TY  - JOUR
AU  - Lazić, Jelena O.
AU  - Spasić, Jelena
AU  - Francuski, Đorđe
AU  - Tokić-Vujošević, Zorana
AU  - Nikodinović-Runić, Jasmina
AU  - Maslak, Veselin
AU  - Đokić, Lidija
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2324
AB  - Michael addition of aldehydes to nitro-olefins is an effective method to obtain useful chiral gamma-nitroaldehydes. gamma-Nitroaldehydes are precursors for chiral gamma-aminobutyric acid analogues, which have numerous pharmacological activities and are used for the treatment of neurological disorders. A whole-cell system based on recombinantly expressed 4-oxalocrotonate tautomerase (4-OT) was developed and shown to be an effective biocatalyst for the Michael addition of branched aldehydes to beta-nitrostyrenes. The aim of this study was to investigate the influence of the substitution of the N-terminal proline with lysine and arginine, both containing a reactive epsilon-amino group, on the Michael addition catalyzed by 4-OT. First, the effects of these mutations were examined by in silico analysis, followed by the generation of three terminal lysine mutants. The generated mutants, 4-OT_K, 4-OT_PK and 4-OT_KK were tested for their ability to utilise beta-nitrostyrene (1), (E)-1-nitro-2-(2-thienyl)ethene (2) and trans-p-chloro-beta-nitrostyrene (3) as Michael acceptors with isobutanal (2-methylpropanal) as the donor. For comparison, the lithium salt of lysine was used in the same organocatalytic reactions. In general, the introduction of lysine had a negative effect on Michael additions based on overall product yields. However, additional lysine residues at the N-terminus of the protein resulted in structural changes that enhanced the activity towards 2 and 3. Therefore, the N-terminal proline is important for 4-OT-catalysed Michael-additions, but it is not essential.
PB  - Serbian Chemical Soc, Belgrade
T2  - Journal of the Serbian Chemical Society
T1  - Importance of N-terminal proline for the promiscuous activity of 4-oxalocrotonate tautomerase (4-OT)
VL  - 81
IS  - 8
SP  - 871
EP  - 881
DO  - 10.2298/JSC160222053L
ER  - 

@article{
author = "Lazić, Jelena O. and Spasić, Jelena and Francuski, Đorđe and Tokić-Vujošević, Zorana and Nikodinović-Runić, Jasmina and Maslak, Veselin and Đokić, Lidija",
year = "2016",
abstract = "Michael addition of aldehydes to nitro-olefins is an effective method to obtain useful chiral gamma-nitroaldehydes. gamma-Nitroaldehydes are precursors for chiral gamma-aminobutyric acid analogues, which have numerous pharmacological activities and are used for the treatment of neurological disorders. A whole-cell system based on recombinantly expressed 4-oxalocrotonate tautomerase (4-OT) was developed and shown to be an effective biocatalyst for the Michael addition of branched aldehydes to beta-nitrostyrenes. The aim of this study was to investigate the influence of the substitution of the N-terminal proline with lysine and arginine, both containing a reactive epsilon-amino group, on the Michael addition catalyzed by 4-OT. First, the effects of these mutations were examined by in silico analysis, followed by the generation of three terminal lysine mutants. The generated mutants, 4-OT_K, 4-OT_PK and 4-OT_KK were tested for their ability to utilise beta-nitrostyrene (1), (E)-1-nitro-2-(2-thienyl)ethene (2) and trans-p-chloro-beta-nitrostyrene (3) as Michael acceptors with isobutanal (2-methylpropanal) as the donor. For comparison, the lithium salt of lysine was used in the same organocatalytic reactions. In general, the introduction of lysine had a negative effect on Michael additions based on overall product yields. However, additional lysine residues at the N-terminus of the protein resulted in structural changes that enhanced the activity towards 2 and 3. Therefore, the N-terminal proline is important for 4-OT-catalysed Michael-additions, but it is not essential.",
publisher = "Serbian Chemical Soc, Belgrade",
journal = "Journal of the Serbian Chemical Society",
title = "Importance of N-terminal proline for the promiscuous activity of 4-oxalocrotonate tautomerase (4-OT)",
volume = "81",
number = "8",
pages = "871-881",
doi = "10.2298/JSC160222053L"
}

Lazić, J. O., Spasić, J., Francuski, Đ., Tokić-Vujošević, Z., Nikodinović-Runić, J., Maslak, V.,& Đokić, L.. (2016). Importance of N-terminal proline for the promiscuous activity of 4-oxalocrotonate tautomerase (4-OT). in Journal of the Serbian Chemical Society
Serbian Chemical Soc, Belgrade., 81(8), 871-881.
https://doi.org/10.2298/JSC160222053L

Lazić JO, Spasić J, Francuski Đ, Tokić-Vujošević Z, Nikodinović-Runić J, Maslak V, Đokić L. Importance of N-terminal proline for the promiscuous activity of 4-oxalocrotonate tautomerase (4-OT). in Journal of the Serbian Chemical Society. 2016;81(8):871-881.
doi:10.2298/JSC160222053L .

