TY  - JOUR
AU  - Molloy, Susan
AU  - Nikodinović-Runić, Jasmina
AU  - Martin, Leona B.
AU  - Hartmann, Hermann
AU  - Solano, Francisco
AU  - Decker, Heinz
AU  - O'Connor, Kevin E.
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3491
AB  - The tyrosinase gene from Ralstonia solanacearum (GenBank NP518458) was subjected to random mutagenesis resulting in tyrosinase variants (RVC10 and RV145) with up to 3.2-fold improvement in kcat, 5.2-fold lower Km and 16-fold improvement in catalytic efficiency for D-tyrosine. Based on RVC10 and RV145 mutated sequences, single mutation variants were generated with all variants showing increased kcat for D-tyrosine compared to the wild type (WT). All single mutation variants based on RV145 had a higher kcat and Km value compared to the RV145 and thus the combination of four mutations in RV145 was antagonistic for turnover, but synergistic for affinity of the enzyme for D-tyrosine. Single mutation variant 145_V153A exhibited the highest (6.9-fold) improvement in kcat and a 2.4-fold increase in Km compared to the WT. Two single mutation variants, C10_N322S and C10_T183I reduced the Km up to 2.6-fold for D-tyrosine but one variant 145_V153A increased the Km 2.4-fold compared to the WT. Homology based modeling of R. solanacearum tyrosinase showed that mutation V153A disrupts the van der Waals interactions with an -helix providing one of the conserved histidine residues of the active site. The kcat and Km values for L-tyrosine decreased for RV145 and RVC10 compared to the WT. RV145 exhibited a 2.1-fold high catalytic efficiency compared to the WT which is a 7.6-fold lower improvement compared to D-tyrosine. RV145 exhibited a threefold higher monophenolase:diphenolase activity ratio for D-tyrosine:D-DOPA and a 1.4-fold higher L-tyrosine:L-DOPA activity ratio compared to the WT. Biotechnol. Bioeng. 2013; 110: 1849-1857.
PB  - Wiley-Blackwell, Hoboken
T2  - Biotechnology and Bioengineering
T1  - Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches
VL  - 110
IS  - 7
SP  - 1849
EP  - 1857
DO  - 10.1002/bit.24859
ER  - 

@article{
author = "Molloy, Susan and Nikodinović-Runić, Jasmina and Martin, Leona B. and Hartmann, Hermann and Solano, Francisco and Decker, Heinz and O'Connor, Kevin E.",
year = "2013",
abstract = "The tyrosinase gene from Ralstonia solanacearum (GenBank NP518458) was subjected to random mutagenesis resulting in tyrosinase variants (RVC10 and RV145) with up to 3.2-fold improvement in kcat, 5.2-fold lower Km and 16-fold improvement in catalytic efficiency for D-tyrosine. Based on RVC10 and RV145 mutated sequences, single mutation variants were generated with all variants showing increased kcat for D-tyrosine compared to the wild type (WT). All single mutation variants based on RV145 had a higher kcat and Km value compared to the RV145 and thus the combination of four mutations in RV145 was antagonistic for turnover, but synergistic for affinity of the enzyme for D-tyrosine. Single mutation variant 145_V153A exhibited the highest (6.9-fold) improvement in kcat and a 2.4-fold increase in Km compared to the WT. Two single mutation variants, C10_N322S and C10_T183I reduced the Km up to 2.6-fold for D-tyrosine but one variant 145_V153A increased the Km 2.4-fold compared to the WT. Homology based modeling of R. solanacearum tyrosinase showed that mutation V153A disrupts the van der Waals interactions with an -helix providing one of the conserved histidine residues of the active site. The kcat and Km values for L-tyrosine decreased for RV145 and RVC10 compared to the WT. RV145 exhibited a 2.1-fold high catalytic efficiency compared to the WT which is a 7.6-fold lower improvement compared to D-tyrosine. RV145 exhibited a threefold higher monophenolase:diphenolase activity ratio for D-tyrosine:D-DOPA and a 1.4-fold higher L-tyrosine:L-DOPA activity ratio compared to the WT. Biotechnol. Bioeng. 2013; 110: 1849-1857.",
publisher = "Wiley-Blackwell, Hoboken",
journal = "Biotechnology and Bioengineering",
title = "Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches",
volume = "110",
number = "7",
pages = "1849-1857",
doi = "10.1002/bit.24859"
}

Molloy, S., Nikodinović-Runić, J., Martin, L. B., Hartmann, H., Solano, F., Decker, H.,& O'Connor, K. E.. (2013). Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches. in Biotechnology and Bioengineering
Wiley-Blackwell, Hoboken., 110(7), 1849-1857.
https://doi.org/10.1002/bit.24859

Molloy S, Nikodinović-Runić J, Martin LB, Hartmann H, Solano F, Decker H, O'Connor KE. Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches. in Biotechnology and Bioengineering. 2013;110(7):1849-1857.
doi:10.1002/bit.24859 .

