New Labeled PET Analogues Enable the Functional Screening and Characterization of PET-Degrading Enzymes
Само за регистроване кориснике
2024
Аутори
Taxeidis, GeorgeĐapović, Milica
Nikolaivits, Efstratios
Maslak, Veselin
Nikodinović-Runić, Jasmina
Topakas, Evangelos
Чланак у часопису (Рецензирана верзија)
Метаподаци
Приказ свих података о документуАпстракт
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, offerin...g 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.
Кључне речи:
polyethylene terephthalate / PETase / plastic degradation / screening / fluorescent substrates / kineticsИзвор:
ACS Sustainable Chemistry & Engineering, 2024Издавач:
- American Chemical Society
Напомена:
- The Supporting Information https://pubs.acs.org/doi/10.1021/acssuschemeng.4c00143
- This is the peer reviewed version of the paper: Taxeidis, G., Djapovic, M., Nikolaivits, E., Maslak, V., Nikodinovic-Runic, J., & Topakas, E. (2024). New Labeled PET Analogues Enable the Functional Screening and Characterization of PET-Degrading Enzymes. ACS Sustainable Chemistry & Engineering. https://doi.org/10.1021/acssuschemeng.4c00143
URI
https://doi.org/10.1021/acssuschemeng.4c00143https://imagine.imgge.bg.ac.rs/handle/123456789/2339
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - 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 . .