Lazić, Jelena O., Spasić, Jelena, Francuski, Đorđe, Tokić-Vujošević, Zorana, Nikodinović-Runić, Jasmina, Maslak, Veselin, Đokić, Lidija, "Importance of N-terminal proline for the promiscuous activity of 4-oxalocrotonate tautomerase (4-OT)" in Journal of the Serbian Chemical Society, 81, no. 8 (2016):871-881,
https://doi.org/10.2298/JSC160222053L . .

TY  - DATA
AU  - Narančić, Tanja
AU  - Kadivojević, Jelena
AU  - Jovanović, Predrag M.
AU  - Francuski, Đorđe
AU  - Bigović, Miljan
AU  - Maslak, Veselin
AU  - Savić, Vladimir
AU  - Vasiljević, Branka
AU  - O'Connor, Kevin E.
AU  - Nikodinović-Runić, Jasmina
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3499
PB  - Elsevier Sci Ltd, Oxford
T2  - Bioresource Technology
T1  - Supplementary data for article: Narančić, T.; Kadivojevic, J.; Jovanovic, P.; Francuski, D.; Bigović, M.; Maslak, V.; Savić, V.; Vasiljević, B.; O’Connor, K. E.; Nikodinović-Runić, J. Highly Efficient Michael-Type Addition of Acetaldehyde to Beta-Nitrostyrenes by Whole Resting Cells of Escherichia Coli Expressing 4-Oxalocrotonate Tautomerase. Bioresource Technology 2013, 142, 462–468. https://doi.org/10.1016/j.biortech.2013.05.074
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3499
ER  - 

@misc{
author = "Narančić, Tanja and Kadivojević, Jelena and Jovanović, Predrag M. and Francuski, Đorđe and Bigović, Miljan and Maslak, Veselin and Savić, Vladimir and Vasiljević, Branka and O'Connor, Kevin E. and Nikodinović-Runić, Jasmina",
year = "2013",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Bioresource Technology",
title = "Supplementary data for article: Narančić, T.; Kadivojevic, J.; Jovanovic, P.; Francuski, D.; Bigović, M.; Maslak, V.; Savić, V.; Vasiljević, B.; O’Connor, K. E.; Nikodinović-Runić, J. Highly Efficient Michael-Type Addition of Acetaldehyde to Beta-Nitrostyrenes by Whole Resting Cells of Escherichia Coli Expressing 4-Oxalocrotonate Tautomerase. Bioresource Technology 2013, 142, 462–468. https://doi.org/10.1016/j.biortech.2013.05.074",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3499"
}

Narančić, T., Kadivojević, J., Jovanović, P. M., Francuski, Đ., Bigović, M., Maslak, V., Savić, V., Vasiljević, B., O'Connor, K. E.,& Nikodinović-Runić, J.. (2013). Supplementary data for article: Narančić, T.; Kadivojevic, J.; Jovanovic, P.; Francuski, D.; Bigović, M.; Maslak, V.; Savić, V.; Vasiljević, B.; O’Connor, K. E.; Nikodinović-Runić, J. Highly Efficient Michael-Type Addition of Acetaldehyde to Beta-Nitrostyrenes by Whole Resting Cells of Escherichia Coli Expressing 4-Oxalocrotonate Tautomerase. Bioresource Technology 2013, 142, 462–468. https://doi.org/10.1016/j.biortech.2013.05.074. in Bioresource Technology
Elsevier Sci Ltd, Oxford..
https://hdl.handle.net/21.15107/rcub_cherry_3499

Narančić T, Kadivojević J, Jovanović PM, Francuski Đ, Bigović M, Maslak V, Savić V, Vasiljević B, O'Connor KE, Nikodinović-Runić J. Supplementary data for article: Narančić, T.; Kadivojevic, J.; Jovanovic, P.; Francuski, D.; Bigović, M.; Maslak, V.; Savić, V.; Vasiljević, B.; O’Connor, K. E.; Nikodinović-Runić, J. Highly Efficient Michael-Type Addition of Acetaldehyde to Beta-Nitrostyrenes by Whole Resting Cells of Escherichia Coli Expressing 4-Oxalocrotonate Tautomerase. Bioresource Technology 2013, 142, 462–468. https://doi.org/10.1016/j.biortech.2013.05.074. in Bioresource Technology. 2013;.
https://hdl.handle.net/21.15107/rcub_cherry_3499 .

