TY  - JOUR
AU  - Ferjančić, Zorana
AU  - Bihelović, Filip
AU  - Vulović, Bojan
AU  - Matović, Radomir
AU  - Trmčić, Milena
AU  - Janković, Aleksandar
AU  - Pavlović, Miloš
AU  - Đurković, Filip T.
AU  - Prodanović, Radivoje
AU  - Đurđević Đelmaš, Aleksandra
AU  - Kaličanin, Nevena
AU  - Zlatović, Mario
AU  - Sladić, Dušan
AU  - Vallet, Thomas
AU  - Vignuzzi, Marco
AU  - Saičić, Radomir N.
PY  - 2024
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7585
AB  - We developed new iminosugar-based glycosidase inhibitors against SARS-CoV-2. Known drugs (miglustat, migalastat, miglitol, and swainsonine) were chosen as lead compounds to develop three classes of glycosidase inhibitors (α-glucosidase, α-galactosidase, and mannosidase). Molecular modelling of the lead compounds, synthesis of the compounds with the highest docking scores, enzyme inhibition tests, and in vitro antiviral assays afforded rationally designed inhibitors. Two highly active α-glucosidase inhibitors were discovered, where one of them is the most potent iminosugar-based anti-SARS-CoV-2 agent to date (EC90 = 1.94 µM in A549-ACE2 cells against Omicron BA.1 strain). However, galactosidase inhibitors did not exhibit antiviral activity, whereas mannosidase inhibitors were both active and cytotoxic. As our iminosugar-based drug candidates act by a host-directed mechanism, they should be more resilient to drug resistance. Moreover, this strategy could be extended to identify potential drug candidates for other viral infections.
PB  - Taylor and Francis Group
T2  - Journal of Enzyme Inhibition and Medicinal Chemistry
T1  - Development of iminosugar-based glycosidase inhibitors as drug candidates for SARS-CoV-2 virus via molecular modelling and in vitro studies
VL  - 39
IS  - 1
SP  - 2289007
DO  - 10.1080/14756366.2023.2289007
ER  - 

@article{
author = "Ferjančić, Zorana and Bihelović, Filip and Vulović, Bojan and Matović, Radomir and Trmčić, Milena and Janković, Aleksandar and Pavlović, Miloš and Đurković, Filip T. and Prodanović, Radivoje and Đurđević Đelmaš, Aleksandra and Kaličanin, Nevena and Zlatović, Mario and Sladić, Dušan and Vallet, Thomas and Vignuzzi, Marco and Saičić, Radomir N.",
year = "2024",
abstract = "We developed new iminosugar-based glycosidase inhibitors against SARS-CoV-2. Known drugs (miglustat, migalastat, miglitol, and swainsonine) were chosen as lead compounds to develop three classes of glycosidase inhibitors (α-glucosidase, α-galactosidase, and mannosidase). Molecular modelling of the lead compounds, synthesis of the compounds with the highest docking scores, enzyme inhibition tests, and in vitro antiviral assays afforded rationally designed inhibitors. Two highly active α-glucosidase inhibitors were discovered, where one of them is the most potent iminosugar-based anti-SARS-CoV-2 agent to date (EC90 = 1.94 µM in A549-ACE2 cells against Omicron BA.1 strain). However, galactosidase inhibitors did not exhibit antiviral activity, whereas mannosidase inhibitors were both active and cytotoxic. As our iminosugar-based drug candidates act by a host-directed mechanism, they should be more resilient to drug resistance. Moreover, this strategy could be extended to identify potential drug candidates for other viral infections.",
publisher = "Taylor and Francis Group",
journal = "Journal of Enzyme Inhibition and Medicinal Chemistry",
title = "Development of iminosugar-based glycosidase inhibitors as drug candidates for SARS-CoV-2 virus via molecular modelling and in vitro studies",
volume = "39",
number = "1",
pages = "2289007",
doi = "10.1080/14756366.2023.2289007"
}

Ferjančić, Z., Bihelović, F., Vulović, B., Matović, R., Trmčić, M., Janković, A., Pavlović, M., Đurković, F. T., Prodanović, R., Đurđević Đelmaš, A., Kaličanin, N., Zlatović, M., Sladić, D., Vallet, T., Vignuzzi, M.,& Saičić, R. N.. (2024). Development of iminosugar-based glycosidase inhibitors as drug candidates for SARS-CoV-2 virus via molecular modelling and in vitro studies. in Journal of Enzyme Inhibition and Medicinal Chemistry
Taylor and Francis Group., 39(1), 2289007.
https://doi.org/10.1080/14756366.2023.2289007

Ferjančić Z, Bihelović F, Vulović B, Matović R, Trmčić M, Janković A, Pavlović M, Đurković FT, Prodanović R, Đurđević Đelmaš A, Kaličanin N, Zlatović M, Sladić D, Vallet T, Vignuzzi M, Saičić RN. Development of iminosugar-based glycosidase inhibitors as drug candidates for SARS-CoV-2 virus via molecular modelling and in vitro studies. in Journal of Enzyme Inhibition and Medicinal Chemistry. 2024;39(1):2289007.
doi:10.1080/14756366.2023.2289007 .

Ferjančić, Zorana, Bihelović, Filip, Vulović, Bojan, Matović, Radomir, Trmčić, Milena, Janković, Aleksandar, Pavlović, Miloš, Đurković, Filip T., Prodanović, Radivoje, Đurđević Đelmaš, Aleksandra, Kaličanin, Nevena, Zlatović, Mario, Sladić, Dušan, Vallet, Thomas, Vignuzzi, Marco, Saičić, Radomir N., "Development of iminosugar-based glycosidase inhibitors as drug candidates for SARS-CoV-2 virus via molecular modelling and in vitro studies" in Journal of Enzyme Inhibition and Medicinal Chemistry, 39, no. 1 (2024):2289007,
https://doi.org/10.1080/14756366.2023.2289007 . .

