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Here we computationally incorporated two hydrolytic active sites with geometries similar to that of Idionella sakaiensis PET hydrolase, to fragaceatoxin C (FraC), a membrane pore-forming protein. FraCm1/m2 could be assembled into octameric nanopores (7.0 nm high × 1.6–6.0 nm entry), which deconstructed (40 °C, pH 7.0) nPET from GoodFellow, commodities and plastic bottles. FraCm1 and FraCm2 degrade nPET by endo- and exo-type chain scission. While FraCm1 produces bis(2-hydroxyethyl) terephthalate as the main product, FraCm2 yields a high diversity of oligomers and terephthalic acid. Mechanistic and biochemical differences with benchmark PET hydrolases, along with pore and nPET dynamics, suggest that these pore-forming protein catalytic nanoreactors do not deconstruct macro-PET but are promising in nanotechnology for filtering, capturing and breaking down nPET, for example, in wastewater treatment plants. \n \n \n \n The development of innovative strategies for the capture and biodegradation of nanoplastics is sought after. 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data-test="article-category">Article</li> <li class="c-article-identifiers__item"> <a href="https://www.springernature.com/gp/open-research/about/the-fundamentals-of-open-access-and-open-research" data-track="click" data-track-action="open access" data-track-label="link" class="u-color-open-access" data-test="open-access">Open access</a> </li> <li class="c-article-identifiers__item">Published: <time datetime="2023-10-19">19 October 2023</time></li> </ul> <h1 class="c-article-title" data-test="article-title" data-article-title="">Sub-micro- and nano-sized polyethylene terephthalate deconstruction with engineered protein nanopores</h1> <ul class="c-article-author-list c-article-author-list--short" data-test="authors-list" data-component-authors-activator="authors-list"><li class="c-article-author-list__item"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-Ana-Robles_Mart_n-Aff1-Aff2" data-author-popup="auth-Ana-Robles_Mart_n-Aff1-Aff2" data-author-search="Robles-Martín, Ana">Ana Robles-Martín</a><span class="u-js-hide"> <a class="js-orcid" href="http://orcid.org/0000-0002-6377-6338"><span class="u-visually-hidden">ORCID: </span>orcid.org/0000-0002-6377-6338</a></span><sup class="u-js-hide"><a href="#Aff1">1</a>,<a href="#Aff2">2</a></sup><sup class="u-js-hide"> <a href="#na1">na1</a></sup>, </li><li class="c-article-author-list__item"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-Rafael-Amigot_S_nchez-Aff3" data-author-popup="auth-Rafael-Amigot_S_nchez-Aff3" data-author-search="Amigot-Sánchez, Rafael">Rafael Amigot-Sánchez</a><span class="u-js-hide"> <a class="js-orcid" href="http://orcid.org/0000-0001-9253-1631"><span class="u-visually-hidden">ORCID: </span>orcid.org/0000-0001-9253-1631</a></span><sup class="u-js-hide"><a href="#Aff3">3</a></sup><sup class="u-js-hide"> <a 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class="u-visually-hidden">pages </span>1174–1185 (<span data-test="article-publication-year">2023</span>)<a href="#citeas" class="c-article-info-details__cite-as u-hide-print" data-track="click" data-track-action="cite this article" data-track-label="link">Cite this article</a> </p> <div class="c-article-metrics-bar__wrapper u-clear-both"> <ul class="c-article-metrics-bar u-list-reset"> <li class=" c-article-metrics-bar__item" data-test="access-count"> <p class="c-article-metrics-bar__count">14k <span class="c-article-metrics-bar__label">Accesses</span></p> </li> <li class="c-article-metrics-bar__item" data-test="citation-count"> <p class="c-article-metrics-bar__count">6 <span class="c-article-metrics-bar__label">Citations</span></p> </li> <li class="c-article-metrics-bar__item" data-test="altmetric-score"> <p class="c-article-metrics-bar__count">190 <span class="c-article-metrics-bar__label">Altmetric</span></p> </li> <li class="c-article-metrics-bar__item"> <p class="c-article-metrics-bar__details"><a href="/articles/s41929-023-01048-6/metrics" data-track="click" data-track-action="view metrics" data-track-label="link" rel="nofollow">Metrics <span class="u-visually-hidden">details</span></a></p> </li> </ul> </div> </header> <div class="u-js-hide" data-component="article-subject-links"> <h3 class="c-article__sub-heading">Subjects</h3> <ul class="c-article-subject-list"> <li class="c-article-subject-list__subject"><a href="/subjects/nanopores" data-track="click" data-track-action="view subject" data-track-label="link">Nanopores</a></li><li class="c-article-subject-list__subject"><a href="/subjects/protein-design" data-track="click" data-track-action="view subject" data-track-label="link">Protein design</a></li> </ul> </div> </div> <div class="c-article-body"> <section aria-labelledby="Abs1" data-title="Abstract" lang="en"><div class="c-article-section" id="Abs1-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Abs1">Abstract</h2><div class="c-article-section__content" id="Abs1-content"><p>The identification or design of biocatalysts to mitigate the accumulation of plastics, including sub-micro- and nano-sized polyethylene terephthalate (nPET), is becoming a global challenge. Here we computationally incorporated two hydrolytic active sites with geometries similar to that of <i>Idionella sakaiensis</i> PET hydrolase, to fragaceatoxin C (FraC), a membrane pore-forming protein. FraC<sub>m1/m2</sub> could be assembled into octameric nanopores (7.0 nm high × 1.6–6.0 nm entry), which deconstructed (40 °C, pH 7.0) nPET from GoodFellow, commodities and plastic bottles. FraC<sub>m1</sub> and FraC<sub>m2</sub> degrade nPET by endo- and exo-type chain scission. While FraC<sub>m1</sub> produces bis(2-hydroxyethyl) terephthalate as the main product, FraC<sub>m2</sub> yields a high diversity of oligomers and terephthalic acid. Mechanistic and biochemical differences with benchmark PET hydrolases, along with pore and nPET dynamics, suggest that these pore-forming protein catalytic nanoreactors do not deconstruct macro-PET but are promising in nanotechnology for filtering, capturing and breaking down nPET, for example, in wastewater treatment plants.</p><div class="c-article-section__figure" data-test="figure" data-container-section="figure"><figure><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Figa_HTML.png?as=webp"><img src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Figa_HTML.png" alt="" loading="lazy" width="685" height="417"></picture></div></div></figure></div></div></div></section> <noscript> </noscript> <section aria-labelledby="inline-recommendations" data-title="Inline Recommendations" class="c-article-recommendations" data-track-component="inline-recommendations"> <h3 class="c-article-recommendations-title" id="inline-recommendations">Similar content being viewed by others</h3> <div class="c-article-recommendations-list"> <div class="c-article-recommendations-list__item"> <article class="c-article-recommendations-card" itemscope itemtype="http://schema.org/ScholarlyArticle"> <div class="c-article-recommendations-card__img"><img src="https://media.springernature.com/w215h120/springer-static/image/art%3A10.1038%2Fs41467-024-47007-y/MediaObjects/41467_2024_47007_Fig1_HTML.png" loading="lazy" alt=""></div> <div class="c-article-recommendations-card__main"> <h3 class="c-article-recommendations-card__heading" itemprop="name headline"> <a class="c-article-recommendations-card__link" itemprop="url" href="https://www.nature.com/articles/s41467-024-47007-y?fromPaywallRec=false" data-track="select_recommendations_1" data-track-context="inline recommendations" data-track-action="click recommendations inline - 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Adapting to plastic.Nat. Rev. Microbiol. 18, 362–363 (2020)." href="/articles/s41929-023-01048-6#ref-CR1" id="ref-link-section-d10881139e669">1</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 2" title="Rosenboom, J. G., Langer, R. & Traverso, G. Bioplastics for a circular economy. Nat. Rev. Mater. 