Narančić, Tanja, Kadivojević, Jelena, Jovanović, Predrag M., Francuski, Đorđe, Bigović, Miljan, Maslak, Veselin, Savić, Vladimir, Vasiljević, Branka, O'Connor, Kevin E., Nikodinović-Runić, Jasmina, "Supplementary data for article: Narančić, T.; Kadivojevic, J.; Jovanovic, P.; Francuski, D.; Bigović, M.; Maslak, V.; Savić, V.; Vasiljević, B.; O’Connor, K. E.; Nikodinović-Runić, J. Highly Efficient Michael-Type Addition of Acetaldehyde to Beta-Nitrostyrenes by Whole Resting Cells of Escherichia Coli Expressing 4-Oxalocrotonate Tautomerase. Bioresource Technology 2013, 142, 462–468. https://doi.org/10.1016/j.biortech.2013.05.074" in Bioresource Technology (2013),
https://hdl.handle.net/21.15107/rcub_cherry_3499 .

TY  - JOUR
AU  - Narančić, Tanja
AU  - Kadivojević, Jelena
AU  - Jovanović, Predrag M.
AU  - Francuski, Đorđe
AU  - Bigović, Miljan
AU  - Maslak, Veselin
AU  - Savić, Vladimir
AU  - Vasiljević, Branka
AU  - O'Connor, Kevin E.
AU  - Nikodinović-Runić, Jasmina
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1377
AB  - A novel whole cell system based on recombinantly expressed 4-oxalocrotonate tautomerase (4-OT) was developed and shown to be an effective biocatalyst for the asymmetric Michael addition of acetaldehyde to beta-nitrostyrenes. Optimal ratio of substrates (2 mM beta-nitrostyrenes and 20 mM acetaldehyde) and biocatalyst of 5 g of cell dry weight of biocatalyst per liter was determined. Through further bioprocess improvement by sequential addition of substrate 10 mM nitrostyrene biotransformation was achieved within 150 min. Excellent enantioselectivity ( gt 99% ee) and product yields of up to 60% were obtained with beta-nitrostyrene substrate. The biotransformation product, 4-nitro-3-phenyl-butanal, was isolated from aqueous media and further transformed into the corresponding amino alcohol. The biocatalyst exhibited lower reaction rates with p-Cl-, o-Cl- and p-F-beta-nitrostyrenes with product yields of 38%, 51%, 31% and ee values of 84%, 88% and 94% respectively. The importance of the terminal,proline of 4-UT was confirmed by two proline enriched variants and homology modeling.
PB  - Elsevier Sci Ltd, Oxford
T2  - Bioresource Technology
T1  - Highly efficient Michael-type addition of acetaldehyde to beta-nitrostyrenes by whole resting cells of Escherichia coli expressing 4-oxalocrotonate tautomerase
VL  - 142
SP  - 462
EP  - 468
DO  - 10.1016/j.biortech.2013.05.074
ER  - 

@article{
author = "Narančić, Tanja and Kadivojević, Jelena and Jovanović, Predrag M. and Francuski, Đorđe and Bigović, Miljan and Maslak, Veselin and Savić, Vladimir and Vasiljević, Branka and O'Connor, Kevin E. and Nikodinović-Runić, Jasmina",
year = "2013",
abstract = "A novel whole cell system based on recombinantly expressed 4-oxalocrotonate tautomerase (4-OT) was developed and shown to be an effective biocatalyst for the asymmetric Michael addition of acetaldehyde to beta-nitrostyrenes. Optimal ratio of substrates (2 mM beta-nitrostyrenes and 20 mM acetaldehyde) and biocatalyst of 5 g of cell dry weight of biocatalyst per liter was determined. Through further bioprocess improvement by sequential addition of substrate 10 mM nitrostyrene biotransformation was achieved within 150 min. Excellent enantioselectivity ( gt 99% ee) and product yields of up to 60% were obtained with beta-nitrostyrene substrate. The biotransformation product, 4-nitro-3-phenyl-butanal, was isolated from aqueous media and further transformed into the corresponding amino alcohol. The biocatalyst exhibited lower reaction rates with p-Cl-, o-Cl- and p-F-beta-nitrostyrenes with product yields of 38%, 51%, 31% and ee values of 84%, 88% and 94% respectively. The importance of the terminal,proline of 4-UT was confirmed by two proline enriched variants and homology modeling.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Bioresource Technology",
title = "Highly efficient Michael-type addition of acetaldehyde to beta-nitrostyrenes by whole resting cells of Escherichia coli expressing 4-oxalocrotonate tautomerase",
volume = "142",
pages = "462-468",
doi = "10.1016/j.biortech.2013.05.074"
}