TY  - JOUR
AU  - Đukić, Ivana
AU  - Kaličanin, Nevena
AU  - Senćanski, Milan
AU  - Pajović, Snežana B.
AU  - Milićević, Jelena
AU  - Prljić, Jelena
AU  - Paessler, Slobodan
AU  - Prodanović, Radivoje
AU  - Glišić, Sanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/5629
AB  - Drug resistance is a critical problem in health care that affects therapy outcomes and requires new approaches to drug
design. SARS-CoV-2 Mpro mutations are of concern as they can potentially reduce therapeutic efficacy. Viral infections are amongst the
many disorders for which nutraceuticals have been employed as an adjunct therapy. The aim of this study was to examine the potential
in vitro activity of L-arginine and vitamin C against SARS-CoV-2 Mpro. Methods: The Mpro inhibition assay was developed by cloning,
expression, purification, and characterization of Mpro. Selected compounds were then screened for protease inhibition. Results: Larginine
was found to be active against SARS-CoV-2 Mpro, while a vitamin C/L-arginine combination had a synergistic antiviral action
against Mpro. These findings confirm the results of our previous in silico repurposing study that showed L-arginine and vitamin C were
potential Mpro inhibitors. Moreover, they suggest a possible molecular mechanism to explain the beneficial effect of arginine in COVID
patients. Conclusions: The findings of the current study are important because they help to identify COVID-19 treatments that are
efficient, inexpensive, and have a favorable safety profile. The results of this study also suggest a possible adjuvant nutritional strategy
for COVID-19 that could be used in conjunction with pharmacological agents.
PB  - IMR Press
T2  - Frontiers in Bioscience Landmark
T1  - Inhibition of SARS-CoV-2 Mpro with Vitamin C, L-Arginine and a Vitamin C/L-Arginine Combination
VL  - 28
IS  - 1
SP  - 8
DO  - 10.31083/j.fbl2801008
ER  - 

@article{
author = "Đukić, Ivana and Kaličanin, Nevena and Senćanski, Milan and Pajović, Snežana B. and Milićević, Jelena and Prljić, Jelena and Paessler, Slobodan and Prodanović, Radivoje and Glišić, Sanja",
year = "2023",
abstract = "Drug resistance is a critical problem in health care that affects therapy outcomes and requires new approaches to drug
design. SARS-CoV-2 Mpro mutations are of concern as they can potentially reduce therapeutic efficacy. Viral infections are amongst the
many disorders for which nutraceuticals have been employed as an adjunct therapy. The aim of this study was to examine the potential
in vitro activity of L-arginine and vitamin C against SARS-CoV-2 Mpro. Methods: The Mpro inhibition assay was developed by cloning,
expression, purification, and characterization of Mpro. Selected compounds were then screened for protease inhibition. Results: Larginine
was found to be active against SARS-CoV-2 Mpro, while a vitamin C/L-arginine combination had a synergistic antiviral action
against Mpro. These findings confirm the results of our previous in silico repurposing study that showed L-arginine and vitamin C were
potential Mpro inhibitors. Moreover, they suggest a possible molecular mechanism to explain the beneficial effect of arginine in COVID
patients. Conclusions: The findings of the current study are important because they help to identify COVID-19 treatments that are
efficient, inexpensive, and have a favorable safety profile. The results of this study also suggest a possible adjuvant nutritional strategy
for COVID-19 that could be used in conjunction with pharmacological agents.",
publisher = "IMR Press",
journal = "Frontiers in Bioscience Landmark",
title = "Inhibition of SARS-CoV-2 Mpro with Vitamin C, L-Arginine and a Vitamin C/L-Arginine Combination",
volume = "28",
number = "1",
pages = "8",
doi = "10.31083/j.fbl2801008"
}

Đukić, I., Kaličanin, N., Senćanski, M., Pajović, S. B., Milićević, J., Prljić, J., Paessler, S., Prodanović, R.,& Glišić, S.. (2023). Inhibition of SARS-CoV-2 Mpro with Vitamin C, L-Arginine and a Vitamin C/L-Arginine Combination. in Frontiers in Bioscience Landmark
IMR Press., 28(1), 8.
https://doi.org/10.31083/j.fbl2801008

Đukić I, Kaličanin N, Senćanski M, Pajović SB, Milićević J, Prljić J, Paessler S, Prodanović R, Glišić S. Inhibition of SARS-CoV-2 Mpro with Vitamin C, L-Arginine and a Vitamin C/L-Arginine Combination. in Frontiers in Bioscience Landmark. 2023;28(1):8.
doi:10.31083/j.fbl2801008 .

Đukić, Ivana, Kaličanin, Nevena, Senćanski, Milan, Pajović, Snežana B., Milićević, Jelena, Prljić, Jelena, Paessler, Slobodan, Prodanović, Radivoje, Glišić, Sanja, "Inhibition of SARS-CoV-2 Mpro with Vitamin C, L-Arginine and a Vitamin C/L-Arginine Combination" in Frontiers in Bioscience Landmark, 28, no. 1 (2023):8,
https://doi.org/10.31083/j.fbl2801008 . .

