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Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14

The successful enzymatic degradation of polyester substrates has fueled worldwide investigation into the treatment of plastic waste using bio-based processes. Within this realm, marine-associated microorganisms have emerged as a promising source of polyester-degrading enzymes. In this work, we descr...

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Autores principales: Carr, Clodagh M., Keller, Malene B., Paul, Bijoya, Schubert, Sune W., Clausen, Kristine S. R., Jensen, Kenneth, Clarke, David J., Westh, Peter, Dobson, Alan D. W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10213408/
https://www.ncbi.nlm.nih.gov/pubmed/37250061
http://dx.doi.org/10.3389/fmicb.2023.1170880
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author Carr, Clodagh M.
Keller, Malene B.
Paul, Bijoya
Schubert, Sune W.
Clausen, Kristine S. R.
Jensen, Kenneth
Clarke, David J.
Westh, Peter
Dobson, Alan D. W.
author_facet Carr, Clodagh M.
Keller, Malene B.
Paul, Bijoya
Schubert, Sune W.
Clausen, Kristine S. R.
Jensen, Kenneth
Clarke, David J.
Westh, Peter
Dobson, Alan D. W.
author_sort Carr, Clodagh M.
collection PubMed
description The successful enzymatic degradation of polyester substrates has fueled worldwide investigation into the treatment of plastic waste using bio-based processes. Within this realm, marine-associated microorganisms have emerged as a promising source of polyester-degrading enzymes. In this work, we describe the hydrolysis of the synthetic polymer PET by SM14est, a polyesterase which was previously identified from Streptomyces sp. SM14, an isolate of the marine sponge Haliclona simulans. The PET hydrolase activity of purified SM14est was assessed using a suspension-based assay and subsequent analysis of reaction products by UV-spectrophotometry and RP-HPLC. SM14est displayed a preference for high salt conditions, with activity significantly increasing at sodium chloride concentrations from 100 mM up to 1,000 mM. The initial rate of PET hydrolysis by SM14est was determined to be 0.004 s(−1) at 45°C, which was increased by 5-fold to 0.02 s(−1) upon addition of 500 mM sodium chloride. Sequence alignment and structural comparison with known PET hydrolases, including the marine halophile PET6, and the highly efficient, thermophilic PHL7, revealed conserved features of interest. Based on this work, SM14est emerges as a useful enzyme that is more similar to key players in the area of PET hydrolysis, like PHL7 and IsPETase, than it is to its marine counterparts. Salt-tolerant polyesterases such as SM14est are potentially valuable in the biological degradation of plastic particles that readily contaminate marine ecosystems and industrial wastewaters.
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spelling pubmed-102134082023-05-27 Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14 Carr, Clodagh M. Keller, Malene B. Paul, Bijoya Schubert, Sune W. Clausen, Kristine S. R. Jensen, Kenneth Clarke, David J. Westh, Peter Dobson, Alan D. W. Front Microbiol Microbiology The successful enzymatic degradation of polyester substrates has fueled worldwide investigation into the treatment of plastic waste using bio-based processes. Within this realm, marine-associated microorganisms have emerged as a promising source of polyester-degrading enzymes. In this work, we describe the hydrolysis of the synthetic polymer PET by SM14est, a polyesterase which was previously identified from Streptomyces sp. SM14, an isolate of the marine sponge Haliclona simulans. The PET hydrolase activity of purified SM14est was assessed using a suspension-based assay and subsequent analysis of reaction products by UV-spectrophotometry and RP-HPLC. SM14est displayed a preference for high salt conditions, with activity significantly increasing at sodium chloride concentrations from 100 mM up to 1,000 mM. The initial rate of PET hydrolysis by SM14est was determined to be 0.004 s(−1) at 45°C, which was increased by 5-fold to 0.02 s(−1) upon addition of 500 mM sodium chloride. Sequence alignment and structural comparison with known PET hydrolases, including the marine halophile PET6, and the highly efficient, thermophilic PHL7, revealed conserved features of interest. Based on this work, SM14est emerges as a useful enzyme that is more similar to key players in the area of PET hydrolysis, like PHL7 and IsPETase, than it is to its marine counterparts. Salt-tolerant polyesterases such as SM14est are potentially valuable in the biological degradation of plastic particles that readily contaminate marine ecosystems and industrial wastewaters. Frontiers Media S.A. 2023-05-12 /pmc/articles/PMC10213408/ /pubmed/37250061 http://dx.doi.org/10.3389/fmicb.2023.1170880 Text en Copyright © 2023 Carr, Keller, Paul, Schubert, Clausen, Jensen, Clarke, Westh and Dobson. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Carr, Clodagh M.
Keller, Malene B.
Paul, Bijoya
Schubert, Sune W.
Clausen, Kristine S. R.
Jensen, Kenneth
Clarke, David J.
Westh, Peter
Dobson, Alan D. W.
Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14
title Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14
title_full Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14
title_fullStr Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14
title_full_unstemmed Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14
title_short Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14
title_sort purification and biochemical characterization of sm14est, a pet-hydrolyzing enzyme from the marine sponge-derived streptomyces sp. sm14
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10213408/
https://www.ncbi.nlm.nih.gov/pubmed/37250061
http://dx.doi.org/10.3389/fmicb.2023.1170880
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