Molloy, Susan, Nikodinović-Runić, Jasmina, Martin, Leona B., Hartmann, Hermann, Solano, Francisco, Decker, Heinz, O'Connor, Kevin E., "Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches" in Biotechnology and Bioengineering, 110, no. 7 (2013):1849-1857,
https://doi.org/10.1002/bit.24859 . .

TY  - DATA
AU  - Molloy, Susan
AU  - Nikodinović-Runić, Jasmina
AU  - Martin, Leona B.
AU  - Hartmann, Hermann
AU  - Solano, Francisco
AU  - Decker, Heinz
AU  - O'Connor, Kevin E.
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3492
PB  - Wiley-Blackwell, Hoboken
T2  - Biotechnology and Bioengineering
T1  - Supplementary data for article: Molloy, S.; Nikodinović-Runić, J.; Martin, L. B.; Hartmann, H.; Solano, F.; Decker, H.; O’Connor, K. E. Engineering of a Bacterial Tyrosinase for Improved Catalytic Efficiency towards D-Tyrosine Using Random and Site Directed Mutagenesis Approaches. Biotechnology and Bioengineering 2013, 110 (7), 1849–1857. https://doi.org/10.1002/bit.24859
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3492
ER  - 

@misc{
author = "Molloy, Susan and Nikodinović-Runić, Jasmina and Martin, Leona B. and Hartmann, Hermann and Solano, Francisco and Decker, Heinz and O'Connor, Kevin E.",
year = "2013",
publisher = "Wiley-Blackwell, Hoboken",
journal = "Biotechnology and Bioengineering",
title = "Supplementary data for article: Molloy, S.; Nikodinović-Runić, J.; Martin, L. B.; Hartmann, H.; Solano, F.; Decker, H.; O’Connor, K. E. Engineering of a Bacterial Tyrosinase for Improved Catalytic Efficiency towards D-Tyrosine Using Random and Site Directed Mutagenesis Approaches. Biotechnology and Bioengineering 2013, 110 (7), 1849–1857. https://doi.org/10.1002/bit.24859",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3492"
}

Molloy, S., Nikodinović-Runić, J., Martin, L. B., Hartmann, H., Solano, F., Decker, H.,& O'Connor, K. E.. (2013). Supplementary data for article: Molloy, S.; Nikodinović-Runić, J.; Martin, L. B.; Hartmann, H.; Solano, F.; Decker, H.; O’Connor, K. E. Engineering of a Bacterial Tyrosinase for Improved Catalytic Efficiency towards D-Tyrosine Using Random and Site Directed Mutagenesis Approaches. Biotechnology and Bioengineering 2013, 110 (7), 1849–1857. https://doi.org/10.1002/bit.24859. in Biotechnology and Bioengineering
Wiley-Blackwell, Hoboken..
https://hdl.handle.net/21.15107/rcub_cherry_3492

Molloy S, Nikodinović-Runić J, Martin LB, Hartmann H, Solano F, Decker H, O'Connor KE. Supplementary data for article: Molloy, S.; Nikodinović-Runić, J.; Martin, L. B.; Hartmann, H.; Solano, F.; Decker, H.; O’Connor, K. E. Engineering of a Bacterial Tyrosinase for Improved Catalytic Efficiency towards D-Tyrosine Using Random and Site Directed Mutagenesis Approaches. Biotechnology and Bioengineering 2013, 110 (7), 1849–1857. https://doi.org/10.1002/bit.24859. in Biotechnology and Bioengineering. 2013;.
https://hdl.handle.net/21.15107/rcub_cherry_3492 .

Molloy, Susan, Nikodinović-Runić, Jasmina, Martin, Leona B., Hartmann, Hermann, Solano, Francisco, Decker, Heinz, O'Connor, Kevin E., "Supplementary data for article: Molloy, S.; Nikodinović-Runić, J.; Martin, L. B.; Hartmann, H.; Solano, F.; Decker, H.; O’Connor, K. E. Engineering of a Bacterial Tyrosinase for Improved Catalytic Efficiency towards D-Tyrosine Using Random and Site Directed Mutagenesis Approaches. Biotechnology and Bioengineering 2013, 110 (7), 1849–1857. https://doi.org/10.1002/bit.24859" in Biotechnology and Bioengineering (2013),
https://hdl.handle.net/21.15107/rcub_cherry_3492 .

