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...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-05-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1170880/full |
| _version_ | 1797828909476085760 |
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| author | Clodagh M. Carr Clodagh M. Carr Malene B. Keller Bijoya Paul Sune W. Schubert Kristine S. R. Clausen Kenneth Jensen David J. Clarke David J. Clarke Peter Westh Alan D. W. Dobson Alan D. W. Dobson Alan D. W. Dobson |
| author_facet | Clodagh M. Carr Clodagh M. Carr Malene B. Keller Bijoya Paul Sune W. Schubert Kristine S. R. Clausen Kenneth Jensen David J. Clarke David J. Clarke Peter Westh Alan D. W. Dobson Alan D. W. Dobson Alan D. W. Dobson |
| author_sort | Clodagh M. Carr |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-09T13:11:57Z |
| format | Article |
| id | doaj.art-087d005c997a4245819954d6b162183e |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-09T13:11:57Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-087d005c997a4245819954d6b162183e2023-05-12T06:06:30ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2023-05-011410.3389/fmicb.2023.11708801170880Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14Clodagh M. Carr0Clodagh M. Carr1Malene B. Keller2Bijoya Paul3Sune W. Schubert4Kristine S. R. Clausen5Kenneth Jensen6David J. Clarke7David J. Clarke8Peter Westh9Alan D. W. Dobson10Alan D. W. Dobson11Alan D. W. Dobson12School of Microbiology, University College Cork, Cork, IrelandSSPC-SFI Research Centre for Pharmaceuticals, University College Cork, Cork, IrelandDepartment of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, DenmarkDepartment of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, DenmarkDepartment of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, DenmarkDepartment of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, DenmarkNovozymes A/S, Lyngby, DenmarkSchool of Microbiology, University College Cork, Cork, IrelandAPC Microbiome Ireland, University College Cork, Cork, IrelandDepartment of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, DenmarkSchool of Microbiology, University College Cork, Cork, IrelandSSPC-SFI Research Centre for Pharmaceuticals, University College Cork, Cork, IrelandEnvironmental Research Institute, University College Cork, Cork, IrelandThe 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.https://www.frontiersin.org/articles/10.3389/fmicb.2023.1170880/fullplasticmarinePETasebiorecyclingbioremediation |
| spellingShingle | Clodagh M. Carr Clodagh M. Carr Malene B. Keller Bijoya Paul Sune W. Schubert Kristine S. R. Clausen Kenneth Jensen David J. Clarke David J. Clarke Peter Westh Alan D. W. Dobson Alan D. W. Dobson Alan D. W. Dobson Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14 Frontiers in Microbiology plastic marine PETase biorecycling bioremediation |
| 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 | plastic marine PETase biorecycling bioremediation |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1170880/full |
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