TY  - JOUR
AU  - Taxeidis, George
AU  - Đapović, Milica
AU  - Nikolaivits, Efstratios
AU  - Maslak, Veselin
AU  - Nikodinović-Runić, Jasmina
AU  - Topakas, Evangelos
PY  - 2024
UR  - https://doi.org/10.1021/acssuschemeng.4c00143
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2339
AB  - The discovery and engineering of novel biocatalysts capable of depolymerizing polyethylene terephthalate (PET) have gained significant attention since the need for green technologies to combat plastic pollution has become increasingly urgent. This study focuses on the development of novel substrates that can indicate enzymes with PET hydrolytic activity, streamlining the process of enzyme evaluation and selection. Four novel substrates, mimicking the structure of PET, were chemically synthesized and labeled with fluorogenic or chromogenic moieties, enabling the direct analysis of candidate enzymes without complex preparatory or analysis steps. The fluorogenic substrates, mUPET1, mUPET2, and mUPET3, not only identify enzymes capable of PET breakdown but also differentiate those with exceptional performance on the polymer, such as the benchmark PETase, LCCICCG. Among the substrates, the chromogenic p-NPhPET3 stands out as a reliable tool for screening both pure and crude enzymes, offering advantages over fluorogenic substrates such as ease of assay using UV–vis spectroscopy and compatibility with crude enzyme samples. However, ferulic acid esterases and mono-(2-hydroxyethyl) terephthalate esterases (MHETases), which exhibit remarkably high affinity for PET oligomers, also show high catalytic activity on these substrates. The substrates introduced in this study hold significant value in the function-based screening and characterization of enzymes that degrade PET, as well as the the potential to be used in screening mutant libraries derived from directed evolution experiments. Following this approach, a rapid and dependable assay method can be carried out using basic laboratory infrastructure, eliminating the necessity for intricate preparatory procedures before analysis.
PB  - American Chemical Society
T2  - ACS Sustainable Chemistry & Engineering
T1  - New Labeled PET Analogues Enable the Functional Screening and Characterization of PET-Degrading Enzymes
DO  - 10.1021/acssuschemeng.4c00143
ER  - 

@article{
author = "Taxeidis, George and Đapović, Milica and Nikolaivits, Efstratios and Maslak, Veselin and Nikodinović-Runić, Jasmina and Topakas, Evangelos",
year = "2024",
abstract = "The discovery and engineering of novel biocatalysts capable of depolymerizing polyethylene terephthalate (PET) have gained significant attention since the need for green technologies to combat plastic pollution has become increasingly urgent. This study focuses on the development of novel substrates that can indicate enzymes with PET hydrolytic activity, streamlining the process of enzyme evaluation and selection. Four novel substrates, mimicking the structure of PET, were chemically synthesized and labeled with fluorogenic or chromogenic moieties, enabling the direct analysis of candidate enzymes without complex preparatory or analysis steps. The fluorogenic substrates, mUPET1, mUPET2, and mUPET3, not only identify enzymes capable of PET breakdown but also differentiate those with exceptional performance on the polymer, such as the benchmark PETase, LCCICCG. Among the substrates, the chromogenic p-NPhPET3 stands out as a reliable tool for screening both pure and crude enzymes, offering advantages over fluorogenic substrates such as ease of assay using UV–vis spectroscopy and compatibility with crude enzyme samples. However, ferulic acid esterases and mono-(2-hydroxyethyl) terephthalate esterases (MHETases), which exhibit remarkably high affinity for PET oligomers, also show high catalytic activity on these substrates. The substrates introduced in this study hold significant value in the function-based screening and characterization of enzymes that degrade PET, as well as the the potential to be used in screening mutant libraries derived from directed evolution experiments. Following this approach, a rapid and dependable assay method can be carried out using basic laboratory infrastructure, eliminating the necessity for intricate preparatory procedures before analysis.",
publisher = "American Chemical Society",
journal = "ACS Sustainable Chemistry & Engineering",
title = "New Labeled PET Analogues Enable the Functional Screening and Characterization of PET-Degrading Enzymes",
doi = "10.1021/acssuschemeng.4c00143"
}

Taxeidis, G., Đapović, M., Nikolaivits, E., Maslak, V., Nikodinović-Runić, J.,& Topakas, E.. (2024). New Labeled PET Analogues Enable the Functional Screening and Characterization of PET-Degrading Enzymes. in ACS Sustainable Chemistry & Engineering
American Chemical Society..
https://doi.org/10.1021/acssuschemeng.4c00143

Taxeidis G, Đapović M, Nikolaivits E, Maslak V, Nikodinović-Runić J, Topakas E. New Labeled PET Analogues Enable the Functional Screening and Characterization of PET-Degrading Enzymes. in ACS Sustainable Chemistry & Engineering. 2024;.
doi:10.1021/acssuschemeng.4c00143 .

