β-sheet Engineering of IsPETase for PET Depolymerization Article Swipe
Songfeng Gao
,
Lixia Shi
,
Hongli Wei
,
Pi Liu
,
Wei Zhao
,
Ling Gong
,
Zijian Tan
,
Huanhuan Zhai
,
Weidong Liu
,
Haifeng Liu
,
Leilei Zhu
·
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.1016/j.eng.2024.10.015
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.1016/j.eng.2024.10.015
The enzymatic depolymerization of polyethylene terephthalate (PET) offers a sustainable approach for the recycling of PET waste. Great efforts have been devoted to engineering PET depolymerases on the substrate binding cleft and the surrounding loops/α-helices on the surface. Here, we report the systematic engineering of whole β-sheet regions in the core of IsPETase (a PETase from Ideonella sakaiensis) via a fluorescent high-throughput screening assay. Twenty-one beneficial substitutions were obtained and iteratively recombined. The best variant, DepoPETase β, with an increase in the melting temperatures (Tm) of 22.9 °C, exhibited superior depolymerization performance and enabled complete depolymerization of 100.5 g of untreated post-consumer PET (pc-PET; 0.26% Wenzyme/WPET enzyme loading) in liter-scale bioreactor at 50 °C within 4 d. Crystallization and molecular dynamics simulations revealed that the improved activity and thermostability of DepoPETase β were due to enhanced hydrogen bonds and salt bridges in the β-sheet region, a more tightly packed structure of the core sheets and the surrounding helix, and improved binding of PET to the active sites. This study not only demonstrates the importance of engineering strategy in the β-sheet region of PET hydrolases but also provides a potential PET depolymerase for large-scale PET recycling.
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Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1016/j.eng.2024.10.015
- OA Status
- gold
- Cited By
- 3
- References
- 101
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4404450274
All OpenAlex metadata
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https://openalex.org/W4404450274Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1016/j.eng.2024.10.015Digital Object Identifier
- Title
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β-sheet Engineering of IsPETase for PET DepolymerizationWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2024Year of publication
- Publication date
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2024-11-17Full publication date if available
- Authors
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Songfeng Gao, Lixia Shi, Hongli Wei, Pi Liu, Wei Zhao, Ling Gong, Zijian Tan, Huanhuan Zhai, Weidong Liu, Haifeng Liu, Leilei ZhuList of authors in order
- Landing page
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https://doi.org/10.1016/j.eng.2024.10.015Publisher landing page
- Open access
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YesWhether a free full text is available
- OA status
-
goldOpen access status per OpenAlex
- OA URL
-
https://doi.org/10.1016/j.eng.2024.10.015Direct OA link when available
- Concepts
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Depolymerization, Polymer science, Engineering, Materials science, Polymer chemistryTop concepts (fields/topics) attached by OpenAlex
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3Total citation count in OpenAlex
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2025: 3Per-year citation counts (last 5 years)
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101Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.(PET) | 6 |
| abstract_inverted_index.0.26% | 104 |
| abstract_inverted_index.100.5 | 97 |
| abstract_inverted_index.Great | 17 |
| abstract_inverted_index.Here, | 38 |
| abstract_inverted_index.bonds | 137 |
| abstract_inverted_index.cleft | 30 |
| abstract_inverted_index.study | 168 |
| abstract_inverted_index.whole | 45 |
| abstract_inverted_index.PETase | 54 |