7, 117–137 (2022)." href="/articles/s41929-023-01048-6#ref-CR2" id="ref-link-section-d10881139e672">2</a></sup>, making them the most common waste material in the world, accounting for >380 million tons. Their cumulative greenhouse gas emissions from production to disposal may be equivalent to 10–13% of the total carbon budget; 10.2% are predicted to correspond to polyethylene terephthalate (PET) plastics<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 3" title="Geyer, R., Jambeck, J. R. & Law, K. L. Production, use, and fate of all plastics ever made. Sci. Adv. 3, e1700782 (2017)." href="/articles/s41929-023-01048-6#ref-CR3" id="ref-link-section-d10881139e676">3</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 4" title="Cabernard, L. et al. Growing environmental footprint of plastics driven by coal combustion. Nat. Sustain. 5, 139–148 (2022)." href="/articles/s41929-023-01048-6#ref-CR4" id="ref-link-section-d10881139e679">4</a></sup>. Approximately 80% of waste objects are composed of macroplastics, with PET accounting for 14.4% of total plastic waste<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="York, A. Adapting to plastic.Nat. Rev. Microbiol. 18, 362–363 (2020)." href="#ref-CR1" id="ref-link-section-d10881139e683">1</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Rosenboom, J. G., Langer, R. & Traverso, G. Bioplastics for a circular economy. Nat. Rev. Mater. 7, 117–137 (2022)." href="#ref-CR2" id="ref-link-section-d10881139e683_1">2</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Geyer, R., Jambeck, J. R. & Law, K. L. Production, use, and fate of all plastics ever made. Sci. Adv. 3, e1700782 (2017)." href="#ref-CR3" id="ref-link-section-d10881139e683_2">3</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 4" title="Cabernard, L. et al. Growing environmental footprint of plastics driven by coal combustion. Nat. Sustain. 5, 139–148 (2022)." href="/articles/s41929-023-01048-6#ref-CR4" id="ref-link-section-d10881139e686">4</a></sup>, which can reach the oceans from land<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 2" title="Rosenboom, J. G., Langer, R. & Traverso, G. Bioplastics for a circular economy. Nat. Rev. Mater. 7, 117–137 (2022)." href="/articles/s41929-023-01048-6#ref-CR2" id="ref-link-section-d10881139e690">2</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 5" title="Jambeck, J. R. et al. Marine pollution. Plastic waste inputs from land into the ocean. Science 347, 768–771 (2015)." href="/articles/s41929-023-01048-6#ref-CR5" id="ref-link-section-d10881139e693">5</a></sup>, and be degraded over time to microplastics (5 mm to 1 µm), sub-microplastics (100 nm to 1 µm), and nanoplastics (1 nm to 100 nm)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 1" title="York, A. Adapting to plastic.Nat. Rev. Microbiol. 18, 362–363 (2020)." href="/articles/s41929-023-01048-6#ref-CR1" id="ref-link-section-d10881139e697">1</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 2" title="Rosenboom, J. G., Langer, R. & Traverso, G. Bioplastics for a circular economy. Nat. Rev. Mater. 7, 117–137 (2022)." href="/articles/s41929-023-01048-6#ref-CR2" id="ref-link-section-d10881139e700">2</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 6" title="Gigault, J. et al. Current opinion: what is a nanoplastic? Environ. Pollut. 235, 1030–1034 (2018)." href="/articles/s41929-023-01048-6#ref-CR6" id="ref-link-section-d10881139e703">6</a></sup>. Recent studies confirmed that PET particles comprise at least 5% of the total identified plastic particles<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 1" title="York, A. Adapting to plastic.Nat. Rev. Microbiol. 18, 362–363 (2020)." href="/articles/s41929-023-01048-6#ref-CR1" id="ref-link-section-d10881139e708">1</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 2" title="Rosenboom, J. G., Langer, R. & Traverso, G. Bioplastics for a circular economy. Nat. Rev. Mater. 7, 117–137 (2022)." href="/articles/s41929-023-01048-6#ref-CR2" id="ref-link-section-d10881139e711">2</a></sup>, which can be found in concentrations equivalent to trillions of particles in the air of some cities<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 7" title="Allen, S. et al. Atmospheric transport and deposition of microplastics in a remote mountain catchment. Nat. Geosci. 12, 339–344 (2019)." href="/articles/s41929-023-01048-6#ref-CR7" id="ref-link-section-d10881139e715">7</a></sup>, from 2,649 to 6,292 particles per litre in mineral water<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 8" title="Schymanski, D. et al. Analysis of microplastics in drinking water and other clean water samples with micro-Raman and micro-infrared spectroscopy: minimum requirements and best practice guidelines. Anal. Bioanal. Chem. 413, 5969–5994 (2021)." href="/articles/s41929-023-01048-6#ref-CR8" id="ref-link-section-d10881139e719">8</a></sup>, and from 5 to 52.3 ng ml<sup>−1</sup> in mountain and Antarctica ice samples<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 9" title="Materić, D. et al. Nanoplastics measurements in Northern and Southern polar ice. Environ. Res. 208, 112741 (2022)." href="/articles/s41929-023-01048-6#ref-CR9" id="ref-link-section-d10881139e725">9</a></sup>. The ubiquity of plastic debris is causing an unprecedented ecological crisis<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 1" title="York, A. Adapting to plastic.Nat. Rev. Microbiol. 18, 362–363 (2020)." href="/articles/s41929-023-01048-6#ref-CR1" id="ref-link-section-d10881139e729">1</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 2" title="Rosenboom, J. G., Langer, R. & Traverso, G. Bioplastics for a circular economy. Nat. Rev. Mater. 7, 117–137 (2022)." href="/articles/s41929-023-01048-6#ref-CR2" id="ref-link-section-d10881139e732">2</a></sup>.</p><p>PET recycling is theoretically attainable<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 10" title="Wei, R., Bertling, J., O’Connor, K., Bank, L. M. & Bornscheur, U. T. Possibilities and limitations of biotechnological plastic degradation and recycling. Nat. Catal. 3, 867–871 (2020)." href="/articles/s41929-023-01048-6#ref-CR10" id="ref-link-section-d10881139e739">10</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 11" title="Ellis, L. D. et al. Chemical and biological catalysis for plastics recycling and upcycling. Nat. Catal. 4, 539–556 (2021)." href="/articles/s41929-023-01048-6#ref-CR11" id="ref-link-section-d10881139e742">11</a></sup>, compared with other extremely challenging plastics, such as thermosets<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 6" title="Gigault, J. et al. Current opinion: what is a nanoplastic? Environ. Pollut. 235, 1030–1034 (2018)." href="/articles/s41929-023-01048-6#ref-CR6" id="ref-link-section-d10881139e746">6</a></sup>. While there are more widely used and better established chemical recycling methods for PET<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 10" title="Wei, R., Bertling, J., O’Connor, K., Bank, L. M. & Bornscheur, U. T. Possibilities and limitations of biotechnological plastic degradation and recycling. Nat. Catal. 3, 867–871 (2020)." href="/articles/s41929-023-01048-6#ref-CR10" id="ref-link-section-d10881139e750">10</a></sup>, enzymatic PET recycling and upcycling strategies have been demonstrated as necessary alternatives<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 10" title="Wei, R., Bertling, J., O’Connor, K., Bank, L. M. & Bornscheur, U. T. Possibilities and limitations of biotechnological plastic degradation and recycling. Nat. Catal. 3, 867–871 (2020)." href="/articles/s41929-023-01048-6#ref-CR10" id="ref-link-section-d10881139e754">10</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 11" title="Ellis, L. D. et al. Chemical and biological catalysis for plastics recycling and upcycling. Nat. Catal. 4, 539–556 (2021)." href="/articles/s41929-023-01048-6#ref-CR11" id="ref-link-section-d10881139e757">11</a></sup>. Over the past 18 years, only a few benchmark enzymes from cultured and uncultured microorganisms have been shown to degrade macro-PET<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 12" title="Wei, R. et al. Mechanism-based design of efficient PET hydrolases. ACS Catal. 