TY  - JOUR
AU  - Protić, Sara
AU  - Kaličanin, Nevena
AU  - Senćanski, Milan
AU  - Prodanović, Olivera
AU  - Milićević, Jelena
AU  - Perović, Vladimir
AU  - Paessler, Slobodan
AU  - Prodanović, Radivoje
AU  - Glišić, Sanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/5652
AB  - Finding an effective drug to prevent or treat COVID-19 is of utmost importance in tcurrent
pandemic. Since developing a new treatment takes a significant amount of time, drug repurposing
can be an effective option for achieving a rapid response. This study used a combined in silico virtual
screening protocol for candidate SARS-CoV-2 PLpro inhibitors. The Drugbank database was searched
first, using the Informational Spectrum Method for Small Molecules, followed by molecular docking.
Gramicidin D was selected as a peptide drug, showing the best in silico interaction profile with PLpro.
After the expression and purification of PLpro, gramicidin D was screened for protease inhibition
in vitro and was found to be active against PLpro. The current study’s findings are significant
because it is critical to identify COVID-19 therapies that are efficient, affordable, and have a favorable
safety profile.
PB  - Switzerland : Multidisciplinary Digital Publishing Institute (MDPI)
T2  - International Journal of Molecular Sciences
T1  - In Silico and In Vitro Inhibition of SARS-CoV-2 PLpro with Gramicidin D
VL  - 24
IS  - 3
SP  - 1955
DO  - 10.3390/ijms24031955
ER  - 

@article{
author = "Protić, Sara and Kaličanin, Nevena and Senćanski, Milan and Prodanović, Olivera and Milićević, Jelena and Perović, Vladimir and Paessler, Slobodan and Prodanović, Radivoje and Glišić, Sanja",
year = "2023",
abstract = "Finding an effective drug to prevent or treat COVID-19 is of utmost importance in tcurrent
pandemic. Since developing a new treatment takes a significant amount of time, drug repurposing
can be an effective option for achieving a rapid response. This study used a combined in silico virtual
screening protocol for candidate SARS-CoV-2 PLpro inhibitors. The Drugbank database was searched
first, using the Informational Spectrum Method for Small Molecules, followed by molecular docking.
Gramicidin D was selected as a peptide drug, showing the best in silico interaction profile with PLpro.
After the expression and purification of PLpro, gramicidin D was screened for protease inhibition
in vitro and was found to be active against PLpro. The current study’s findings are significant
because it is critical to identify COVID-19 therapies that are efficient, affordable, and have a favorable
safety profile.",
publisher = "Switzerland : Multidisciplinary Digital Publishing Institute (MDPI)",
journal = "International Journal of Molecular Sciences",
title = "In Silico and In Vitro Inhibition of SARS-CoV-2 PLpro with Gramicidin D",
volume = "24",
number = "3",
pages = "1955",
doi = "10.3390/ijms24031955"
}

Protić, S., Kaličanin, N., Senćanski, M., Prodanović, O., Milićević, J., Perović, V., Paessler, S., Prodanović, R.,& Glišić, S.. (2023). In Silico and In Vitro Inhibition of SARS-CoV-2 PLpro with Gramicidin D. in International Journal of Molecular Sciences
Switzerland : Multidisciplinary Digital Publishing Institute (MDPI)., 24(3), 1955.
https://doi.org/10.3390/ijms24031955

Protić S, Kaličanin N, Senćanski M, Prodanović O, Milićević J, Perović V, Paessler S, Prodanović R, Glišić S. In Silico and In Vitro Inhibition of SARS-CoV-2 PLpro with Gramicidin D. in International Journal of Molecular Sciences. 2023;24(3):1955.
doi:10.3390/ijms24031955 .

Protić, Sara, Kaličanin, Nevena, Senćanski, Milan, Prodanović, Olivera, Milićević, Jelena, Perović, Vladimir, Paessler, Slobodan, Prodanović, Radivoje, Glišić, Sanja, "In Silico and In Vitro Inhibition of SARS-CoV-2 PLpro with Gramicidin D" in International Journal of Molecular Sciences, 24, no. 3 (2023):1955,
https://doi.org/10.3390/ijms24031955 . .

TY  - JOUR
AU  - Kaličanin, Nevena
AU  - Balaž, Ana Marija
AU  - Prodanović, Olivera
AU  - Prodanović, Radivoje
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7171
AB  - The aim of this research was to prove the function of the putative opine dehydrogenase from Desulfohalobium retbaense and to characterize the enzyme in terms of functional and kinetic parameters. A putative opine dehydrogenase was identified from a metagenomic library by a sequence-based technique search of the metagenomic library, and afterward was successfully heterologously produced in Escherichia coli. In order to examine its potential for applications in the synthesis of secondary amines, first the substrate specificity of the enzyme towards different amino donors and amino acceptors was determined. The highest affinity was observed towards small amino acids, preferentially L-alanine, and when it comes to α-keto acids, pyruvate proved to be a preferential amino acceptor. The highest activity was observed at pH 6.5 in the absence of salts. The enzyme showed remarkable stability in a wide range of experimental conditions, such as broad pH stability (from 6.0–11.0 after 30 min incubation in buffers at a certain pH), stability in the presence of NaCl up to 3.0 M for 24 h, it retained 80% of the initial activity after 1 h incubation at 45°C, and 65% of the initial activity after 24 h incubation in 30% dimethyl sulfoxide.
PB  - Wiley-VCH GmbH
T2  - ChemBioChem
T1  - Heterologous Expression and Partial Characterization of a Putative Opine Dehydrogenase from a Metagenomic Sequence of Desulfohalobium retbaense
VL  - 24
IS  - 20
IS  - e202300414
DO  - 10.1002/cbic.202300414
ER  - 

