradica, Dejan, Drakulić, Branko, Gavrović-Jankulović, Marija, Pavlović, Marija, Rogniaux, Helene, Veličković, Dušan, "Assessment of the interacting mechanism between Candida rugosa lipases and hydroxyapatite and identification of the hydroxyapatite-binding sequence through proteomics and molecular modelling" in RSC Advances, 6, no. 41 (2016):34818-34824,
https://doi.org/10.1039/c6ra07521e . .

TY  - JOUR
AU  - Simović, Milica
AU  - Veličković, Dušan
AU  - Stojanović, Marija
AU  - Milosavić, Nenad
AU  - Rogniaux, Helene
AU  - Ropartz, David
AU  - Bezbradica, Dejan
PY  - 2015
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/3018
AB  - In this study, enzymatic synthesis of galactoside of salicin, compound with potential physiological activity due to structural resemblance with galectin inhibitors, and analgesic and antipyretic properties of salicin, was performed using beta-galactosidase from Aspergillus oryzae. It was determined, using HPLC and ion mobility mass spectrometry, that enzymatic synthesis was highly selective since only one isomer was formed via primary hydroxyl group on glucose moiety of salicin. The optimization of key experimental factors using response surface methodology enabled galactosyl salicin concentration up to 30.8 mM obtained at lactose concentration 40 mM, salicin concentration 110 mM, enzyme amount 360 IU and reaction time 12 h.
PB  - Elsevier Sci Ltd, Oxford
T2  - Process Biochemistry
T1  - Insight in the regioselective enzymatic transgalactosylation of salicin catalyzed by beta-galactosidase from Aspergillus oryzae
EP  - 788
IS  - 5
SP  - 782
VL  - 50
DO  - 10.1016/j.procbio.2015.01.028
ER  - 

@article{
author = "Simović, Milica and Veličković, Dušan and Stojanović, Marija and Milosavić, Nenad and Rogniaux, Helene and Ropartz, David and Bezbradica, Dejan",
year = "2015",
abstract = "In this study, enzymatic synthesis of galactoside of salicin, compound with potential physiological activity due to structural resemblance with galectin inhibitors, and analgesic and antipyretic properties of salicin, was performed using beta-galactosidase from Aspergillus oryzae. It was determined, using HPLC and ion mobility mass spectrometry, that enzymatic synthesis was highly selective since only one isomer was formed via primary hydroxyl group on glucose moiety of salicin. The optimization of key experimental factors using response surface methodology enabled galactosyl salicin concentration up to 30.8 mM obtained at lactose concentration 40 mM, salicin concentration 110 mM, enzyme amount 360 IU and reaction time 12 h.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Process Biochemistry",
title = "Insight in the regioselective enzymatic transgalactosylation of salicin catalyzed by beta-galactosidase from Aspergillus oryzae",
pages = "788-782",
number = "5",
volume = "50",
doi = "10.1016/j.procbio.2015.01.028"
}

Simović, M., Veličković, D., Stojanović, M., Milosavić, N., Rogniaux, H., Ropartz, D.,& Bezbradica, D.. (2015). Insight in the regioselective enzymatic transgalactosylation of salicin catalyzed by beta-galactosidase from Aspergillus oryzae. in Process Biochemistry
Elsevier Sci Ltd, Oxford., 50(5), 782-788.
https://doi.org/10.1016/j.procbio.2015.01.028

Simović M, Veličković D, Stojanović M, Milosavić N, Rogniaux H, Ropartz D, Bezbradica D. Insight in the regioselective enzymatic transgalactosylation of salicin catalyzed by beta-galactosidase from Aspergillus oryzae. in Process Biochemistry. 2015;50(5):782-788.
doi:10.1016/j.procbio.2015.01.028 .

Simović, Milica, Veličković, Dušan, Stojanović, Marija, Milosavić, Nenad, Rogniaux, Helene, Ropartz, David, Bezbradica, Dejan, "Insight in the regioselective enzymatic transgalactosylation of salicin catalyzed by beta-galactosidase from Aspergillus oryzae" in Process Biochemistry, 50, no. 5 (2015):782-788,
https://doi.org/10.1016/j.procbio.2015.01.028 . .