Taxeidis, George, Đapović, Milica, Nikolaivits, Efstratios, Maslak, Veselin, Nikodinović-Runić, Jasmina, Topakas, Evangelos, "New Labeled PET Analogues Enable the Functional Screening and Characterization of PET-Degrading Enzymes" in ACS Sustainable Chemistry & Engineering (2024),
https://doi.org/10.1021/acssuschemeng.4c00143 . .

TY  - JOUR
AU  - Đapović, Milica
AU  - Milivojević, Dušan
AU  - Ilić-Tomić, Tatjana
AU  - Lješević, Marija
AU  - Nikolaivits, Efstratios
AU  - Topakas, Evangelos
AU  - Maslak, Veselin
AU  - Nikodinović-Runić, Jasmina
PY  - 2021
UR  - https://www.sciencedirect.com/science/article/pii/S0045653521004744
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1732
AB  - Polyethylene terephthalate (PET) is widely used material and as such became highly enriched in nature. It is generally considered inert and safe plastic, but due to the recent increased efforts to break-down PET using biotechnological approaches, we realized the scarcity of information about structural analysis of possible degradation products and their ecotoxicological assessment. Therefore, in this study, 11 compounds belonging to the group of PET precursors and possible degradation products have been comprehensively characterized. Seven of these compounds including 1-(2-hydroxyethyl)-4-methylterephthalate, ethylene glycol bis(methyl terephthalate), methyl bis(2-hydroxyethyl terephtahalate), 1,4-benzenedicarboxylic acid, 1,4-bis[2-[[4-(methoxycarbonyl)benzoyl]oxy]ethyl] ester and methyl tris(2-hydroxyethyl terephthalate) corresponding to mono-, 1.5-, di-, 2,5- and trimer of PET were synthetized and structurally characterized for the first time. In-silico druglikeness and physico-chemical properties of these compounds were predicted using variety of platforms. No antimicrobial properties were detected even at 1000 μg/mL. Ecotoxicological impact of the compounds against marine bacteria Allivibrio fischeri proved that the 6 out of 11 tested PET-associated compounds may be classified as harmful to aquatic microorganisms, with PET trimer being one of the most toxic. In comparison, most of the compounds were not toxic on human lung fibroblasts (MRC-5) at 200 μg/mL with inhibiting concentration (IC50) values of 30 μg/mL and 50 μg/mL determined for PET dimer and trimer. Only three of these compounds including PET monomer were toxic to nematode Caenorhabditis elegans at high concentration of 500 μg/mL. In terms of the applicative potential, PET dimer can be used as suitable substrate for the screening, identification and characterization of novel PET-depolymerizing enzymes.
PB  - Elsevier
T2  - Chemosphere
T2  - ChemosphereChemosphere
T1  - Synthesis and characterization of polyethylene terephthalate (PET) precursors and potential degradation products: Toxicity study and application in discovery of novel PETases
SP  - 130005
VL  - 275
DO  - 10.1016/j.chemosphere.2021.130005
ER  - 