@article{
author = "Kaličanin, Nevena and Balaž, Ana Marija and Prodanović, Olivera and Prodanović, Radivoje",
year = "2023",
abstract = "The aim of this research was to prove the function of the putative opine dehydrogenase from Desulfohalobium retbaense and to characterize the enzyme in terms of functional and kinetic parameters. A putative opine dehydrogenase was identified from a metagenomic library by a sequence-based technique search of the metagenomic library, and afterward was successfully heterologously produced in Escherichia coli. In order to examine its potential for applications in the synthesis of secondary amines, first the substrate specificity of the enzyme towards different amino donors and amino acceptors was determined. The highest affinity was observed towards small amino acids, preferentially L-alanine, and when it comes to α-keto acids, pyruvate proved to be a preferential amino acceptor. The highest activity was observed at pH 6.5 in the absence of salts. The enzyme showed remarkable stability in a wide range of experimental conditions, such as broad pH stability (from 6.0–11.0 after 30 min incubation in buffers at a certain pH), stability in the presence of NaCl up to 3.0 M for 24 h, it retained 80% of the initial activity after 1 h incubation at 45°C, and 65% of the initial activity after 24 h incubation in 30% dimethyl sulfoxide.",
publisher = "Wiley-VCH GmbH",
journal = "ChemBioChem",
title = "Heterologous Expression and Partial Characterization of a Putative Opine Dehydrogenase from a Metagenomic Sequence of Desulfohalobium retbaense",
volume = "24",
number = "20, e202300414",
doi = "10.1002/cbic.202300414"
}

Kaličanin, N., Balaž, A. M., Prodanović, O.,& Prodanović, R.. (2023). Heterologous Expression and Partial Characterization of a Putative Opine Dehydrogenase from a Metagenomic Sequence of Desulfohalobium retbaense. in ChemBioChem
Wiley-VCH GmbH., 24(20).
https://doi.org/10.1002/cbic.202300414

Kaličanin N, Balaž AM, Prodanović O, Prodanović R. Heterologous Expression and Partial Characterization of a Putative Opine Dehydrogenase from a Metagenomic Sequence of Desulfohalobium retbaense. in ChemBioChem. 2023;24(20).
doi:10.1002/cbic.202300414 .

Kaličanin, Nevena, Balaž, Ana Marija, Prodanović, Olivera, Prodanović, Radivoje, "Heterologous Expression and Partial Characterization of a Putative Opine Dehydrogenase from a Metagenomic Sequence of Desulfohalobium retbaense" in ChemBioChem, 24, no. 20 (2023),
https://doi.org/10.1002/cbic.202300414 . .

TY  - JOUR
AU  - Kaličanin, Nevena
AU  - Kovačević, Gordana
AU  - Spasojević, Milica
AU  - Prodanović, Olivera
AU  - Jovanović-Šanta, Suzana
AU  - Škorić, Dušan
AU  - Opsenica, Dejan
AU  - Prodanović, Radivoje
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/5370
AB  - The aim of this research was to improve the operational stability and enable the reusability of ω-transaminase for synthesis of new enantiopure chiral amines of steroids. Dihydrotestosterone was used to optimize the synthetic procedure of corresponding amino-steroid on a larger scale. The obtained product 3α-amino-5α-androstan-17β-ol was isolated and characterized. The enzyme was immobilized on a methacrylate-based carrier, giving the specific activity of 1.84 U/g of dry polymer. Higher residual activity of the immobilized enzyme in comparison to the soluble form (100 % versus 35%) after 24 h incubation in 35 % dimethylformamide (DMF) was obtained. The soluble enzyme retained 19 % of the initial activity after 2 h incubation in 35 % DMF at 70 °C, while the activity of the immobilized enzyme decreased only to 75 %. Immobilized retained 85 % of initial activity after ten consecutive cycles of 3α-amino-5α-androstan-17β-ol synthesis. We have tested the specificity of the ArRMut11 variant, further increased its stability by immobilization, and used it in several cycles for the synthesis of 3α-amino-5α-androstan-17β-ol. We showed that the enzyme previously evolved for higher stability as the immobilized variant showed more increased stability and high reusability that can more effectively be applied for the biosynthesis of amino steroids.
PB  - Elsevier
T2  - Process Biochemistry
T1  - Immobilization of ArRMut11 omega-transaminase for increased operational stability and reusability in the synthesis of 3α-amino-5α-androstan-17β-ol
VL  - 121
SP  - 674
EP  - 680
DO  - 10.1016/j.procbio.2022.08.016
ER  - 

@article{
author = "Kaličanin, Nevena and Kovačević, Gordana and Spasojević, Milica and Prodanović, Olivera and Jovanović-Šanta, Suzana and Škorić, Dušan and Opsenica, Dejan and Prodanović, Radivoje",
year = "2022",
abstract = "The aim of this research was to improve the operational stability and enable the reusability of ω-transaminase for synthesis of new enantiopure chiral amines of steroids. Dihydrotestosterone was used to optimize the synthetic procedure of corresponding amino-steroid on a larger scale. The obtained product 3α-amino-5α-androstan-17β-ol was isolated and characterized. The enzyme was immobilized on a methacrylate-based carrier, giving the specific activity of 1.84 U/g of dry polymer. Higher residual activity of the immobilized enzyme in comparison to the soluble form (100 % versus 35%) after 24 h incubation in 35 % dimethylformamide (DMF) was obtained. The soluble enzyme retained 19 % of the initial activity after 2 h incubation in 35 % DMF at 70 °C, while the activity of the immobilized enzyme decreased only to 75 %. Immobilized retained 85 % of initial activity after ten consecutive cycles of 3α-amino-5α-androstan-17β-ol synthesis. We have tested the specificity of the ArRMut11 variant, further increased its stability by immobilization, and used it in several cycles for the synthesis of 3α-amino-5α-androstan-17β-ol. We showed that the enzyme previously evolved for higher stability as the immobilized variant showed more increased stability and high reusability that can more effectively be applied for the biosynthesis of amino steroids.",
publisher = "Elsevier",
journal = "Process Biochemistry",
title = "Immobilization of ArRMut11 omega-transaminase for increased operational stability and reusability in the synthesis of 3α-amino-5α-androstan-17β-ol",
volume = "121",
pages = "674-680",
doi = "10.1016/j.procbio.2022.08.016"
}