12, 3382–3396 (2022)." href="/articles/s41929-023-01048-6#ref-CR12" id="ref-link-section-d10881139e761">12</a></sup>, some of which have been engineered to increase the stability (up to +37.5 °C) and activity at temperatures close to or exceeding the PET glass transition temperature (<i>T</i><sub>G</sub>), <span class="stix">∼</span>70 °C. This temperature is recommended to access amorphous regions of PET polymer<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 12" title="Wei, R. et al. Mechanism-based design of efficient PET hydrolases. ACS Catal. 12, 3382–3396 (2022)." href="/articles/s41929-023-01048-6#ref-CR12" id="ref-link-section-d10881139e770">12</a></sup>, as it is physically impossible to enzymatically degrade crystalline PET, having a melting temperature (<i>T</i><sub>m</sub>) of 260 °C (Supplementary Note <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">1</a>). Notable engineered examples are ThermoPETase<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 13" title="Son, H. F. et al. Rational protein engineering of thermo-stable PETase from Ideonella sakaiensis for highly efficient PET degradation. ACS Catal. 9, 3519–3526 (2019)." href="/articles/s41929-023-01048-6#ref-CR13" id="ref-link-section-d10881139e781">13</a></sup> and HotPETase<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 14" title="Bell, E. L. et al. Directed evolution of an efficient and thermostable PET depolymerase. Nat. Catal. 5, 673–681 (2022)." href="/articles/s41929-023-01048-6#ref-CR14" id="ref-link-section-d10881139e786">14</a></sup> from <i>Idionella sakaiensis</i>, and LCC<sub>ICCG</sub> and LCC<sub>WCCG</sub> from leaf-branch compost<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 15" title="Tournier, V. et al. An engineered PET depolymerase to break down and recycle plastic bottles. Nature 580, 216–219 (2020)." href="/articles/s41929-023-01048-6#ref-CR15" id="ref-link-section-d10881139e797">15</a></sup>. Although challenging<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 12" title="Wei, R. et al. Mechanism-based design of efficient PET hydrolases. ACS Catal. 12, 3382–3396 (2022)." href="/articles/s41929-023-01048-6#ref-CR12" id="ref-link-section-d10881139e801">12</a></sup>, the degradation of PET below <i>T</i><sub>G</sub> has also been reported, notably, for the machine learning-engineered variant FAST-PETase that completely depolymerizes at 50 °C untreated postconsumer-PET<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 16" title="Lu, H. et al. Machine learning-aided engineering of hydrolases for PET depolymerization. Nature 604, 662–667 (2022)." href="/articles/s41929-023-01048-6#ref-CR16" id="ref-link-section-d10881139e810">16</a></sup>, and for the <i>I. sakaiensis</i> PETase (<i>Is</i>PETase), active at 30 °C but labile at 37 °C after 24 h of incubation<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 12" title="Wei, R. et al. Mechanism-based design of efficient PET hydrolases. ACS Catal. 12, 3382–3396 (2022)." href="/articles/s41929-023-01048-6#ref-CR12" id="ref-link-section-d10881139e820">12</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 17" title="Yoshida, S. et al. A bacterium that degrades and assimilates poly(ethylene terephthalate). Science 351, 1196–1199 (2016)." href="/articles/s41929-023-01048-6#ref-CR17" id="ref-link-section-d10881139e823">17</a></sup>, and its redesigned variant DuraPETase that is long<i>-</i>term active at 37 °C (ref. <sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 18" title="Cui, Y. et al. Computational redesign of a PETase for plastic biodegradation under ambient condition by the GRAPE strategy. ACS Catal. 11, 1340–1350 (2021)." href="/articles/s41929-023-01048-6#ref-CR18" id="ref-link-section-d10881139e831">18</a></sup>).</p><p>Micro-, sub-micro- and nano-PET, hereinafter referred to as nPET, pollution has also been targeted for enzymatic deconstruction. DuraPETase degrades PET microplastics (10–50 µm) and nanoplastics (50–100 nm) at concentrations (0.2–0.3 g l<sup>−1</sup>) 100-fold higher than those in wastewater plants, within 20 days at 37 °C (micro-) and within 1 h at 37 °C (nano-)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 18" title="Cui, Y. et al. Computational redesign of a PETase for plastic biodegradation under ambient condition by the GRAPE strategy. ACS Catal. 11, 1340–1350 (2021)." href="/articles/s41929-023-01048-6#ref-CR18" id="ref-link-section-d10881139e840">18</a></sup>. The PETase from <i>Thermobifida fusca</i>, <i>Tf</i>Cut2, degrades nPET (100–164 nm) at 60 °C (refs. <sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 19" title="Barth, M. et al. Effect of hydrolysis products on the enzymatic degradation of polyethylene terephthalate nanoparticles by a polyester hydrolase from Thermobifida fusca. Biochem. Eng. J. 93, 222–228 (2015)." href="/articles/s41929-023-01048-6#ref-CR19" id="ref-link-section-d10881139e850">19</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 20" title="Vogel, K. et al. Enzymatic degradation of polyethylene terephthalate nanoplastics analyzed in real time by isothermal titration calorimetry. Sci. Total Environ. 773, 145111 (2021)." href="/articles/s41929-023-01048-6#ref-CR20" id="ref-link-section-d10881139e853">20</a></sup>). These studies demonstrated the potential of PETases to also ameliorate nPET pollution, a line that we want to explore in this study, by designing a protein capable of filtering and degrading nPET. Within this context, actinoporins constitute a group of small and basic α pore-forming toxins produced by sea anemones as a defence mechanism<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 21" title="Tanaka, K., Caaveiro, J. M. M., Morante, K., González-Mañas, J. M. & Tsumoto, K. Structural basis for self-assembly of a cytolytic pore lined by protein and lipid. Nat. Commun. 6, 1–11 (2015)." href="/articles/s41929-023-01048-6#ref-CR21" id="ref-link-section-d10881139e858">21</a></sup>. In water, these easily produced and purified non-catalytic proteins remain perfectly soluble and stably folded; however, upon interaction with lipid membranes of a specific composition, they spontaneously become oligomeric integral membrane structures and make a pore<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 22" title="Alegre-Cebollada, J. et al. Silent mutations at the 5′-end of the cDNA of actinoporins from the sea anemone Stichodactyla helianthus allow their heterologous overproduction in Escherichia coli. J. Biotechnol. 127, 211–221 (2007)." href="/articles/s41929-023-01048-6#ref-CR22" id="ref-link-section-d10881139e862">22</a></sup>. Because of their physicochemical properties and configuration, most efforts towards designing pore-forming toxins have been directed at sensing molecules and sequencing<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 23" title="Wloka, C., Mutter, N. L., Soskine, M. & Maglia, G. Alpha-helical Fragaceatoxin C nanopore engineered for double-stranded and single-stranded nucleic acid analysis. Angew. Chem. Int. Ed. Engl. 55, 12494–12498 (2016)." href="/articles/s41929-023-01048-6#ref-CR23" id="ref-link-section-d10881139e866">23</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 24" title="Versloot, R. C. A. et al. Quantification of protein glycosylation using nanopores. Nano Lett. 22, 5357–5364 (2022)." href="/articles/s41929-023-01048-6#ref-CR24" id="ref-link-section-d10881139e869">24</a></sup>.</p><p>In this Article, we generated a pore-forming catalytic enzyme that could be further assembled into catalytic nanopores/nanoreactors that act as a reaction chamber to degrade nPET. This advance can be addressed due to recent modelling developments in designing de novo active sites<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 25" title="Roda, S., Robles-Martín, A., Xiang, R., Kazemi, M. & Guallar, V. Structural-based modeling in protein engineering. A must do. J. Phys. Chem. B 125, 6491–6500 (2021)." href="/articles/s41929-023-01048-6#ref-CR25" id="ref-link-section-d10881139e876">25</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 26" title="Alonso, S. et al. Genetically engineered proteins with two active sites for enhanced biocatalysis and synergistic chemo- and biocatalysis. Nat. Catal. 