TY  - JOUR
AU  - Nikodinović-Runić, Jasmina
AU  - Coulombel, Lydie
AU  - Francuski, Djordje
AU  - Sharma, Narain D.
AU  - Boyd, Derek R.
AU  - Ferrall, Rory Moore O.
AU  - O'Connor, Kevin 
PY  - 2013
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/644
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
EP  - 4858
IS  - 11
SP  - 4849
VL  - 97
DO  - 10.1007/s00253-012-4332-5
ER  - 

@article{
author = "Nikodinović-Runić, Jasmina and Coulombel, Lydie and Francuski, Djordje and Sharma, Narain D. and Boyd, Derek R. and Ferrall, Rory Moore O. and O'Connor, Kevin ",
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",
pages = "4858-4849",
number = "11",
volume = "97",
doi = "10.1007/s00253-012-4332-5"
}

Nikodinović-Runić, J., Coulombel, L., Francuski, D., Sharma, N. D., Boyd, D. R., Ferrall, R. M. O.,& O'Connor, K.. (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 D, Sharma ND, Boyd DR, Ferrall RMO, O'Connor K. 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, Djordje, Sharma, Narain D., Boyd, Derek R., Ferrall, Rory Moore O., O'Connor, Kevin , "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 . .

TY  - JOUR
AU  - Boyd, Derek R.
AU  - Sharma, Narain D.
AU  - McMurray, Brian
AU  - Haughey, Simon A.
AU  - Allen, Christopher C. R.
AU  - Hamilton, John T. G.
AU  - McRoberts, W. Colin
AU  - O'Ferrall, Rory A. More
AU  - Nikodinović-Runić, Jasmina
AU  - Coulombel, Lydie A.
AU  - O'Connor, Kevin 
PY  - 2012
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/612
AB  - Asymmetric heteroatom oxidation of benzo[b]thiophenes to yield the corresponding sulfoxides was catalysed by toluene dioxygenase (TDO), naphthalene dioxygenase (NDO) and styrene monooxygenase (SMO) enzymes present in P. putida mutant and E. coli recombinant whole cells. TDO-catalysed oxidation yielded the relatively unstable benzo[b] thiophene sulfoxide; its dimerization, followed by dehydrogenation, resulted in the isolation of stable tetracyclic sulfoxides as minor products with cis-dihydrodiols being the dominant metabolites. SMO mainly catalysed the formation of enantioenriched benzo[b] thiophene sulfoxide and 2-methyl benzo[b] thiophene sulfoxides which racemized at ambient temperature. The barriers to pyramidal sulfur inversion of 2- and 3-methyl benzo[b] thiophene sulfoxide metabolites, obtained using TDO and NDO as biocatalysts, were found to be ca.: 25-27 kcal mol(-1). The absolute configurations of the benzo[b] thiophene sulfoxides were determined by ECD spectroscopy, X-ray crystallography and stereochemical correlation. A site-directed mutant E. coli strain containing an engineered form of NDO, was found to change the regioselectivity toward preferential oxidation of the thiophene ring rather than the benzene ring.
PB  - Royal Soc Chemistry, Cambridge
T2  - Organic & Biomolecular Chemistry
T1  - Bacterial dioxygenase- and monooxygenase-catalysed sulfoxidation of benzo[b]thiophenes
EP  - 790
IS  - 4
SP  - 782
VL  - 10
DO  - 10.1039/c1ob06678a
ER  - 

@article{
author = "Boyd, Derek R. and Sharma, Narain D. and McMurray, Brian and Haughey, Simon A. and Allen, Christopher C. R. and Hamilton, John T. G. and McRoberts, W. Colin and O'Ferrall, Rory A. More and Nikodinović-Runić, Jasmina and Coulombel, Lydie A. and O'Connor, Kevin ",
year = "2012",
abstract = "Asymmetric heteroatom oxidation of benzo[b]thiophenes to yield the corresponding sulfoxides was catalysed by toluene dioxygenase (TDO), naphthalene dioxygenase (NDO) and styrene monooxygenase (SMO) enzymes present in P. putida mutant and E. coli recombinant whole cells. TDO-catalysed oxidation yielded the relatively unstable benzo[b] thiophene sulfoxide; its dimerization, followed by dehydrogenation, resulted in the isolation of stable tetracyclic sulfoxides as minor products with cis-dihydrodiols being the dominant metabolites. SMO mainly catalysed the formation of enantioenriched benzo[b] thiophene sulfoxide and 2-methyl benzo[b] thiophene sulfoxides which racemized at ambient temperature. The barriers to pyramidal sulfur inversion of 2- and 3-methyl benzo[b] thiophene sulfoxide metabolites, obtained using TDO and NDO as biocatalysts, were found to be ca.: 25-27 kcal mol(-1). The absolute configurations of the benzo[b] thiophene sulfoxides were determined by ECD spectroscopy, X-ray crystallography and stereochemical correlation. A site-directed mutant E. coli strain containing an engineered form of NDO, was found to change the regioselectivity toward preferential oxidation of the thiophene ring rather than the benzene ring.",
publisher = "Royal Soc Chemistry, Cambridge",
journal = "Organic & Biomolecular Chemistry",
title = "Bacterial dioxygenase- and monooxygenase-catalysed sulfoxidation of benzo[b]thiophenes",
pages = "790-782",
number = "4",
volume = "10",
doi = "10.1039/c1ob06678a"
}