Kaličanin, N., Kovačević, G., Spasojević, M., Prodanović, O., Jovanović-Šanta, S., Škorić, D., Opsenica, D.,& Prodanović, R.. (2022). Immobilization of ArRMut11 omega-transaminase for increased operational stability and reusability in the synthesis of 3α-amino-5α-androstan-17β-ol. in Process Biochemistry
Elsevier., 121, 674-680.
https://doi.org/10.1016/j.procbio.2022.08.016

Kaličanin N, Kovačević G, Spasojević M, Prodanović O, Jovanović-Šanta S, Škorić D, Opsenica D, Prodanović R. Immobilization of ArRMut11 omega-transaminase for increased operational stability and reusability in the synthesis of 3α-amino-5α-androstan-17β-ol. in Process Biochemistry. 2022;121:674-680.
doi:10.1016/j.procbio.2022.08.016 .

Kaličanin, Nevena, Kovačević, Gordana, Spasojević, Milica, Prodanović, Olivera, Jovanović-Šanta, Suzana, Škorić, Dušan, Opsenica, Dejan, Prodanović, Radivoje, "Immobilization of ArRMut11 omega-transaminase for increased operational stability and reusability in the synthesis of 3α-amino-5α-androstan-17β-ol" in Process Biochemistry, 121 (2022):674-680,
https://doi.org/10.1016/j.procbio.2022.08.016 . .

TY  - JOUR
AU  - Kaličanin, Nevena
AU  - Kovačević, Gordana
AU  - Spasojević, Milica
AU  - Prodanović, Olivera
AU  - Jovanović-Šanta, Suzana
AU  - Škorić, Dušan
AU  - Opsenica, Dejan
AU  - Prodanović, Radivoje
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/5545
AB  - The aim of this research was to improve the operational stability and enable the reusability of ω-transaminase for synthesis of new enantiopure chiral amines of steroids. Dihydrotestosterone was used to optimize the synthetic procedure of corresponding amino-steroid on a larger scale. The obtained product 3α-amino-5α-androstan-17β-ol was isolated and characterized. The enzyme was immobilized on a methacrylate-based carrier, giving the specific activity of 1.84 U/g of dry polymer. Higher residual activity of the immobilized enzyme in comparison to the soluble form (100 % versus 35%) after 24 h incubation in 35 % dimethylformamide (DMF) was obtained. The soluble enzyme retained 19 % of the initial activity after 2 h incubation in 35 % DMF at 70 °C, while the activity of the immobilized enzyme decreased only to 75 %. Immobilized retained 85 % of initial activity after ten consecutive cycles of 3α-amino-5α-androstan-17β-ol synthesis. We have tested the specificity of the ArRMut11 variant, further increased its stability by immobilization, and used it in several cycles for the synthesis of 3α-amino-5α-androstan-17β-ol. We showed that the enzyme previously evolved for higher stability as the immobilized variant showed more increased stability and high reusability that can more effectively be applied for the biosynthesis of amino steroids.
PB  - Elsevier
T2  - Process Biochemistry
T1  - Immobilization of ArRMut11 omega-transaminase for increased operational stability and reusability in the synthesis of 3α-amino-5α-androstan-17β-ol
VL  - 121
SP  - 674
EP  - 680
DO  - 10.1016/j.procbio.2022.08.016
ER  - 

@article{
author = "Kaličanin, Nevena and Kovačević, Gordana and Spasojević, Milica and Prodanović, Olivera and Jovanović-Šanta, Suzana and Škorić, Dušan and Opsenica, Dejan and Prodanović, Radivoje",
year = "2022",
abstract = "The aim of this research was to improve the operational stability and enable the reusability of ω-transaminase for synthesis of new enantiopure chiral amines of steroids. Dihydrotestosterone was used to optimize the synthetic procedure of corresponding amino-steroid on a larger scale. The obtained product 3α-amino-5α-androstan-17β-ol was isolated and characterized. The enzyme was immobilized on a methacrylate-based carrier, giving the specific activity of 1.84 U/g of dry polymer. Higher residual activity of the immobilized enzyme in comparison to the soluble form (100 % versus 35%) after 24 h incubation in 35 % dimethylformamide (DMF) was obtained. The soluble enzyme retained 19 % of the initial activity after 2 h incubation in 35 % DMF at 70 °C, while the activity of the immobilized enzyme decreased only to 75 %. Immobilized retained 85 % of initial activity after ten consecutive cycles of 3α-amino-5α-androstan-17β-ol synthesis. We have tested the specificity of the ArRMut11 variant, further increased its stability by immobilization, and used it in several cycles for the synthesis of 3α-amino-5α-androstan-17β-ol. We showed that the enzyme previously evolved for higher stability as the immobilized variant showed more increased stability and high reusability that can more effectively be applied for the biosynthesis of amino steroids.",
publisher = "Elsevier",
journal = "Process Biochemistry",
title = "Immobilization of ArRMut11 omega-transaminase for increased operational stability and reusability in the synthesis of 3α-amino-5α-androstan-17β-ol",
volume = "121",
pages = "674-680",
doi = "10.1016/j.procbio.2022.08.016"
}