3, 319–328 (2020)." href="/articles/s41929-023-01048-6#ref-CR26" id="ref-link-section-d10881139e879">26</a></sup>. As a target, we selected fragaceatoxin C (FraC) from <i>Actinia fragacea</i>, the only actinoporin membrane pore with a three-dimensional structure solved with atomic resolution<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 21" title="Tanaka, K., Caaveiro, J. M. M., Morante, K., González-Mañas, J. M. & Tsumoto, K. Structural basis for self-assembly of a cytolytic pore lined by protein and lipid. Nat. Commun. 6, 1–11 (2015)." href="/articles/s41929-023-01048-6#ref-CR21" id="ref-link-section-d10881139e886">21</a></sup>. FraC is a stable 7-nm-high octameric V-shaped pore formed by spontaneous self-assembly of the protein monomers and with access sizes on the order of 6.7 nm in the <i>cis</i> entry and on the order of 1.9 nm in the <i>trans</i> exit. A computational structure-based modelling method, the <i>PluriZyme</i> strategy<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 26" title="Alonso, S. et al. Genetically engineered proteins with two active sites for enhanced biocatalysis and synergistic chemo- and biocatalysis. Nat. Catal. 3, 319–328 (2020)." href="/articles/s41929-023-01048-6#ref-CR26" id="ref-link-section-d10881139e900">26</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 27" title="Roda, S. et al. A Plurizyme with transaminase and hydrolase activity catalyzes cascade reactions. Angew. Chem. Int. Ed. Engl. 61, e202207344 (2022)." href="/articles/s41929-023-01048-6#ref-CR27" id="ref-link-section-d10881139e903">27</a></sup>, was applied to add an artificial catalytic serine–histidine–aspartic triad and an oxyanion hole capable of ester hydrolysis to the non-catalytic FraC. These catalytic elements match the active sites of PETases<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 12" title="Wei, R. et al. Mechanism-based design of efficient PET hydrolases. ACS Catal. 12, 3382–3396 (2022)." href="/articles/s41929-023-01048-6#ref-CR12" id="ref-link-section-d10881139e907">12</a></sup>. When two newly engineered catalytic proteins (FraC<sub>m1</sub> and FraC<sub>m2</sub>) were assembled as nanopores (npFraC<sub>m1</sub> and npFraC<sub>m2</sub>) they became pore-based nanoreactors for the depolymerization of nPET. We discovered different degradation product profiles between them and in comparison with benchmark PETases, namely LCC<sub>WT</sub>, LCC<sub>WCCG</sub> and <i>Is</i>PETase<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 12" title="Wei, R. et al. Mechanism-based design of efficient PET hydrolases. ACS Catal. 12, 3382–3396 (2022)." href="/articles/s41929-023-01048-6#ref-CR12" id="ref-link-section-d10881139e927">12</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 15" title="Tournier, V. et al. An engineered PET depolymerase to break down and recycle plastic bottles. Nature 580, 216–219 (2020)." href="/articles/s41929-023-01048-6#ref-CR15" id="ref-link-section-d10881139e930">15</a></sup>.</p></div></div></section><section data-title="Results"><div class="c-article-section" id="Sec2-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec2">Results</h2><div class="c-article-section__content" id="Sec2-content"><h3 class="c-article__sub-heading" id="Sec3">In silico design of catalytic pore-forming proteins</h3><p>The entire inner surface of the wild-type FraC pore structure, FraC<sub>WT</sub>, was explored using Protein Energy Landscape Exploration (PELE) software<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 26" title="Alonso, S. et al. Genetically engineered proteins with two active sites for enhanced biocatalysis and synergistic chemo- and biocatalysis. Nat. Catal. 3, 319–328 (2020)." href="/articles/s41929-023-01048-6#ref-CR26" id="ref-link-section-d10881139e948">26</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 27" title="Roda, S. et al. A Plurizyme with transaminase and hydrolase activity catalyzes cascade reactions. Angew. Chem. Int. Ed. Engl. 61, e202207344 (2022)." href="/articles/s41929-023-01048-6#ref-CR27" id="ref-link-section-d10881139e951">27</a></sup>, using as probes three esters (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">1</a>) commonly hydrolysed by most esterases and lipases<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 26" title="Alonso, S. et al. Genetically engineered proteins with two active sites for enhanced biocatalysis and synergistic chemo- and biocatalysis. Nat. Catal. 3, 319–328 (2020)." href="/articles/s41929-023-01048-6#ref-CR26" id="ref-link-section-d10881139e958">26</a></sup>. As in PluriZyme designs<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 26" title="Alonso, S. et al. Genetically engineered proteins with two active sites for enhanced biocatalysis and synergistic chemo- and biocatalysis. Nat. Catal. 3, 319–328 (2020)." href="/articles/s41929-023-01048-6#ref-CR26" id="ref-link-section-d10881139e962">26</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 27" title="Roda, S. et al. A Plurizyme with transaminase and hydrolase activity catalyzes cascade reactions. Angew. Chem. Int. Ed. Engl. 61, e202207344 (2022)." href="/articles/s41929-023-01048-6#ref-CR27" id="ref-link-section-d10881139e965">27</a></sup>, the initial goal is applying PELE to identify substrate binding sites for designing artificial hydrolase active sites. Energetic profiles of FraC<sub>WT</sub> from these simulations (Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">1</a>) are shown in Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig1">1a–c</a>, in which the binding energy minima are split into three regions based on the distance to the <i>trans</i> exit of the pore. Notably, the three substrates found good pockets at similar distances, denoting a suitable ester stabilizing environment. As shown in Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig1">1d</a>, from the <i>cis</i> side to the <i>trans</i> side, the outer cluster corresponds to the globular domain of the monomer at 70 Å from the bottom side (a more solvent-exposed side). The second cluster, located at a distance between 35 Å and 55 Å, corresponds to the hinge region of the monomer and is characterized by the presence of highly conserved residues involved in the conformational change forming the nanopore. We discovered several minima in this region but rejected them to avoid a loss of pore-forming activity<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 28" title="Mesa-Galloso et al. Disrupting a key hydrophobic pair in the oligomerization interface of the actinoporins impairs their pore-forming activity. Protein Sci. 26, 550–565 (2017)." href="/articles/s41929-023-01048-6#ref-CR28" id="ref-link-section-d10881139e991">28</a></sup>. At a distance of 20 Å, we discovered the transmembrane region configured by the N-terminal α-helices, where pockets are formed by non-conserved polar side chains facing the inner part of the channel. Both the first cluster and third cluster fulfilled our goal of introducing hydrolytic sites into the main pore, the dimensions/shapes of which can assist in channelling and holding substrates once they are assembled.</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-1" data-title="Global exploration of FraC binding site pockets using PELE."><figure><figcaption><b id="Fig1" class="c-article-section__figure-caption" data-test="figure-caption-text">Fig. 1: Global exploration of FraC binding site pockets using PELE.