Boyd, D. R., Sharma, N. D., McMurray, B., Haughey, S. A., Allen, C. C. R., Hamilton, J. T. G., McRoberts, W. C., O'Ferrall, R. A. M., Nikodinović-Runić, J., Coulombel, L. A.,& O'Connor, K.. (2012). Bacterial dioxygenase- and monooxygenase-catalysed sulfoxidation of benzo[b]thiophenes. in Organic & Biomolecular Chemistry
Royal Soc Chemistry, Cambridge., 10(4), 782-790.
https://doi.org/10.1039/c1ob06678a

Boyd DR, Sharma ND, McMurray B, Haughey SA, Allen CCR, Hamilton JTG, McRoberts WC, O'Ferrall RAM, Nikodinović-Runić J, Coulombel LA, O'Connor K. Bacterial dioxygenase- and monooxygenase-catalysed sulfoxidation of benzo[b]thiophenes. in Organic & Biomolecular Chemistry. 2012;10(4):782-790.
doi:10.1039/c1ob06678a .

Boyd, Derek R., Sharma, Narain D., McMurray, Brian, Haughey, Simon A., Allen, Christopher C. R., Hamilton, John T. G., McRoberts, W. Colin, O'Ferrall, Rory A. More, Nikodinović-Runić, Jasmina, Coulombel, Lydie A., O'Connor, Kevin , "Bacterial dioxygenase- and monooxygenase-catalysed sulfoxidation of benzo[b]thiophenes" in Organic & Biomolecular Chemistry, 10, no. 4 (2012):782-790,
https://doi.org/10.1039/c1ob06678a . .

TY  - JOUR
AU  - Coulombel, Lydie
AU  - Nolan, Louise C.
AU  - Nikodinović-Runić, Jasmina
AU  - Doyle, Evelyn M.
AU  - O'Connor, Kevin 
PY  - 2011
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/503
AB  - Escherichia coli cells, expressing 4-hydroxyphenylacetate 3-hydroxylase, fully transformed 4-halogenated phenols to their equivalent catechols as single products in shaken flasks. 4-Fluorophenol was transformed at a rate 1.6, 1.8, and 3.4-fold higher than the biotransformation of 4-chloro-, 4-bromo-, and 4-iodo- phenol, respectively. A scale-up from shaken flask to a 5 L stirred tank bioreactor was undertaken to develop a bioprocess for the production of 4-substituted halocatechols at higher concentrations and scale. In a stirred tank reactor, the optimized conditions for induction of 4-HPA hydroxylase expression were at 37 C for 3 h. The rate of biotransformation of 4-fluorophenol to 4-fluorocatechol by stirred tank bioreactor grown cells was the same at 1 and 4.8 mM (5.13 mu mol/min/g CDW) once the ratio of biocatalyst (E. coli CDW) to substrate concentration (mM) was maintained at 2:1. At 10.8 mM 4-fluorophenol, the rate of 4-fluorocatechol formation decreased by 4.7-fold. However, the complete transformation of 1.3 g of 4-fluorophenol (10.8 mM) to 4-fluorocatechol was achieved within 7 h in a 1 L reaction volume. Similar to 4-fluorophenol, other 4-substituted halophenols were completely transformed to 4-halocatechols at 2 mM within a 1-2 h period. An increase in 4-halophenol concentration to 4.8 mM resulted in a 2.5-20-fold decrease in biotransformation efficiency depending on the substrate tested. Organic solvent extraction of the 4-halocatechol products followed by column chromatography resulted in the production of purified products with a final yield of between 33% and 38%.
PB  - Springer, New York
T2  - Applied Microbiology and Biotechnology
T1  - Biotransformation of 4-halophenols to 4-halocatechols using Escherichia coli expressing 4-hydroxyphenylacetate 3-hydroxylase
EP  - 1875
IS  - 6
SP  - 1867
VL  - 89
DO  - 10.1007/s00253-010-2969-5
ER  - 

