Comparative performance of PETase as a function of reaction conditions, substrate properties, and product accumulation
Abstract
Abstract There is keen interest to develop new technologies to recycle the plastic poly(ethylene terephthalate) (PET). To this end, the use of PET‐hydrolyzing enzymes has shown promise for PET deconstruction to its monomers, terephthalate (TPA) and ethylene glycol (EG). Here, the Ideonella sakaiensis PETase wild‐type enzyme was compared to a previously reported improved variant (W159H/S238F). The thermostability of each enzyme was compared and a 1.45 Å resolution structure of the mutant was described, highlighting changes in the substrate binding cleft compared to the wild‐type enzyme. Subsequently, the performance of the wild‐type and variant enzyme was compared as a function of temperature, substrate morphology, and reaction mixture composition. These studies showed that reaction temperature had the strongest influence on performance between the two enzymes. It was also shown that both enzymes achieved higher levels of PET conversion for substrates with moderate crystallinity relative to amorphous substrates. Finally, the impact of product accumulation on reaction progress was assessed for the hydrolysis of both PET and bis(2‐hydroxyethyl) terephthalate (BHET). Each enzyme displayed different inhibition profiles to mono(2‐hydroxyethyl) terephthalate (MHET) and TPA, while both were sensitive to inhibition by EG. Overall, this study highlights the importance of reaction conditions, substrate selection, and product accumulation formore »
- Authors:
-
- National Renewable Energy Lab. (NREL), Golden, CO (United States); BOTTLE Consortium, Golden, CO (United States)
- BOTTLE Consortium, Golden, CO (United States); Univ. of Portsmouth (United Kingdom)
- Univ. of Portsmouth (United Kingdom)
- Publication Date:
- Research Org.:
- National Renewable Energy Laboratory (NREL), Golden, CO (United States)
- Sponsoring Org.:
- USDOE Office of Energy Efficiency and Renewable Energy (EERE), Transportation Office. Bioenergy Technologies Office; Bio-Optimized Technologies to keep Thermoplastics out of Landfills and the Environment (BOTTLE) Consortium; BBSRC; USDOE
- OSTI Identifier:
- 1826114
- Alternate Identifier(s):
- OSTI ID: 1829155
- Report Number(s):
- NREL/JA-2800-81155
Journal ID: ISSN 1864-5631; MainId:79931;UUID:780b5d2f-6edd-460f-b6dd-8338cf4609ad;MainAdminID:63146
- Grant/Contract Number:
- AC36-08GO28308; BB/P011918/1
- Resource Type:
- Accepted Manuscript
- Journal Name:
- ChemSusChem
- Additional Journal Information:
- Journal Volume: 15; Journal Issue: 1; Journal ID: ISSN 1864-5631
- Publisher:
- ChemPubSoc Europe
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; chemical recycling; interfacial biocatalysis; IsPETase; kinetics; Michaelis-Menten; PET hydrolase
Citation Formats
Erickson, Erika, Shakespeare, Thomas J., Bratti, Felicia, Buss, Bonnie L., Graham, Rosie, Hawkins, McKenzie A., Konig, Gerhard, Michener, William E., Miscall, Joel, Ramirez, Kelsey J., Rorrer, Nicholas A., Zahn, Michael, Pickford, Andrew R., McGeehan, John E., and Beckham, Gregg T. Comparative performance of PETase as a function of reaction conditions, substrate properties, and product accumulation. United States: N. p., 2021.
Web. doi:10.1002/cssc.202101932.
Erickson, Erika, Shakespeare, Thomas J., Bratti, Felicia, Buss, Bonnie L., Graham, Rosie, Hawkins, McKenzie A., Konig, Gerhard, Michener, William E., Miscall, Joel, Ramirez, Kelsey J., Rorrer, Nicholas A., Zahn, Michael, Pickford, Andrew R., McGeehan, John E., & Beckham, Gregg T. Comparative performance of PETase as a function of reaction conditions, substrate properties, and product accumulation. United States. https://doi.org/10.1002/cssc.202101932
Erickson, Erika, Shakespeare, Thomas J., Bratti, Felicia, Buss, Bonnie L., Graham, Rosie, Hawkins, McKenzie A., Konig, Gerhard, Michener, William E., Miscall, Joel, Ramirez, Kelsey J., Rorrer, Nicholas A., Zahn, Michael, Pickford, Andrew R., McGeehan, John E., and Beckham, Gregg T. Wed .
"Comparative performance of PETase as a function of reaction conditions, substrate properties, and product accumulation". United States. https://doi.org/10.1002/cssc.202101932. https://www.osti.gov/servlets/purl/1826114.
@article{osti_1826114,
title = {Comparative performance of PETase as a function of reaction conditions, substrate properties, and product accumulation},
author = {Erickson, Erika and Shakespeare, Thomas J. and Bratti, Felicia and Buss, Bonnie L. and Graham, Rosie and Hawkins, McKenzie A. and Konig, Gerhard and Michener, William E. and Miscall, Joel and Ramirez, Kelsey J. and Rorrer, Nicholas A. and Zahn, Michael and Pickford, Andrew R. and McGeehan, John E. and Beckham, Gregg T.},
abstractNote = {Abstract There is keen interest to develop new technologies to recycle the plastic poly(ethylene terephthalate) (PET). To this end, the use of PET‐hydrolyzing enzymes has shown promise for PET deconstruction to its monomers, terephthalate (TPA) and ethylene glycol (EG). Here, the Ideonella sakaiensis PETase wild‐type enzyme was compared to a previously reported improved variant (W159H/S238F). The thermostability of each enzyme was compared and a 1.45 Å resolution structure of the mutant was described, highlighting changes in the substrate binding cleft compared to the wild‐type enzyme. Subsequently, the performance of the wild‐type and variant enzyme was compared as a function of temperature, substrate morphology, and reaction mixture composition. These studies showed that reaction temperature had the strongest influence on performance between the two enzymes. It was also shown that both enzymes achieved higher levels of PET conversion for substrates with moderate crystallinity relative to amorphous substrates. Finally, the impact of product accumulation on reaction progress was assessed for the hydrolysis of both PET and bis(2‐hydroxyethyl) terephthalate (BHET). Each enzyme displayed different inhibition profiles to mono(2‐hydroxyethyl) terephthalate (MHET) and TPA, while both were sensitive to inhibition by EG. Overall, this study highlights the importance of reaction conditions, substrate selection, and product accumulation for catalytic performance of PET‐hydrolyzing enzymes, which have implications for enzyme screening in the development of enzyme‐based polyester recycling.},
doi = {10.1002/cssc.202101932},
journal = {ChemSusChem},
number = 1,
volume = 15,
place = {United States},
year = {Wed Sep 29 00:00:00 EDT 2021},
month = {Wed Sep 29 00:00:00 EDT 2021}
}
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