| abstract_inverted_index.active | 165 |
| abstract_inverted_index.assay. | 63 |
| abstract_inverted_index.enzyme | 106 |
| abstract_inverted_index.helix, | 157 |
| abstract_inverted_index.offers | 7 |
| abstract_inverted_index.packed | 148 |
| abstract_inverted_index.region | 180 |
| abstract_inverted_index.report | 40 |
| abstract_inverted_index.sheets | 153 |
| abstract_inverted_index.sites. | 166 |
| abstract_inverted_index.waste. | 16 |
| abstract_inverted_index.within | 114 |
| abstract_inverted_index.binding | 29, 160 |
| abstract_inverted_index.bridges | 140 |
| abstract_inverted_index.devoted | 21 |
| abstract_inverted_index.efforts | 18 |
| abstract_inverted_index.enabled | 93 |
| abstract_inverted_index.melting | 82 |
| abstract_inverted_index.region, | 144 |
| abstract_inverted_index.regions | 47 |
| abstract_inverted_index.tightly | 147 |
| abstract_inverted_index.(pc-PET; | 103 |
| abstract_inverted_index.IsPETase | 52 |
| abstract_inverted_index.activity | 126 |
| abstract_inverted_index.approach | 10 |
| abstract_inverted_index.complete | 94 |
| abstract_inverted_index.dynamics | 120 |
| abstract_inverted_index.enhanced | 135 |
| abstract_inverted_index.hydrogen | 136 |
| abstract_inverted_index.improved | 125, 159 |
| abstract_inverted_index.increase | 79 |
| abstract_inverted_index.loading) | 107 |
| abstract_inverted_index.obtained | 68 |
| abstract_inverted_index.provides | 186 |
| abstract_inverted_index.revealed | 122 |
| abstract_inverted_index.strategy | 176 |
| abstract_inverted_index.superior | 89 |
| abstract_inverted_index.surface. | 37 |
| abstract_inverted_index.variant, | 74 |
| abstract_inverted_index.β-sheet | 46, 143, 179 |
| abstract_inverted_index.Ideonella | 56 |
| abstract_inverted_index.enzymatic | 1 |
| abstract_inverted_index.exhibited | 88 |
| abstract_inverted_index.molecular | 119 |
| abstract_inverted_index.potential | 188 |
| abstract_inverted_index.recycling | 13 |
| abstract_inverted_index.screening | 62 |
| abstract_inverted_index.structure | 149 |
| abstract_inverted_index.substrate | 28 |
| abstract_inverted_index.untreated | 100 |
| abstract_inverted_index.DepoPETase | 75, 130 |
| abstract_inverted_index.Twenty-one | 64 |
| abstract_inverted_index.beneficial | 65 |
| abstract_inverted_index.bioreactor | 110 |
| abstract_inverted_index.hydrolases | 183 |
| abstract_inverted_index.importance | 173 |
| abstract_inverted_index.recycling. | 194 |
| abstract_inverted_index.systematic | 42 |
| abstract_inverted_index.engineering | 23, 43, 175 |
| abstract_inverted_index.fluorescent | 60 |
| abstract_inverted_index.iteratively | 70 |
| abstract_inverted_index.large-scale | 192 |
| abstract_inverted_index.liter-scale | 109 |
| abstract_inverted_index.performance | 91 |
| abstract_inverted_index.recombined. | 71 |
| abstract_inverted_index.sakaiensis) | 57 |
| abstract_inverted_index.simulations | 121 |
| abstract_inverted_index.surrounding | 33, 156 |
| abstract_inverted_index.sustainable | 9 |
| abstract_inverted_index.Wenzyme/WPET | 105 |
| abstract_inverted_index.demonstrates | 171 |
| abstract_inverted_index.depolymerase | 190 |
| abstract_inverted_index.polyethylene | 4 |
| abstract_inverted_index.temperatures | 83 |
| abstract_inverted_index.depolymerases | 25 |
| abstract_inverted_index.post-consumer | 101 |
| abstract_inverted_index.substitutions | 66 |
| abstract_inverted_index.terephthalate | 5 |
| abstract_inverted_index.Crystallization | 117 |
| abstract_inverted_index.high-throughput | 61 |
| abstract_inverted_index.thermostability | 128 |
| abstract_inverted_index.depolymerization | 2, 90, 95 |
| abstract_inverted_index.loops/α-helices | 34 |
| cited_by_percentile_year.max | 97 |
| cited_by_percentile_year.min | 96 |
| countries_distinct_count | 0 |
| institutions_distinct_count | 11 |
| citation_normalized_percentile.value | 0.61181212 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | False |