</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/1" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig1_HTML.png?as=webp"><img aria-describedby="Fig1" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig1_HTML.png" alt="figure 1" loading="lazy" width="685" height="626"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-1-desc"><p><b>a</b>, Energy profile of the global exploration with the branched ester glyceryl tripropionate and poses highlighted with the colour from each binding site (poses are highlighted only when the interaction energy is equal to or below −15 kcal mol<sup>−</sup><sup>1</sup>). <b>b</b>, Energy profile of the global exploration with the aromatic small-sized phenyl acetate and poses highlighted with the colour from each binding site (poses are only highlighted when the interaction energy is equal to or below −12.5 kcal mol<sup>−</sup><sup>1</sup>). <b>c</b>, Energy profile of the global exploration with the short chain alkenyl ester vinyl acetate and poses highlighted with the colour from each binding site (poses are highlighted only when the interaction energy is equal to or below −10 kcal mol<sup>−</sup><sup>1</sup>). <b>d</b>, Cross-sectional representation of FraC with two opposing chains to visualize the localization of the different indicated binding sites (Protein Data Bank (PDB) ID: 4TSY). Computational data were collected and analysed with PELE. Calculations and raw data are shown in Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">1</a>.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/1" data-track-dest="link:Figure1 Full size image" aria-label="Full size image figure 1" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><p>Interestingly, two acidic residues, Asp17 and Glu24, lie in the vicinity of the identified transmembrane pocket. In general, the insertion of a negative charge into a structure is detrimental, so we typically take advantage of the presence (if any) of acidic residues. Following this approach, the Lys20His and Thr21Ser variant introduced intra- and interhelix catalytic triad hydrogen bonds. Regarding the outer pocket, we also identified several acidic residues, Asp38, Glu40 and Glu173. In this case, there is also a native histidine, His175; PELE simulations revealed that the Asp38Ser variant could generate a catalytic triad. Moreover, we wanted to add a mutation that could act as an oxyanion hole to stabilize the negative charge that appears during hydrolysis: Glu173Gln.</p><p>The potential catalytic poses involving an ester carbon-serine oxygen distance <4.5 Å concurrent with hydrogen bonds between Ser–His and His–Asp/Glu were quantified for the three exploration model esters and the two variants, FraC<sub>m1</sub>, including Lys20His and Thr21Ser, and FraC<sub>m2</sub>, including Asp38Ser and Glu173Gln. The computed PELE-normalized relative catalytic activity ranged from 19.01% to 10.98% for FraC<sub>m1</sub> and from 2.05% to 1.42% for FraC<sub>m2</sub> (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig2">2a</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">1</a>). Furthermore, the computed PELE-normalized relative catalytic activity for bis(2-hydroxyethyl)-terephthalate (or <b>ETE</b>, following the nomenclature by Schubert et al.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 29" title="Schubert, S. et al. Reaction pathways for the enzymatic degradation of poly(ethylene terephthalate): What characterizes an efficient PET-hydrolase? ChemBioChem 24, e202200516 (2023)." href="/articles/s41929-023-01048-6#ref-CR29" id="ref-link-section-d10881139e1063">29</a></sup>), and mono-(2-hydroxyethyl)-terephthalic acid (or <b>TE</b>), which are incomplete degradation products during the hydrolysis of PET by PETases<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 12" title="Wei, R. et al. Mechanism-based design of efficient PET hydrolases. ACS Catal. 12, 3382–3396 (2022)." href="/articles/s41929-023-01048-6#ref-CR12" id="ref-link-section-d10881139e1070">12</a></sup> (Extended Data Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig8">1</a>), ranged from 8.79% to 3.61% for FraC<sub>m1</sub> and from 0.47% to 1.10% for FraC<sub>m2</sub> (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig2">2a</a>). FraC<sub>m1/m2</sub> active sites can thus potentially accommodate and convert ester substrates, including <b>TE</b> and/or <b>ETE</b>. Note that although both our designs (FraC<sub>m1/m2</sub>) and <i>Is</i>PETase<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 30" title="Jerves, C. et al. Reaction mechanism of the PET degrading enzyme PETase studied with DFT/MM molecular dynamics simulations. ACS Catal. 11, 11626–11638 (2021)." href="/articles/s41929-023-01048-6#ref-CR30" id="ref-link-section-d10881139e1099">30</a></sup> share an analogous active site geometry (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">2</a>), as seen by clarifying the enzyme–substrate encounter complex by quantum/molecular mechanics (QM/MM) (Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">1</a>), the overall FraC<sub>m1/m2</sub> pore is formed by eight different chains (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig2">2b</a>)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 21" title="Tanaka, K., Caaveiro, J. M. M., Morante, K., González-Mañas, J. M. & Tsumoto, K. Structural basis for self-assembly of a cytolytic pore lined by protein and lipid. Nat. Commun. 6, 1–11 (2015)." href="/articles/s41929-023-01048-6#ref-CR21" id="ref-link-section-d10881139e1115">21</a></sup>, thus introducing eight potential catalytic triads after pore assembly compared with the monomeric <i>Is</i>PETase<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 30" title="Jerves, C. et al. Reaction mechanism of the PET degrading enzyme PETase studied with DFT/MM molecular dynamics simulations. ACS Catal. 11, 11626–11638 (2021)." href="/articles/s41929-023-01048-6#ref-CR30" id="ref-link-section-d10881139e1122">30</a></sup>.</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-2" data-title="PELE Local Exploration of the different substrates in FraCm1 and FraCm2."><figure><figcaption><b id="Fig2" class="c-article-section__figure-caption" data-test="figure-caption-text">Fig. 2: PELE Local Exploration of the different substrates in FraC<sub>m1</sub> and FraC<sub>m2</sub>.</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/2" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig2_HTML.png?as=webp"><img aria-describedby="Fig2" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig2_HTML.png" alt="figure 2" loading="lazy" width="685" height="278"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-2-desc"><p><b>a</b>, Correlation between computational PELE normalized relative activity (number of catalytic PELE poses/total number of accepted PELE steps and number of ester groups in the substrate) and experimental ester-hydrolysing activity (‘Exp. activity’ in the figure) for glyceryl tripropionate (labelled as GRP), vinyl acetate (VIN), phenyl acetate (PAE), <b>TE</b> and <b>ETE</b>. <b>b</b>, Schematic representation of PELE Local Exploration with <b>ETE</b>. Arrows indicate the random movement of the substrate in an N-terminal Å box during the PELE local sampling simulation. Calculations and raw data provided in Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">1</a>.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/2" data-track-dest="link:Figure2 Full size image" aria-label="Full size image figure 2" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><p>The stability of both the soluble form and the N-terminal region inserted into a membrane model by molecular dynamics (MD) was further confirmed (Supplementary Figs. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">3</a>–<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">6</a>). The analysis of catalytic distances for the inserted models indicated adequate hydrogen bond values, with the histidine–serine distance along the simulation being lower for the N-terminal mutant FraC<sub>m1</sub> (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig3">3a–d</a>) than for the globular mutant FraC<sub>m2</sub> (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig3">3e–h</a>).</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-3" data-title="Membrane MD simulation of FraCm1 and FraCm2."><figure><figcaption><b id="Fig3" class="c-article-section__figure-caption" data-test="figure-caption-text">Fig. 3: Membrane MD simulation of FraC<sub>m1</sub> and FraC<sub>m2</sub>.