TY  - JOUR
AU  - Molloy, Susan
AU  - Nikodinović-Runić, Jasmina
AU  - Martin, Leona B.
AU  - Hartmann, Hermann
AU  - Solano, Francisco
AU  - Decker, Heinz
AU  - O'Connor, Kevin E.
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1359
AB  - The tyrosinase gene from Ralstonia solanacearum (GenBank NP518458) was subjected to random mutagenesis resulting in tyrosinase variants (RVC10 and RV145) with up to 3.2-fold improvement in kcat, 5.2-fold lower Km and 16-fold improvement in catalytic efficiency for D-tyrosine. Based on RVC10 and RV145 mutated sequences, single mutation variants were generated with all variants showing increased kcat for D-tyrosine compared to the wild type (WT). All single mutation variants based on RV145 had a higher kcat and Km value compared to the RV145 and thus the combination of four mutations in RV145 was antagonistic for turnover, but synergistic for affinity of the enzyme for D-tyrosine. Single mutation variant 145_V153A exhibited the highest (6.9-fold) improvement in kcat and a 2.4-fold increase in Km compared to the WT. Two single mutation variants, C10_N322S and C10_T183I reduced the Km up to 2.6-fold for D-tyrosine but one variant 145_V153A increased the Km 2.4-fold compared to the WT. Homology based modeling of R. solanacearum tyrosinase showed that mutation V153A disrupts the van der Waals interactions with an -helix providing one of the conserved histidine residues of the active site. The kcat and Km values for L-tyrosine decreased for RV145 and RVC10 compared to the WT. RV145 exhibited a 2.1-fold high catalytic efficiency compared to the WT which is a 7.6-fold lower improvement compared to D-tyrosine. RV145 exhibited a threefold higher monophenolase:diphenolase activity ratio for D-tyrosine:D-DOPA and a 1.4-fold higher L-tyrosine:L-DOPA activity ratio compared to the WT. Biotechnol. Bioeng. 2013; 110: 1849-1857.
PB  - Wiley-Blackwell, Hoboken
T2  - Biotechnology and Bioengineering
T1  - Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches
VL  - 110
IS  - 7
SP  - 1849
EP  - 1857
DO  - 10.1002/bit.24859
ER  - 

@article{
author = "Molloy, Susan and Nikodinović-Runić, Jasmina and Martin, Leona B. and Hartmann, Hermann and Solano, Francisco and Decker, Heinz and O'Connor, Kevin E.",
year = "2013",
abstract = "The tyrosinase gene from Ralstonia solanacearum (GenBank NP518458) was subjected to random mutagenesis resulting in tyrosinase variants (RVC10 and RV145) with up to 3.2-fold improvement in kcat, 5.2-fold lower Km and 16-fold improvement in catalytic efficiency for D-tyrosine. Based on RVC10 and RV145 mutated sequences, single mutation variants were generated with all variants showing increased kcat for D-tyrosine compared to the wild type (WT). All single mutation variants based on RV145 had a higher kcat and Km value compared to the RV145 and thus the combination of four mutations in RV145 was antagonistic for turnover, but synergistic for affinity of the enzyme for D-tyrosine. Single mutation variant 145_V153A exhibited the highest (6.9-fold) improvement in kcat and a 2.4-fold increase in Km compared to the WT. Two single mutation variants, C10_N322S and C10_T183I reduced the Km up to 2.6-fold for D-tyrosine but one variant 145_V153A increased the Km 2.4-fold compared to the WT. Homology based modeling of R. solanacearum tyrosinase showed that mutation V153A disrupts the van der Waals interactions with an -helix providing one of the conserved histidine residues of the active site. The kcat and Km values for L-tyrosine decreased for RV145 and RVC10 compared to the WT. RV145 exhibited a 2.1-fold high catalytic efficiency compared to the WT which is a 7.6-fold lower improvement compared to D-tyrosine. RV145 exhibited a threefold higher monophenolase:diphenolase activity ratio for D-tyrosine:D-DOPA and a 1.4-fold higher L-tyrosine:L-DOPA activity ratio compared to the WT. Biotechnol. Bioeng. 2013; 110: 1849-1857.",
publisher = "Wiley-Blackwell, Hoboken",
journal = "Biotechnology and Bioengineering",
title = "Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches",
volume = "110",
number = "7",
pages = "1849-1857",
doi = "10.1002/bit.24859"
}

Molloy, S., Nikodinović-Runić, J., Martin, L. B., Hartmann, H., Solano, F., Decker, H.,& O'Connor, K. E.. (2013). Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches. in Biotechnology and Bioengineering
Wiley-Blackwell, Hoboken., 110(7), 1849-1857.
https://doi.org/10.1002/bit.24859

Molloy S, Nikodinović-Runić J, Martin LB, Hartmann H, Solano F, Decker H, O'Connor KE. Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches. in Biotechnology and Bioengineering. 2013;110(7):1849-1857.
doi:10.1002/bit.24859 .

Molloy, Susan, Nikodinović-Runić, Jasmina, Martin, Leona B., Hartmann, Hermann, Solano, Francisco, Decker, Heinz, O'Connor, Kevin E., "Engineering of a bacterial tyrosinase for improved catalytic efficiency towards D-tyrosine using random and site directed mutagenesis approaches" in Biotechnology and Bioengineering, 110, no. 7 (2013):1849-1857,
https://doi.org/10.1002/bit.24859 . .