Narančić, T., Kadivojević, J., Jovanović, P. M., Francuski, Đ., Bigović, M., Maslak, V., Savić, V., Vasiljević, B., O'Connor, K. E.,& Nikodinović-Runić, J.. (2013). Highly efficient Michael-type addition of acetaldehyde to beta-nitrostyrenes by whole resting cells of Escherichia coli expressing 4-oxalocrotonate tautomerase. in Bioresource Technology
Elsevier Sci Ltd, Oxford., 142, 462-468.
https://doi.org/10.1016/j.biortech.2013.05.074

Narančić T, Kadivojević J, Jovanović PM, Francuski Đ, Bigović M, Maslak V, Savić V, Vasiljević B, O'Connor KE, Nikodinović-Runić J. Highly efficient Michael-type addition of acetaldehyde to beta-nitrostyrenes by whole resting cells of Escherichia coli expressing 4-oxalocrotonate tautomerase. in Bioresource Technology. 2013;142:462-468.
doi:10.1016/j.biortech.2013.05.074 .

Narančić, Tanja, Kadivojević, Jelena, Jovanović, Predrag M., Francuski, Đorđe, Bigović, Miljan, Maslak, Veselin, Savić, Vladimir, Vasiljević, Branka, O'Connor, Kevin E., Nikodinović-Runić, Jasmina, "Highly efficient Michael-type addition of acetaldehyde to beta-nitrostyrenes by whole resting cells of Escherichia coli expressing 4-oxalocrotonate tautomerase" in Bioresource Technology, 142 (2013):462-468,
https://doi.org/10.1016/j.biortech.2013.05.074 . .

TY  - JOUR
AU  - Nikodinović-Runić, Jasmina
AU  - Coulombel, Lydie
AU  - Francuski, Đorđe
AU  - Sharma, Narain D.
AU  - Boyd, Derek R.
AU  - Ferrall, Rory Moore O.
AU  - O'Connor, Kevin E.
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1355
AB  - Nine different sulfur-containing compounds were biotransformed to the corresponding sulfoxides by Escherichia coli Bl21(DE3) cells expressing styrene monooxygenase (SMO) from Pseudomonas putida CA-3. Thioanisole was consumed at 83.3 mu moles min(-1) g cell dry weight(-1) resulting mainly in the formation of R-thioanisole sulfoxide with an enantiomeric excess (ee) value of 45 %. The rate of 2-methyl-, 2-chloro- and 2-bromo-thioanisole consumption was 2-fold lower than that of thioanisole. Surprisingly, the 2-methylthioanisole sulfoxide product had the opposite (S) configuration to that of the other 2-substituted thioanisole derivatives and had a higher ee value (84 %). The rate of oxidation of 4-substituted thioanisoles was higher than the corresponding 2-substituted substrates but the ee values of the products were consistently lower (10-23 %). The rate of benzo[b]thiophene and 2-methylbenzo[b]thiophene sulfoxidation was approximately 10-fold lower than that of thioanisole. The ee value of the benzo[b]thiophene sulfoxide could not be determined as the product racemized rapidly. E. coli cells expressing an engineered SMO (SMOeng R3-11) oxidised 2-substituted thioanisoles between 1.8- and 2.8-fold faster compared to cells expressing the wild-type enzyme. SMOeng R3-11 oxidised benzo[b]thiophene and 2-methylbenzo[b]thiophene 10.1 and 5.6 times faster that the wild-type enzyme. The stereospecificity of the reaction catalysed by SMOeng was unchanged from that of the wild type. Using the X-ray crystal structure of the P. putida S12 SMO, it was evident that the entrance of substrates into the SMO active site is limited by the binding pocket bottleneck formed by the side chains of Val-211 and Asn-46 carboxyamide group.
PB  - Springer, New York
T2  - Applied Microbiology and Biotechnology
T1  - The oxidation of alkylaryl sulfides and benzo[b]thiophenes by Escherichia coli cells expressing wild-type and engineered styrene monooxygenase from Pseudomonas putida CA-3
VL  - 97
IS  - 11
SP  - 4849
EP  - 4858
DO  - 10.1007/s00253-012-4332-5
ER  - 