</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/3" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig3_HTML.png?as=webp"><img aria-describedby="Fig3" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig3_HTML.png" alt="figure 3" loading="lazy" width="685" height="609"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-3-desc"><p>FraC<sub>m1</sub>: <b>a</b>, N-terminal fragments are shown from residues 13 to 28, where the catalytic active site is located. Ser21 (introduced by a Thr21Ser mutation), His20 (introduced by a Lys20His mutation) and Glu24 (detected in the wild-type protein) formed the catalytic triad, and Asp17 and Glu24 (both detected in the wild-type protein) could act as the acid residue of the catalytic triad due to the proximity of the histidine residue. Hydrogen bond interactions between catalytic residues and their distances are displayed. <b>b</b>,<b>c</b>, Histogram (<b>b</b>) and violin plot (<b>c</b>) of the distances between two catalytic residues in the catalytic triad throughout the simulation. <b>d</b>, Distance between two catalytic residues over the simulation time. FraC<sub>m2</sub>: <b>e</b>, The active site in the globular region is shown. Ser38 (introduced by an Asp38Ser mutation), His175 (detected in the wild-type protein) and Glu40 (detected in the wild-type protein) conformed to the catalytic triad, and Gln173 (introduced by a Glu173Gln mutation) in the vicinity of the identified pocket conformed to the oxyanion hole. <b>f</b>,<b>g</b>, Histogram (<b>f</b>) and violin plot (<b>g</b>) of the distances between two catalytic residues in the catalytic triad throughout the simulation. <b>h</b>, Distance between two catalytic residues over the simulation time. MD simulations were collected and analysed with GROMACS (version 5.1.2) and OPENMM (version 7.3). Calculations and raw data are provided in Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">1</a>.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/3" data-track-dest="link:Figure3 Full size image" aria-label="Full size image figure 3" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><p>We further evaluated whether the two active centres, located inside the nanopores, could be accessed by nPET particles. Although the flexible nature of a single chain cannot directly be translated to that of a PET particle, we explored the flexibility of a PET chain of 200 units (which could mimic the molecular weights of various common PET samples, from 40 to 80 kDa) with MD simulations. The simulation shows that the PET chains in the nPET particle are dynamic entities and that their shape changes considerably over time (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig4">4a</a>), introducing multiple protuberances that could fit the pore dimensions, as observed in simple molecular docking (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig4">4b</a>). Moreover, one would expect that these protuberances could be stabilized by the pore shape (in an induced fit manner) and increase with partial hydrolysis. Moreover, one could also anticipate these particles to be formed from various chains, having multiple loose ends.</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-4" data-title="Molecular dynamic simulation of PET in solution."><figure><figcaption><b id="Fig4" class="c-article-section__figure-caption" data-test="figure-caption-text">Fig. 4: Molecular dynamic simulation of PET in solution.</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/4" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig4_HTML.png?as=webp"><img aria-describedby="Fig4" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig4_HTML.png" alt="figure 4" loading="lazy" width="685" height="713"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-4-desc"><p><b>a</b>, Solvent-accessible surface area of a nPET particle of 200 monomers over ~50 ns and 40 °C. Snapshots from the simulation are shown for different frames (where a frame is 0.1 ns). <b>b</b>, Graphical representation of a schematic coupling between the pore and the polymer. Calculations and raw data are provided in Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">1</a>.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/4" data-track-dest="link:Figure4 Full size image" aria-label="Full size image figure 4" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><h3 class="c-article__sub-heading" id="Sec4">npFraC<sub>m1</sub> and npFraC<sub>m2</sub> catalyse ester hydrolysis of TE and/or ETE</h3><p>FraC<sub>WT</sub>, FraC<sub>m1</sub> and FraC<sub>m2</sub> were produced in an <i>Escherichia coli</i> expression system and purified (at least 95% pure; Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">7</a> and <a data-track="click" data-track-label="link" data-track-action="section anchor" href="/articles/s41929-023-01048-6#Sec21">Source Data</a>). After evaluating that the intrinsic features and activities of the two mutants were indistinguishable from those of the wild type (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">8a–d</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">2</a>), they were assembled as individual octameric pore water-soluble particles by adding them to empty nanodiscs to mimic the real situation encountered by actinoporins<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 21" title="Tanaka, K., Caaveiro, J. M. M., Morante, K., González-Mañas, J. M. & Tsumoto, K. Structural basis for self-assembly of a cytolytic pore lined by protein and lipid. Nat. Commun. 6, 1–11 (2015)." href="/articles/s41929-023-01048-6#ref-CR21" id="ref-link-section-d10881139e1331">21</a></sup> (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">9</a> and <a data-track="click" data-track-label="link" data-track-action="section anchor" href="/articles/s41929-023-01048-6#Sec21">Source Data</a>). See details in Supplementary Note <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">2</a>. These preparations, referred to as npFraC<sub>WT</sub>, npFraC<sub>m1</sub> and npFraC<sub>m2</sub>, were tested for their hydrolytic activity (Supplementary Note <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">3</a>).</p><p>The measurement of <i>p</i>-nitrophenol (<i>p</i>-NP) release showed that npFraC<sub>m1</sub> and npFraC<sub>m2</sub> were capable of hydrolysing the three <i>p</i>-NP esters tested (pH 7.0, 40 °C), which are model esters for serine ester hydrolases (Extended Data Table <a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/s41929-023-01048-6#Tab2">1</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">3</a>). Both engineered nanopores hydrolysed (pH 7.0, 40 °C) also the three model esters used for PELE exploration, <b>ETE</b> and <b>TE</b>, although npFraC<sub>m1</sub> showed low specific activity for <b>TE</b> (Extended Data Table <a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/s41929-023-01048-6#Tab2">1</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">3</a>). We observed notable correlations between the computed PELE-normalized relative catalytic activity and the experimental ester-hydrolysing activity (<i>R</i><sup>2</sup> = 0.72 for npFraC<sub>m1</sub>; <i>R</i><sup>2</sup> = 0.566 for npFraC<sub>m2</sub>; Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig2">2a</a>). npFraC<sub>m1</sub> and npFraC<sub>m2</sub> hydrolysed <b>ETE</b> with <i>k</i><sub>cat</sub>/<i>K</i><sub>m</sub> values (Table <a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/s41929-023-01048-6#Tab1">1</a>, Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">10</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">4</a>) that were directly benchmarked with those reported for PETases (550–17,000 M<sup>−1</sup> s<sup>−</sup><sup>1</sup>)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 31" title="Bååth, J. A., Borch, K., Jensen, K., Brask, J. & Westh, P. Comparative biochemistry of four polyester (PET) hydrolases. ChemBioChem 22, 1627–1637 (2021)." href="/articles/s41929-023-01048-6#ref-CR31" id="ref-link-section-d10881139e1442">31</a></sup>.