@article{
author = "Đapović, Milica and Milivojević, Dušan and Ilić-Tomić, Tatjana and Lješević, Marija and Nikolaivits, Efstratios and Topakas, Evangelos and Maslak, Veselin and Nikodinović-Runić, Jasmina",
year = "2021",
abstract = "Polyethylene terephthalate (PET) is widely used material and as such became highly enriched in nature. It is generally considered inert and safe plastic, but due to the recent increased efforts to break-down PET using biotechnological approaches, we realized the scarcity of information about structural analysis of possible degradation products and their ecotoxicological assessment. Therefore, in this study, 11 compounds belonging to the group of PET precursors and possible degradation products have been comprehensively characterized. Seven of these compounds including 1-(2-hydroxyethyl)-4-methylterephthalate, ethylene glycol bis(methyl terephthalate), methyl bis(2-hydroxyethyl terephtahalate), 1,4-benzenedicarboxylic acid, 1,4-bis[2-[[4-(methoxycarbonyl)benzoyl]oxy]ethyl] ester and methyl tris(2-hydroxyethyl terephthalate) corresponding to mono-, 1.5-, di-, 2,5- and trimer of PET were synthetized and structurally characterized for the first time. In-silico druglikeness and physico-chemical properties of these compounds were predicted using variety of platforms. No antimicrobial properties were detected even at 1000 μg/mL. Ecotoxicological impact of the compounds against marine bacteria Allivibrio fischeri proved that the 6 out of 11 tested PET-associated compounds may be classified as harmful to aquatic microorganisms, with PET trimer being one of the most toxic. In comparison, most of the compounds were not toxic on human lung fibroblasts (MRC-5) at 200 μg/mL with inhibiting concentration (IC50) values of 30 μg/mL and 50 μg/mL determined for PET dimer and trimer. Only three of these compounds including PET monomer were toxic to nematode Caenorhabditis elegans at high concentration of 500 μg/mL. In terms of the applicative potential, PET dimer can be used as suitable substrate for the screening, identification and characterization of novel PET-depolymerizing enzymes.",
publisher = "Elsevier",
journal = "Chemosphere, ChemosphereChemosphere",
title = "Synthesis and characterization of polyethylene terephthalate (PET) precursors and potential degradation products: Toxicity study and application in discovery of novel PETases",
pages = "130005",
volume = "275",
doi = "10.1016/j.chemosphere.2021.130005"
}

Đapović, M., Milivojević, D., Ilić-Tomić, T., Lješević, M., Nikolaivits, E., Topakas, E., Maslak, V.,& Nikodinović-Runić, J.. (2021). Synthesis and characterization of polyethylene terephthalate (PET) precursors and potential degradation products: Toxicity study and application in discovery of novel PETases. in Chemosphere
Elsevier., 275, 130005.
https://doi.org/10.1016/j.chemosphere.2021.130005

Đapović M, Milivojević D, Ilić-Tomić T, Lješević M, Nikolaivits E, Topakas E, Maslak V, Nikodinović-Runić J. Synthesis and characterization of polyethylene terephthalate (PET) precursors and potential degradation products: Toxicity study and application in discovery of novel PETases. in Chemosphere. 2021;275:130005.
doi:10.1016/j.chemosphere.2021.130005 .

Đapović, Milica, Milivojević, Dušan, Ilić-Tomić, Tatjana, Lješević, Marija, Nikolaivits, Efstratios, Topakas, Evangelos, Maslak, Veselin, Nikodinović-Runić, Jasmina, "Synthesis and characterization of polyethylene terephthalate (PET) precursors and potential degradation products: Toxicity study and application in discovery of novel PETases" in Chemosphere, 275 (2021):130005,
https://doi.org/10.1016/j.chemosphere.2021.130005 . .

TY  - JOUR
AU  - Đapović, Milica
AU  - Milivojević, Dušan
AU  - Ilić-Tomić, Tatjana
AU  - Ljesević, Marija
AU  - Nikolaivits, Efstratios
AU  - Topakas, Evangelos
AU  - Maslak, Veselin
AU  - Nikodinović-Runić, Jasmina
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1415
AB  - Polyethylene terephthalate (PET) is widely used material and as such became highly enriched in nature. It is generally considered inert and safe plastic, but due to the recent increased efforts to break-down PET using biotechnological approaches, we realized the scarcity of information about structural analysis of possible degradation products and their ecotoxicological assessment. Therefore, in this study, 11 compounds belonging to the group of PET precursors and possible degradation products have been comprehensively characterized. Seven of these compounds including 1-(2-hydroxyethyl)-4methylterephthalate, ethylene glycol bis(methyl terephthalate), methyl bis(2-hydroxyethyl terephtahalate), 1,4-benzenedicarboxylic acid, 1,4-bis[2-[[4-(methoxycarbonyl)benzoyl]oxy]ethyl] ester and methyl tris(2-hydroxyethyl terephthalate) corresponding to mono-, 1.5-, di-, 2,5- and trimer of PET were synthetized and structurally characterized for the first time. In-silico druglikeness and physico-chemical properties of these compounds were predicted using variety of platforms. No antimicrobial properties were detected even at 1000 mg/mL. Ecotoxicological impact of the compounds against marine bacteria Allivibrio fischeri proved that the 6 out of 11 tested PET-associated compounds may be classified as harmful to aquatic microorganisms, with PET trimer being one of the most toxic. In comparison, most of the compounds were not toxic on human lung fibroblasts (MRC-5) at 200 mg/mL with inhibiting concentration (IC50) values of 30 mg/mL and 50 mg/mL determined for PET dimer and trimer. Only three of these compounds including PET monomer were toxic to nematode Caenorhabditis elegans at high concentration of 500 mg/mL. In terms of the applicative potential, PET dimer can be used as suitable substrate for the screening, identification and characterization of novel PET-depolymerizing enzymes.
PB  - Pergamon-Elsevier Science Ltd, Oxford
T2  - Chemosphere
T1  - Synthesis and characterization of polyethylene terephthalate (PET) precursors and potential degradation products: Toxicity study and application in discovery of novel PETases
SP  - 130005
VL  - 275
DO  - 10.1016/j.chemosphere.2021.130005
ER  - 