@article{
author = "Nikodinović-Runić, Jasmina and Coulombel, Lydie and Francuski, Đorđe and Sharma, Narain D. and Boyd, Derek R. and Ferrall, Rory Moore O. and O'Connor, Kevin E.",
year = "2013",
abstract = "Nine different sulfur-containing compounds were biotransformed to the corresponding sulfoxides by Escherichia coli Bl21(DE3) cells expressing styrene monooxygenase (SMO) from Pseudomonas putida CA-3. Thioanisole was consumed at 83.3 mu moles min(-1) g cell dry weight(-1) resulting mainly in the formation of R-thioanisole sulfoxide with an enantiomeric excess (ee) value of 45 %. The rate of 2-methyl-, 2-chloro- and 2-bromo-thioanisole consumption was 2-fold lower than that of thioanisole. Surprisingly, the 2-methylthioanisole sulfoxide product had the opposite (S) configuration to that of the other 2-substituted thioanisole derivatives and had a higher ee value (84 %). The rate of oxidation of 4-substituted thioanisoles was higher than the corresponding 2-substituted substrates but the ee values of the products were consistently lower (10-23 %). The rate of benzo[b]thiophene and 2-methylbenzo[b]thiophene sulfoxidation was approximately 10-fold lower than that of thioanisole. The ee value of the benzo[b]thiophene sulfoxide could not be determined as the product racemized rapidly. E. coli cells expressing an engineered SMO (SMOeng R3-11) oxidised 2-substituted thioanisoles between 1.8- and 2.8-fold faster compared to cells expressing the wild-type enzyme. SMOeng R3-11 oxidised benzo[b]thiophene and 2-methylbenzo[b]thiophene 10.1 and 5.6 times faster that the wild-type enzyme. The stereospecificity of the reaction catalysed by SMOeng was unchanged from that of the wild type. Using the X-ray crystal structure of the P. putida S12 SMO, it was evident that the entrance of substrates into the SMO active site is limited by the binding pocket bottleneck formed by the side chains of Val-211 and Asn-46 carboxyamide group.",
publisher = "Springer, New York",
journal = "Applied Microbiology and Biotechnology",
title = "The oxidation of alkylaryl sulfides and benzo[b]thiophenes by Escherichia coli cells expressing wild-type and engineered styrene monooxygenase from Pseudomonas putida CA-3",
volume = "97",
number = "11",
pages = "4849-4858",
doi = "10.1007/s00253-012-4332-5"
}

Nikodinović-Runić, J., Coulombel, L., Francuski, Đ., Sharma, N. D., Boyd, D. R., Ferrall, R. M. O.,& O'Connor, K. E.. (2013). The oxidation of alkylaryl sulfides and benzo[b]thiophenes by Escherichia coli cells expressing wild-type and engineered styrene monooxygenase from Pseudomonas putida CA-3. in Applied Microbiology and Biotechnology
Springer, New York., 97(11), 4849-4858.
https://doi.org/10.1007/s00253-012-4332-5

Nikodinović-Runić J, Coulombel L, Francuski Đ, Sharma ND, Boyd DR, Ferrall RMO, O'Connor KE. The oxidation of alkylaryl sulfides and benzo[b]thiophenes by Escherichia coli cells expressing wild-type and engineered styrene monooxygenase from Pseudomonas putida CA-3. in Applied Microbiology and Biotechnology. 2013;97(11):4849-4858.
doi:10.1007/s00253-012-4332-5 .

Nikodinović-Runić, Jasmina, Coulombel, Lydie, Francuski, Đorđe, Sharma, Narain D., Boyd, Derek R., Ferrall, Rory Moore O., O'Connor, Kevin E., "The oxidation of alkylaryl sulfides and benzo[b]thiophenes by Escherichia coli cells expressing wild-type and engineered styrene monooxygenase from Pseudomonas putida CA-3" in Applied Microbiology and Biotechnology, 97, no. 11 (2013):4849-4858,
https://doi.org/10.1007/s00253-012-4332-5 . .