</p><div class="c-article-table" data-test="inline-table" data-container-section="table" id="table-1"><figure><figcaption class="c-article-table__figcaption"><b id="Tab1" data-test="table-caption">Table 1 Kinetic hydrolysis parameters of ETE and nPET<sub>b</sub> for npFraC<sub>m1</sub> and npFraC<sub>m2</sub></b></figcaption><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="table-link" data-track="click" data-track-action="view table" data-track-label="button" rel="nofollow" href="/articles/s41929-023-01048-6/tables/1" aria-label="Full size table 1"><span>Full size table</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><p>Using <i>p</i>-NP propionate as a substrate under different conditions, npFraC<sub>m1</sub> and npFraC<sub>m2</sub> showed maximum hydrolytic activity at a pH of approximately 9.0 and temperatures from 35 °C to 45 °C (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">11</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">5</a>). This is strongly consistent with the experimentally determined thermal melting temperature of FraC<sub>WT</sub> (<i>T</i><sub>m</sub> = 55.3 ± 0.5 °C), FraC<sub>m1</sub> (<i>T</i><sub>m</sub> = 57.4 ± 0.5 °C) and FraC<sub>m2</sub> (<i>T</i><sub>m</sub> = 61.6 ± 0.5 °C), as monitored by circular dichroism spectroscopy at 220 nm (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">12</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">6</a>).</p><h3 class="c-article__sub-heading" id="Sec5">Deconstruction of nPET by npFraC<sub>m1/m2</sub> </h3><p>We further evaluated the possibility that npFraC<sub>m1</sub> and npFraC<sub>m2</sub> not only hydrolyse <b>TE</b> and <b>ETE</b> but also break down nPET particles of different types across the pores. Instead of using naturally occurring nPET particles, for example, those in wastewater treatment plants<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 18" title="Cui, Y. et al. Computational redesign of a PETase for plastic biodegradation under ambient condition by the GRAPE strategy. ACS Catal. 11, 1340–1350 (2021)." href="/articles/s41929-023-01048-6#ref-CR18" id="ref-link-section-d10881139e2015">18</a></sup>, we chose to prepare, in vitro, particles of different nature. This allows us to assess their physical–chemical characteristics and their alterations during the degradation process, and the released products. nPET particles were produced (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">Supplementary Methods</a>) from PET from a bottle of daily life (from a local shop, Granini brand, nPET<sub>b</sub>) and from a commodity thermoplastic polymer resin that is widely used for packaging (nonreheat PET resin RAMAPET N180, nPET<sub>c</sub>), as well as from two PET products from GoodFellow Cambridge: biaxially oriented crystalline PET film (GoodFellow crystalline, nPET<sub>GFc</sub>) and amorphous PET film (GoodFellow amorphous, nPET<sub>GFa</sub>). According to the manufacturers, PET<sub>c</sub>, PET<sub>GFa</sub> and PET<sub>GFc</sub> raw materials are of high purity (purity 99.9%), and the composition of PET<sub>b</sub> is according to legislation (Supplementary Note <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">4</a>).</p><p>Using dynamic light scattering (DLS), the <i>Z</i>-average particle size (diameter) measured at 25 °C confirmed the presence of nPET particles with a stable, typical<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Cui, Y. et al. Computational redesign of a PETase for plastic biodegradation under ambient condition by the GRAPE strategy. ACS Catal. 11, 1340–1350 (2021)." href="#ref-CR18" id="ref-link-section-d10881139e2049">18</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Barth, M. et al. Effect of hydrolysis products on the enzymatic degradation of polyethylene terephthalate nanoparticles by a polyester hydrolase from Thermobifida fusca. Biochem. Eng. J. 93, 222–228 (2015)." href="#ref-CR19" id="ref-link-section-d10881139e2049_1">19</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 20" title="Vogel, K. et al. Enzymatic degradation of polyethylene terephthalate nanoplastics analyzed in real time by isothermal titration calorimetry. Sci. Total Environ. 773, 145111 (2021)." href="/articles/s41929-023-01048-6#ref-CR20" id="ref-link-section-d10881139e2052">20</a></sup> size distribution from 53.1 ± 0.1 nm to 108.0 ± 0.1 nm (Fig. 5a, Extended Data Table <a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/s41929-023-01048-6#Tab3">2</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">7</a>), which is in the sub-micro to nanometre range. By differential scanning calorimetry, the degree of crystallinity (%), <i>T</i><sub>G</sub>, melting temperature (<i>T</i><sub>m</sub>), cold crystallization temperature (<i>T</i><sub>c</sub>) and cold crystallization energy (Δ<i>H</i><sub>c</sub>) of the bulk PET materials and the nPET particles could be also obtained (Extended Data Table <a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/s41929-023-01048-6#Tab3">2</a> and Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">13</a>). The crystallinity, <i>T</i><sub>G</sub>, <i>T</i><sub>m</sub> and <i>T</i><sub>c</sub> ranged from 1.3% to 34.5%, from 72.0 °C to 75.7 °C, from 245.9 °C to 251.9 °C, and from 192.8 °C to 215.1 °C, respectively, which is in the range of reported datasets<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 12" title="Wei, R. et al. Mechanism-based design of efficient PET hydrolases. ACS Catal. 12, 3382–3396 (2022)." href="/articles/s41929-023-01048-6#ref-CR12" id="ref-link-section-d10881139e2099">12</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 32" title="Menzel, T. et al. Impact of enzymatic degradation on the material properties of poly(ethylene terephthalate). Polymers 13, 3885 (2021)." href="/articles/s41929-023-01048-6#ref-CR32" id="ref-link-section-d10881139e2102">32</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 33" title="Bashir, Z., Al-Aloush, I., Al-Raqibah, I. & Ibrahim, M. Evaluation of three methods for the measurement of crystallinity of pet resins, preforms, and bottles. Polym. Eng. Sci. 40, 2442–2455 (2000)." href="/articles/s41929-023-01048-6#ref-CR33" id="ref-link-section-d10881139e2105">33</a></sup>. For additional considerations, see Supplementary Note <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">5</a>.</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-5" data-title="Particle size distribution of nPETGFa/GFc/b/c as determined by DLS."><figure><figcaption><b id="Fig5" class="c-article-section__figure-caption" data-test="figure-caption-text">Fig. 5: Particle size distribution of nPET<sub>GFa/GFc/b/c</sub> as determined by DLS.</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/5" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig5_HTML.png?as=webp"><img aria-describedby="Fig5" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig5_HTML.png" alt="figure 5" loading="lazy" width="685" height="579"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-5-desc"><p><b>a</b>,<b>b</b>, The distribution mean (Dv,50) size (±s.d. error bar) calculated by the scattered intensity for each particle size from three measurements (<i>n</i> = 3) is represented (<b>a</b>, for nPET<sub><i>x</i></sub> where <i>x</i> refers to GFa/GFc/b/c; <b>b</b>, for nPET<sub>b</sub> obtained at various transfer rates). Particles were obtained by predissolving PET<sub>b</sub> for 2 h at 150 rpm and 25 °C in 1,1,1,3,3,3-hexafluor-2-propanole (from Merck Life Science) and further transferred with the help of a burette into an ice-cold water-containing beaker (in an ice bath), which was strongly agitated (250 rpm). For <b>a</b>, the flow rate was fixed at 1 ml min<sup>−</sup><sup>1</sup>, and for <b>b</b>, it was fixed at 0.05, 0.10, 0.25, 0.5, 1.0, 1.5 and 2.0 ml min<sup>−</sup><sup>1</sup> for producing 69.1, 78.8, 79.5, 85.4, 108.0, 125.9, 126.2 and 153.8 nm particles, respectively (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">Supplementary Methods</a>). DLS datasets were collected at 25 °C with a Malvern Instrument (MALVERN ZETASIER