@article{
author = "Đapović, Milica and Milivojević, Dušan and Ilić-Tomić, Tatjana and Ljesević, Marija and Nikolaivits, Efstratios and Topakas, Evangelos and Maslak, Veselin and Nikodinović-Runić, Jasmina",
year = "2021",
abstract = "Polyethylene terephthalate (PET) is widely used material and as such became highly enriched in nature. It is generally considered inert and safe plastic, but due to the recent increased efforts to break-down PET using biotechnological approaches, we realized the scarcity of information about structural analysis of possible degradation products and their ecotoxicological assessment. Therefore, in this study, 11 compounds belonging to the group of PET precursors and possible degradation products have been comprehensively characterized. Seven of these compounds including 1-(2-hydroxyethyl)-4methylterephthalate, ethylene glycol bis(methyl terephthalate), methyl bis(2-hydroxyethyl terephtahalate), 1,4-benzenedicarboxylic acid, 1,4-bis[2-[[4-(methoxycarbonyl)benzoyl]oxy]ethyl] ester and methyl tris(2-hydroxyethyl terephthalate) corresponding to mono-, 1.5-, di-, 2,5- and trimer of PET were synthetized and structurally characterized for the first time. In-silico druglikeness and physico-chemical properties of these compounds were predicted using variety of platforms. No antimicrobial properties were detected even at 1000 mg/mL. Ecotoxicological impact of the compounds against marine bacteria Allivibrio fischeri proved that the 6 out of 11 tested PET-associated compounds may be classified as harmful to aquatic microorganisms, with PET trimer being one of the most toxic. In comparison, most of the compounds were not toxic on human lung fibroblasts (MRC-5) at 200 mg/mL with inhibiting concentration (IC50) values of 30 mg/mL and 50 mg/mL determined for PET dimer and trimer. Only three of these compounds including PET monomer were toxic to nematode Caenorhabditis elegans at high concentration of 500 mg/mL. In terms of the applicative potential, PET dimer can be used as suitable substrate for the screening, identification and characterization of novel PET-depolymerizing enzymes.",
publisher = "Pergamon-Elsevier Science Ltd, Oxford",
journal = "Chemosphere",
title = "Synthesis and characterization of polyethylene terephthalate (PET) precursors and potential degradation products: Toxicity study and application in discovery of novel PETases",
pages = "130005",
volume = "275",
doi = "10.1016/j.chemosphere.2021.130005"
}

Đapović, M., Milivojević, D., Ilić-Tomić, T., Ljesević, M., Nikolaivits, E., Topakas, E., Maslak, V.,& Nikodinović-Runić, J.. (2021). Synthesis and characterization of polyethylene terephthalate (PET) precursors and potential degradation products: Toxicity study and application in discovery of novel PETases. in Chemosphere
Pergamon-Elsevier Science Ltd, Oxford., 275, 130005.
https://doi.org/10.1016/j.chemosphere.2021.130005

Đapović M, Milivojević D, Ilić-Tomić T, Ljesević M, Nikolaivits E, Topakas E, Maslak V, Nikodinović-Runić J. Synthesis and characterization of polyethylene terephthalate (PET) precursors and potential degradation products: Toxicity study and application in discovery of novel PETases. in Chemosphere. 2021;275:130005.
doi:10.1016/j.chemosphere.2021.130005 .

Đapović, Milica, Milivojević, Dušan, Ilić-Tomić, Tatjana, Ljesević, Marija, Nikolaivits, Efstratios, Topakas, Evangelos, Maslak, Veselin, Nikodinović-Runić, Jasmina, "Synthesis and characterization of polyethylene terephthalate (PET) precursors and potential degradation products: Toxicity study and application in discovery of novel PETases" in Chemosphere, 275 (2021):130005,
https://doi.org/10.1016/j.chemosphere.2021.130005 . .