Kaličanin, N., Kovačević, G., Spasojević, M., Prodanović, O., Jovanović-Šanta, S., Škorić, D., Opsenica, D.,& Prodanović, R.. (2022). Immobilization of ArRMut11 omega-transaminase for increased operational stability and reusability in the synthesis of 3α-amino-5α-androstan-17β-ol. in Process Biochemistry
Elsevier., 121, 674-680.
https://doi.org/10.1016/j.procbio.2022.08.016

Kaličanin N, Kovačević G, Spasojević M, Prodanović O, Jovanović-Šanta S, Škorić D, Opsenica D, Prodanović R. Immobilization of ArRMut11 omega-transaminase for increased operational stability and reusability in the synthesis of 3α-amino-5α-androstan-17β-ol. in Process Biochemistry. 2022;121:674-680.
doi:10.1016/j.procbio.2022.08.016 .

Kaličanin, Nevena, Kovačević, Gordana, Spasojević, Milica, Prodanović, Olivera, Jovanović-Šanta, Suzana, Škorić, Dušan, Opsenica, Dejan, Prodanović, Radivoje, "Immobilization of ArRMut11 omega-transaminase for increased operational stability and reusability in the synthesis of 3α-amino-5α-androstan-17β-ol" in Process Biochemistry, 121 (2022):674-680,
https://doi.org/10.1016/j.procbio.2022.08.016 . .

TY  - CONF
AU  - Kaličanin, Nevena
AU  - Balaž, Ana Marija
AU  - Prodanović, Olivera
AU  - Prodanović, Radivoje
PY  - 2022
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7099
AB  - Opine dehydrogenases are a family of NAD(P)H dependent oxidoreductases, whichcatalyze the reductive condensation of an α amino group from an amino acid with an αketo acid during anaerobic glycolysis by regenerating NAD. They are widespread incephalopods and mollusks. Opines are associated with crown gall tumor pathogenesiscaused by A. tumefaciens providing nutrients to the pathogen, and novel opine compoundsacting as metallophores have been identified. Besides, opine-type secondary aminedicarboxylic acids are chiral intermediates of angiotensin-converting enzyme inhibitors. Anovel enzyme originating from an extremophile bacterium, with assumed opinedehydrogenase function was successfully expressed in Escherichia coli STAR cells andpurified by affinity chromatography. Molecular mass determined by SDS-PAGE wasapproximately 40 kDa. The activity was measured by using pyruvate and alanine assubstrates, by which proved that it has opine dehydrogenase activity.
PB  - Faculty of Chemistry
PB  - Serbian Biochemical Society
C3  - Proceedings - Eleventh Conference, Scientific meeting of an international character "Amazing Biochemistry", September 22nd and 23rd, 2022, Novi Sad, Serbia
T1  - Production of a novel opine dehydrogenase
SP  - 79
EP  - 79
UR  - https://hdl.handle.net/21.15107/rcub_cer_7099
ER  - 

@conference{
author = "Kaličanin, Nevena and Balaž, Ana Marija and Prodanović, Olivera and Prodanović, Radivoje",
year = "2022",
abstract = "Opine dehydrogenases are a family of NAD(P)H dependent oxidoreductases, whichcatalyze the reductive condensation of an α amino group from an amino acid with an αketo acid during anaerobic glycolysis by regenerating NAD. They are widespread incephalopods and mollusks. Opines are associated with crown gall tumor pathogenesiscaused by A. tumefaciens providing nutrients to the pathogen, and novel opine compoundsacting as metallophores have been identified. Besides, opine-type secondary aminedicarboxylic acids are chiral intermediates of angiotensin-converting enzyme inhibitors. Anovel enzyme originating from an extremophile bacterium, with assumed opinedehydrogenase function was successfully expressed in Escherichia coli STAR cells andpurified by affinity chromatography. Molecular mass determined by SDS-PAGE wasapproximately 40 kDa. The activity was measured by using pyruvate and alanine assubstrates, by which proved that it has opine dehydrogenase activity.",
publisher = "Faculty of Chemistry, Serbian Biochemical Society",
journal = "Proceedings - Eleventh Conference, Scientific meeting of an international character "Amazing Biochemistry", September 22nd and 23rd, 2022, Novi Sad, Serbia",
title = "Production of a novel opine dehydrogenase",
pages = "79-79",
url = "https://hdl.handle.net/21.15107/rcub_cer_7099"
}

Kaličanin, N., Balaž, A. M., Prodanović, O.,& Prodanović, R.. (2022). Production of a novel opine dehydrogenase. in Proceedings - Eleventh Conference, Scientific meeting of an international character "Amazing Biochemistry", September 22nd and 23rd, 2022, Novi Sad, Serbia
Faculty of Chemistry., 79-79.
https://hdl.handle.net/21.15107/rcub_cer_7099

Kaličanin N, Balaž AM, Prodanović O, Prodanović R. Production of a novel opine dehydrogenase. in Proceedings - Eleventh Conference, Scientific meeting of an international character "Amazing Biochemistry", September 22nd and 23rd, 2022, Novi Sad, Serbia. 2022;:79-79.
https://hdl.handle.net/21.15107/rcub_cer_7099 .

Kaličanin, Nevena, Balaž, Ana Marija, Prodanović, Olivera, Prodanović, Radivoje, "Production of a novel opine dehydrogenase" in Proceedings - Eleventh Conference, Scientific meeting of an international character "Amazing Biochemistry", September 22nd and 23rd, 2022, Novi Sad, Serbia (2022):79-79,
https://hdl.handle.net/21.15107/rcub_cer_7099 .