@article{
author = "Coulombel, Lydie and Nolan, Louise C. and Nikodinović-Runić, Jasmina and Doyle, Evelyn M. and O'Connor, Kevin ",
year = "2011",
abstract = "Escherichia coli cells, expressing 4-hydroxyphenylacetate 3-hydroxylase, fully transformed 4-halogenated phenols to their equivalent catechols as single products in shaken flasks. 4-Fluorophenol was transformed at a rate 1.6, 1.8, and 3.4-fold higher than the biotransformation of 4-chloro-, 4-bromo-, and 4-iodo- phenol, respectively. A scale-up from shaken flask to a 5 L stirred tank bioreactor was undertaken to develop a bioprocess for the production of 4-substituted halocatechols at higher concentrations and scale. In a stirred tank reactor, the optimized conditions for induction of 4-HPA hydroxylase expression were at 37 C for 3 h. The rate of biotransformation of 4-fluorophenol to 4-fluorocatechol by stirred tank bioreactor grown cells was the same at 1 and 4.8 mM (5.13 mu mol/min/g CDW) once the ratio of biocatalyst (E. coli CDW) to substrate concentration (mM) was maintained at 2:1. At 10.8 mM 4-fluorophenol, the rate of 4-fluorocatechol formation decreased by 4.7-fold. However, the complete transformation of 1.3 g of 4-fluorophenol (10.8 mM) to 4-fluorocatechol was achieved within 7 h in a 1 L reaction volume. Similar to 4-fluorophenol, other 4-substituted halophenols were completely transformed to 4-halocatechols at 2 mM within a 1-2 h period. An increase in 4-halophenol concentration to 4.8 mM resulted in a 2.5-20-fold decrease in biotransformation efficiency depending on the substrate tested. Organic solvent extraction of the 4-halocatechol products followed by column chromatography resulted in the production of purified products with a final yield of between 33% and 38%.",
publisher = "Springer, New York",
journal = "Applied Microbiology and Biotechnology",
title = "Biotransformation of 4-halophenols to 4-halocatechols using Escherichia coli expressing 4-hydroxyphenylacetate 3-hydroxylase",
pages = "1875-1867",
number = "6",
volume = "89",
doi = "10.1007/s00253-010-2969-5"
}

Coulombel, L., Nolan, L. C., Nikodinović-Runić, J., Doyle, E. M.,& O'Connor, K.. (2011). Biotransformation of 4-halophenols to 4-halocatechols using Escherichia coli expressing 4-hydroxyphenylacetate 3-hydroxylase. in Applied Microbiology and Biotechnology
Springer, New York., 89(6), 1867-1875.
https://doi.org/10.1007/s00253-010-2969-5

Coulombel L, Nolan LC, Nikodinović-Runić J, Doyle EM, O'Connor K. Biotransformation of 4-halophenols to 4-halocatechols using Escherichia coli expressing 4-hydroxyphenylacetate 3-hydroxylase. in Applied Microbiology and Biotechnology. 2011;89(6):1867-1875.
doi:10.1007/s00253-010-2969-5 .

Coulombel, Lydie, Nolan, Louise C., Nikodinović-Runić, Jasmina, Doyle, Evelyn M., O'Connor, Kevin , "Biotransformation of 4-halophenols to 4-halocatechols using Escherichia coli expressing 4-hydroxyphenylacetate 3-hydroxylase" in Applied Microbiology and Biotechnology, 89, no. 6 (2011):1867-1875,
https://doi.org/10.1007/s00253-010-2969-5 . .