TY  - DATA
AU  - Đapović, Milica
AU  - Milivojević, Dušan
AU  - Ilić-Tomić, Tatjana
AU  - Lješević, Marija
AU  - Nikolaivits, Efstratios
AU  - Topakas, Evangelos
AU  - Maslak, Veselin
AU  - Nikodinović-Runić, Jasmina
PY  - 2021
UR  - https://www.sciencedirect.com/science/article/pii/S0045653521004744
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1733
PB  - Elsevier
T2  - Chemosphere
T1  - Supplementary data for the article: Djapovic, M.; Milivojevic, D.; Ilic-Tomic, T.; Lješević, M.; Nikolaivits, E.; Topakas, E.; Maslak, V.; Nikodinovic-Runic, J. Synthesis and Characterization of Polyethylene Terephthalate (PET) Precursors and Potential Degradation Products: Toxicity Study and Application in Discovery of Novel PETases. Chemosphere 2021, 275, 130005. https://doi.org/10.1016/j.chemosphere.2021.130005.
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1733
ER  - 

@misc{
author = "Đapović, Milica and Milivojević, Dušan and Ilić-Tomić, Tatjana and Lješević, Marija and Nikolaivits, Efstratios and Topakas, Evangelos and Maslak, Veselin and Nikodinović-Runić, Jasmina",
year = "2021",
publisher = "Elsevier",
journal = "Chemosphere",
title = "Supplementary data for the article: Djapovic, M.; Milivojevic, D.; Ilic-Tomic, T.; Lješević, M.; Nikolaivits, E.; Topakas, E.; Maslak, V.; Nikodinovic-Runic, J. Synthesis and Characterization of Polyethylene Terephthalate (PET) Precursors and Potential Degradation Products: Toxicity Study and Application in Discovery of Novel PETases. Chemosphere 2021, 275, 130005. https://doi.org/10.1016/j.chemosphere.2021.130005.",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1733"
}

Đapović, M., Milivojević, D., Ilić-Tomić, T., Lješević, M., Nikolaivits, E., Topakas, E., Maslak, V.,& Nikodinović-Runić, J.. (2021). Supplementary data for the article: Djapovic, M.; Milivojevic, D.; Ilic-Tomic, T.; Lješević, M.; Nikolaivits, E.; Topakas, E.; Maslak, V.; Nikodinovic-Runic, J. Synthesis and Characterization of Polyethylene Terephthalate (PET) Precursors and Potential Degradation Products: Toxicity Study and Application in Discovery of Novel PETases. Chemosphere 2021, 275, 130005. https://doi.org/10.1016/j.chemosphere.2021.130005.. in Chemosphere
Elsevier..
https://hdl.handle.net/21.15107/rcub_imagine_1733

Đapović M, Milivojević D, Ilić-Tomić T, Lješević M, Nikolaivits E, Topakas E, Maslak V, Nikodinović-Runić J. Supplementary data for the article: Djapovic, M.; Milivojevic, D.; Ilic-Tomic, T.; Lješević, M.; Nikolaivits, E.; Topakas, E.; Maslak, V.; Nikodinovic-Runic, J. Synthesis and Characterization of Polyethylene Terephthalate (PET) Precursors and Potential Degradation Products: Toxicity Study and Application in Discovery of Novel PETases. Chemosphere 2021, 275, 130005. https://doi.org/10.1016/j.chemosphere.2021.130005.. in Chemosphere. 2021;.
https://hdl.handle.net/21.15107/rcub_imagine_1733 .

Đapović, Milica, Milivojević, Dušan, Ilić-Tomić, Tatjana, Lješević, Marija, Nikolaivits, Efstratios, Topakas, Evangelos, Maslak, Veselin, Nikodinović-Runić, Jasmina, "Supplementary data for the article: Djapovic, M.; Milivojevic, D.; Ilic-Tomic, T.; Lješević, M.; Nikolaivits, E.; Topakas, E.; Maslak, V.; Nikodinovic-Runic, J. Synthesis and Characterization of Polyethylene Terephthalate (PET) Precursors and Potential Degradation Products: Toxicity Study and Application in Discovery of Novel PETases. Chemosphere 2021, 275, 130005. https://doi.org/10.1016/j.chemosphere.2021.130005." in Chemosphere (2021),
https://hdl.handle.net/21.15107/rcub_imagine_1733 .