TY  - CONF
AU  - Marković, Nevena
AU  - Jovanović Šanta, Suzana
AU  - Prodanović, Radivoje
PY  - 2017
UR  - http://www.bds.org.rs/en/conferences.php
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/3281
AB  - Transaminases (EC 2.6.1.X) are enzymes which catalyze reversible transfer of amino group from amino acids to α-keto acids by using piridoxal-5ʼ-phosphate as a coenzyme. There is a huge interest for the application of ω-transaminases in industrial production of chiral amines and alkaloids since those compounds are extensively used in pharmaceutical, agricultural, and chemical industries. Application of ω-transaminases in asimetric synthesis of these compounds enables efficient production of biologicaly active amines, due to their catalytic properties for synthesis with a high level of enantioselectivity, supstrate promiscuity (they are capable to aminate keto acids, aldehydes and ketones), high turnover number, no requirement for regeneration of external cofactors, and among other cheaper, simpler and green process of production. We are developing biocatalytic route for the synthesis of amino steroids by using ω- transaminase, (R)-selective, ATA-117 enzyme variant from Arthrobacter sp 3. It can be observed that enzyme expression was done in Echerichia coli BL21 D3 pLysS (Figure 1), and HPLC analysis of enzyme activity and specificity toward 15 structuraly different steroid compounds was performed. (R)-methylbenzylamine was used as amino group donor and pyridoxal-5ʼ-phosphate as cofactor. Activity of the enzyme was measured in bacterial lysate based on the absorbance of acetophenone, that is formed during the transamination reaction of (R)-methylbenzylamine. Figures 2 and 3 are showing chromatograms of acetophenone standard and products of reaction performed with enzyme expressed in E. coli and 16,17-epoxypregnenolone. Reactions were analysed on reversed phase column NucleosilC18. Based on the results, we have selected four steroid compounds for which enzyme showed highest activity and with a potential for biological activity. The next step was optimisation of the reaction conditions with a low cost amino donor isopropylamine, and isolation and characterisation of a pure amino steroid products. Until now we have managed to enzymatically synthesize and purify one amino steroid which should be further analysed by spectral characterization and its biological activity will be determined.
PB  - Faculty of Chemistry, Serbian Biochemical Society
C3  - Serbian Biochemical Society Seventh Conference "Biochemistry of Control in Life and Technology" - Proceedings
T1  - Synthesis of potential pharmaceutical active ingredients using omega-transaminase
UR  - https://hdl.handle.net/21.15107/rcub_cer_3281
ER  - 

@conference{
author = "Marković, Nevena and Jovanović Šanta, Suzana and Prodanović, Radivoje",
year = "2017",
abstract = "Transaminases (EC 2.6.1.X) are enzymes which catalyze reversible transfer of amino group from amino acids to α-keto acids by using piridoxal-5ʼ-phosphate as a coenzyme. There is a huge interest for the application of ω-transaminases in industrial production of chiral amines and alkaloids since those compounds are extensively used in pharmaceutical, agricultural, and chemical industries. Application of ω-transaminases in asimetric synthesis of these compounds enables efficient production of biologicaly active amines, due to their catalytic properties for synthesis with a high level of enantioselectivity, supstrate promiscuity (they are capable to aminate keto acids, aldehydes and ketones), high turnover number, no requirement for regeneration of external cofactors, and among other cheaper, simpler and green process of production. We are developing biocatalytic route for the synthesis of amino steroids by using ω- transaminase, (R)-selective, ATA-117 enzyme variant from Arthrobacter sp 3. It can be observed that enzyme expression was done in Echerichia coli BL21 D3 pLysS (Figure 1), and HPLC analysis of enzyme activity and specificity toward 15 structuraly different steroid compounds was performed. (R)-methylbenzylamine was used as amino group donor and pyridoxal-5ʼ-phosphate as cofactor. Activity of the enzyme was measured in bacterial lysate based on the absorbance of acetophenone, that is formed during the transamination reaction of (R)-methylbenzylamine. Figures 2 and 3 are showing chromatograms of acetophenone standard and products of reaction performed with enzyme expressed in E. coli and 16,17-epoxypregnenolone. Reactions were analysed on reversed phase column NucleosilC18. Based on the results, we have selected four steroid compounds for which enzyme showed highest activity and with a potential for biological activity. The next step was optimisation of the reaction conditions with a low cost amino donor isopropylamine, and isolation and characterisation of a pure amino steroid products. Until now we have managed to enzymatically synthesize and purify one amino steroid which should be further analysed by spectral characterization and its biological activity will be determined.",
publisher = "Faculty of Chemistry, Serbian Biochemical Society",
journal = "Serbian Biochemical Society Seventh Conference "Biochemistry of Control in Life and Technology" - Proceedings",
title = "Synthesis of potential pharmaceutical active ingredients using omega-transaminase",
url = "https://hdl.handle.net/21.15107/rcub_cer_3281"
}

Marković, N., Jovanović Šanta, S.,& Prodanović, R.. (2017). Synthesis of potential pharmaceutical active ingredients using omega-transaminase. in Serbian Biochemical Society Seventh Conference "Biochemistry of Control in Life and Technology" - Proceedings
Faculty of Chemistry, Serbian Biochemical Society..
https://hdl.handle.net/21.15107/rcub_cer_3281

Marković N, Jovanović Šanta S, Prodanović R. Synthesis of potential pharmaceutical active ingredients using omega-transaminase. in Serbian Biochemical Society Seventh Conference "Biochemistry of Control in Life and Technology" - Proceedings. 2017;.
https://hdl.handle.net/21.15107/rcub_cer_3281 .