TY  - JOUR
AU  - Gursky, Lucas J.
AU  - Nikodinović-Runić, Jasmina
AU  - Feenstra, K. Anton
AU  - O'Connor, Kevin 
PY  - 2010
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/458
AB  - The styAB genes from Pseudomonas putida CA-3, which encode styrene monooxygenase, were subjected to three rounds of in vitro evolution using error-prone polymerase chain reaction with a view to improving the rate of styrene oxide and indene oxide formation. Improvements in styrene monooxygenase activity were monitored using an indole to indigo conversion assay. Each round of random mutagenesis generated variants improved in indigo formation with third round variants improved nine- to 12-fold over the wild type enzyme. Each round of in vitro evolution resulted in two to three amino acid substitutions in styrene monooxygenase. While the majority of mutations occurred in styA (oxygenase), mutations were also observed in styB (reductase). A mutation resulting in the substitution of valine with isoleucine at amino acid residue 303 occurred near the styrene and flavin adenine dinucleotide binding site of styrene monooxygenase. One mutation caused a shift in the reading frame in styA and resulted in a StyA variant that is 19 amino acids longer than the wild-type protein. Whole cells expressing the best styrene monooxygenase variants (round 3) exhibited eight- and 12-fold improvements in styrene and indene oxidation rates compared to the wild-type enzyme. In all cases, a single enantiomer, (S)-styrene oxide, was formed from styrene while (1S,2R)-indene oxide was the predominant enantiomer (e.e. 97%) formed from indene. The average yield of styrene oxide and indene oxide from their respective alkene substrates was 65% and 90%, respectively.
PB  - Springer, New York
T2  - Applied Microbiology and Biotechnology
T1  - In vitro evolution of styrene monooxygenase from Pseudomonas putida CA-3 for improved epoxide synthesis
EP  - 1004
IS  - 4
SP  - 995
VL  - 85
DO  - 10.1007/s00253-009-2096-3
ER  - 

@article{
author = "Gursky, Lucas J. and Nikodinović-Runić, Jasmina and Feenstra, K. Anton and O'Connor, Kevin ",
year = "2010",
abstract = "The styAB genes from Pseudomonas putida CA-3, which encode styrene monooxygenase, were subjected to three rounds of in vitro evolution using error-prone polymerase chain reaction with a view to improving the rate of styrene oxide and indene oxide formation. Improvements in styrene monooxygenase activity were monitored using an indole to indigo conversion assay. Each round of random mutagenesis generated variants improved in indigo formation with third round variants improved nine- to 12-fold over the wild type enzyme. Each round of in vitro evolution resulted in two to three amino acid substitutions in styrene monooxygenase. While the majority of mutations occurred in styA (oxygenase), mutations were also observed in styB (reductase). A mutation resulting in the substitution of valine with isoleucine at amino acid residue 303 occurred near the styrene and flavin adenine dinucleotide binding site of styrene monooxygenase. One mutation caused a shift in the reading frame in styA and resulted in a StyA variant that is 19 amino acids longer than the wild-type protein. Whole cells expressing the best styrene monooxygenase variants (round 3) exhibited eight- and 12-fold improvements in styrene and indene oxidation rates compared to the wild-type enzyme. In all cases, a single enantiomer, (S)-styrene oxide, was formed from styrene while (1S,2R)-indene oxide was the predominant enantiomer (e.e. 97%) formed from indene. The average yield of styrene oxide and indene oxide from their respective alkene substrates was 65% and 90%, respectively.",
publisher = "Springer, New York",
journal = "Applied Microbiology and Biotechnology",
title = "In vitro evolution of styrene monooxygenase from Pseudomonas putida CA-3 for improved epoxide synthesis",
pages = "1004-995",
number = "4",
volume = "85",
doi = "10.1007/s00253-009-2096-3"
}

Gursky, L. J., Nikodinović-Runić, J., Feenstra, K. A.,& O'Connor, K.. (2010). In vitro evolution of styrene monooxygenase from Pseudomonas putida CA-3 for improved epoxide synthesis. in Applied Microbiology and Biotechnology
Springer, New York., 85(4), 995-1004.
https://doi.org/10.1007/s00253-009-2096-3

Gursky LJ, Nikodinović-Runić J, Feenstra KA, O'Connor K. In vitro evolution of styrene monooxygenase from Pseudomonas putida CA-3 for improved epoxide synthesis. in Applied Microbiology and Biotechnology. 2010;85(4):995-1004.
doi:10.1007/s00253-009-2096-3 .

Gursky, Lucas J., Nikodinović-Runić, Jasmina, Feenstra, K. Anton, O'Connor, Kevin , "In vitro evolution of styrene monooxygenase from Pseudomonas putida CA-3 for improved epoxide synthesis" in Applied Microbiology and Biotechnology, 85, no. 4 (2010):995-1004,
https://doi.org/10.1007/s00253-009-2096-3 . .