Marković, Nevena, Jovanović Šanta, Suzana, Prodanović, Radivoje, "Synthesis of potential pharmaceutical active ingredients using omega-transaminase" in Serbian Biochemical Society Seventh Conference "Biochemistry of Control in Life and Technology" - Proceedings (2017),
https://hdl.handle.net/21.15107/rcub_cer_3281 .

TY  - JOUR
AU  - Prodanović, Olivera
AU  - Spasojevic, Dragica
AU  - Prokopijevic, Milos
AU  - Radotić, Ksenija
AU  - Marković, Nevena
AU  - Blažić, Marija
AU  - Prodanović, Radivoje
PY  - 2015
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/1727
AB  - Phenol and amino groups were introduced into alginate to different degrees via oxidation with 2.5, 5, 10, 15 and 20 mol% of periodate and reductive amination by tyramine. Modification of alginate with tyramine was confirmed by FTIR spectroscopy and UV-VIS spectroscopy, while concentration of phenol and ionizable groups was determined using absorbance at 275 nm and acid-base titration. All tyramine-alginates were able to form hydrogels after cross-linking with horse radish peroxidase (HRP) and hydrogen peroxide. Tyramine-alginates oxidized with up to 10 mol% of periodate were also capable of forming hydrogels with calcium ions. Tyramine-alginates were tested for HRP immobilization within micro-beads obtained by peroxidase catalyzed droplet polymerization using internal delivery of hydrogen peroxide via glucose oxidase and glucose. Highest activity of immobilized peroxidase was obtained with 20% (w/v) tyramine-alginate obtained via 20 mol% periodate oxidation. Immobilized enzyme was not leaking from the micro-beads and was further kinetically characterized for pyrogallol oxidation. Km for pyrogallol was increased after immobilization from 1.93 mM for soluble HRP to 734 mM for immobilized HRP. The optimum pH was also increased from pH 7.0 to 8.0. Temperature and organic solvent stability improved significantly after immobilization, so that half-life at 70 degrees C increased around four times, while half-life in 80% (v/v) dioxane increased 22 times. After repeated use of 6 times in batch reactor for pyrogallol oxidation immobilized HRP retained 45% of original activity.
PB  - Elsevier
T2  - Reactive and Functional Polymers
T1  - Tyramine modified alginates via periodate oxidation for peroxidase induced hydrogel formation and immobilization
VL  - 93
SP  - 77
EP  - 83
DO  - 10.1016/j.reactfunctpolym.2015.06.004
ER  - 

@article{
author = "Prodanović, Olivera and Spasojevic, Dragica and Prokopijevic, Milos and Radotić, Ksenija and Marković, Nevena and Blažić, Marija and Prodanović, Radivoje",
year = "2015",
abstract = "Phenol and amino groups were introduced into alginate to different degrees via oxidation with 2.5, 5, 10, 15 and 20 mol% of periodate and reductive amination by tyramine. Modification of alginate with tyramine was confirmed by FTIR spectroscopy and UV-VIS spectroscopy, while concentration of phenol and ionizable groups was determined using absorbance at 275 nm and acid-base titration. All tyramine-alginates were able to form hydrogels after cross-linking with horse radish peroxidase (HRP) and hydrogen peroxide. Tyramine-alginates oxidized with up to 10 mol% of periodate were also capable of forming hydrogels with calcium ions. Tyramine-alginates were tested for HRP immobilization within micro-beads obtained by peroxidase catalyzed droplet polymerization using internal delivery of hydrogen peroxide via glucose oxidase and glucose. Highest activity of immobilized peroxidase was obtained with 20% (w/v) tyramine-alginate obtained via 20 mol% periodate oxidation. Immobilized enzyme was not leaking from the micro-beads and was further kinetically characterized for pyrogallol oxidation. Km for pyrogallol was increased after immobilization from 1.93 mM for soluble HRP to 734 mM for immobilized HRP. The optimum pH was also increased from pH 7.0 to 8.0. Temperature and organic solvent stability improved significantly after immobilization, so that half-life at 70 degrees C increased around four times, while half-life in 80% (v/v) dioxane increased 22 times. After repeated use of 6 times in batch reactor for pyrogallol oxidation immobilized HRP retained 45% of original activity.",
publisher = "Elsevier",
journal = "Reactive and Functional Polymers",
title = "Tyramine modified alginates via periodate oxidation for peroxidase induced hydrogel formation and immobilization",
volume = "93",
pages = "77-83",
doi = "10.1016/j.reactfunctpolym.2015.06.004"
}

Prodanović, O., Spasojevic, D., Prokopijevic, M., Radotić, K., Marković, N., Blažić, M.,& Prodanović, R.. (2015). Tyramine modified alginates via periodate oxidation for peroxidase induced hydrogel formation and immobilization. in Reactive and Functional Polymers
Elsevier., 93, 77-83.
https://doi.org/10.1016/j.reactfunctpolym.2015.06.004

Prodanović O, Spasojevic D, Prokopijevic M, Radotić K, Marković N, Blažić M, Prodanović R. Tyramine modified alginates via periodate oxidation for peroxidase induced hydrogel formation and immobilization. in Reactive and Functional Polymers. 2015;93:77-83.
doi:10.1016/j.reactfunctpolym.2015.06.004 .

Prodanović, Olivera, Spasojevic, Dragica, Prokopijevic, Milos, Radotić, Ksenija, Marković, Nevena, Blažić, Marija, Prodanović, Radivoje, "Tyramine modified alginates via periodate oxidation for peroxidase induced hydrogel formation and immobilization" in Reactive and Functional Polymers, 93 (2015):77-83,
https://doi.org/10.1016/j.reactfunctpolym.2015.06.004 . .