NANO S ZEN 1600, IESMAT) with Dispersion Technology software 4.20 (Malvern Instrument). <b>c</b>, Evaluation of the degradation of nPET<sub>b</sub> particles of different sizes using npFraC<sub>m1/m2</sub>. Reaction conditions: [npFraC<sub>m1/m2</sub>], 1.5 μg ml<sup>−</sup><sup>1</sup>; [nPET<sub>b</sub>], 1.1 mg ml<sup>−</sup><sup>1</sup>; reaction volume, 100 μl; <i>T</i>, 40 °C; pH, 7.0 (20 mM HEPES buffer); reaction time, 48 h. The reactions were maintained in 2-ml safe-lock Eppendorf polypropylene tubes (ref. 0030 120.094) in a thermoshaker (model Thermomixer comfort, Eppendorf AG) at 1,000 rpm. The reactions were stopped by adding 900 μl dimethyl sulfoxide (from Merck Life Science), and the degradation products were immediately analysed by HPLC. Datasets were collected with a Varian Star LC workstation 6.41 (Varian). Quantifications of degradation products were performed by HPLC on the basis of calibrations with purified standards. Values in <b>a</b> and <b>b</b> are plotted as the mean of three independent replicates (<i>n</i> = 3) with the reported error ranges and s.d. calculated using Dispersion Technology software 4.20 (Malvern Instrument). Values in <b>c</b> and <b>d</b> are plotted as the mean of three independent replicates (<i>n</i> = 3) with the reported error ranges and s.d. calculated using the STDEV.S function in Excel 2019 (calculations and raw data are provided in <a data-track="click" data-track-label="link" data-track-action="section anchor" href="/articles/s41929-023-01048-6#Sec21">Source Data</a>). The figure was made using Excel 2019. Raw data are shown in Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">7</a>.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/5" data-track-dest="link:Figure5 Full size image" aria-label="Full size image figure 5" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><p>npFraC<sub>m1</sub> or npFraC<sub>m2</sub> were added at a concentration of 1.5 μg ml<sup>−1</sup> (or 76 nM; Supplementary Note <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">6</a>) to buffered suspensions (pH 7.0) containing 2.23 ± 0.06 mg ml<sup>−1</sup> nPET<sub>GFa</sub>, nPET<sub>GFc</sub>, nPET<sub>b</sub> and nPET<sub>c</sub>, and the degradation products were observed by high-performance liquid chromatography (HPLC) (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig6">6a,b</a>) after 48 h of incubation at <i>T</i><sub>opt</sub>, 40 °C. <i>T</i><sub>opt</sub> refers to the temperature at which maximal ester-hydrolytic activity was observed using <i>p</i>-NP propionate (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">11</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">5</a>); at this temperature, npFraC<sub>m1</sub> and npFraC<sub>m2</sub> retained 54.7 ± 2.1% and 32.7 ± 6.3%, respectively, of their original activities after 48 h (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">14</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">8</a>). The chemical structures of all degradation products identified under our assay conditions are detailed in Extended Data Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig8">1</a>. The identity was confirmed by high-resolution mass spectrometry (MS) analysis using products purified from reaction mixtures by semipreparative HPLC (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">15</a>).</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-6" data-title="Degradation profiles of nPETGFa/GFc/b/c treated with npFraCm1/m2."><figure><figcaption><b id="Fig6" class="c-article-section__figure-caption" data-test="figure-caption-text">Fig. 6: Degradation profiles of nPET<sub>GFa/GFc/b/c</sub> treated with npFraC<sub>m1/m2</sub>.</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/6" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig6_HTML.png?as=webp"><img aria-describedby="Fig6" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig6_HTML.png" alt="figure 6" loading="lazy" width="685" height="542"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-6-desc"><p><b>a</b>,<b>b</b>, HPLC profiles of degradation products released by npFraC<sub>m1</sub> (<b>a</b>) or npFraC<sub>m2</sub> (<b>b</b>) under the experimental conditions: [npFraC<sub>m1/m2</sub>], 1.5 μg ml<sup>−</sup><sup>1</sup>; [nPET<sub>GFa/GFc/b/c</sub>], 1.1 mg ml<sup>−</sup><sup>1</sup>; reaction volume, 1,000 μl; <i>T</i>, 40 °C; pH, 7.0 (20 mM HEPES buffer); reaction time, 48 h. The reactions were maintained in 2-ml safe-lock Eppendorf polypropylene tubes (ref. 0030 120.094) in a thermoshaker (model Thermomixer comfort, Eppendorf AG) at 1,000 rpm. Aliquots of 100 μl were obtained, the reactions were stopped by adding 900 μl dimethyl sulfoxide (from Merck Life Science) and the degradation products were immediately analysed by HPLC. Datasets were collected with a Varian Star LC workstation 6.41 (Varian). All reactions and analyses were performed in triplicate (<i>n</i> = 3), with a representative chromatogram per enzyme and nPET particle shown. <b>c</b>,<b>d</b>, Concentration of degradation products found to be present in reaction mixtures with npFraC<sub>m1</sub> (<b>c</b>) or npFraC<sub>m2</sub> (<b>d</b>). Quantification was performed for products shown in <b>a</b> and <b>b</b> with unambiguous identification and for which enough material was recovered for HPLC calibration, namely from <b>T</b> to <b>ETETE</b> (Extended Data Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig8">1</a>); purification was performed by semipreparative HPLC, and the molecular weights and structures were determined by mass spectrometry (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">15</a>). Values are plotted as the mean of three independent replicates (<i>n</i> = 3) with the reported error ranges and s.d. calculated using the STDEV.S function in Excel 2019 (calculations and raw data are provided in <a data-track="click" data-track-label="link" data-track-action="section anchor" href="/articles/s41929-023-01048-6#Sec21">Source Data</a>). The figure was constructed using Excel 2019. Raw data are shown in Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">9</a>.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/6" data-track-dest="link:Figure6 Full size image" aria-label="Full size image figure 6" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><p>The HPLC chromatograms in Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig6">6a</a> revealed four main degradation products when nPET particles were treated with npFraC<sub>m1</sub>: <b>ETE</b>, <b>ETETE</b>, <b>TE</b> and <b>TETETE</b>, in this order, with <b>T</b> being below the detection limit under our assay conditions. This substrate profile was markedly different from that of npFraC<sub>m2</sub>, which yields a higher diversity of oligomers, herein referred to as <b>TET</b>, <b>TETE</b>, <b>TETET</b> and <b>TETETE</b>, in addition to <b>T</b>, <b>TE</b> and <b>ETE</b> (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig6">6b</a>). Under the same experimental conditions, <i>Is</i>PETase, LCC<sub>WT</sub> and LCC<sub>WCCG</sub> released <b>T</b>, <b>TE</b>, <b>ETE</b> and the oligomers <b>TET</b>, <b>TETE</b> and/or <b>TETETE</b>, with <b>T</b> and <b>TE</b> being the main degradation products (Extended Data Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig9">2a–c</a>); the presence of these products is in accordance with the literature and reaction mechanisms<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 15" title="Tournier, V. et al. An engineered PET depolymerase to break down and recycle plastic bottles. Nature 580, 216–219 (2020)." href="/articles/s41929-023-01048-6#ref-CR15" id="ref-link-section-d10881139e2476">15</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 34" title="Joo, S. et al. Structural insight into molecular mechanism of poly(ethylene terephthalate) degradation. Nat. Commun. 9, 382 (2018)." href="/articles/s41929-023-01048-6#ref-CR34" id="ref-link-section-d10881139e2479">34</a></sup>. However, <b>TET</b> was not detected or suggested as a potential degradation product during the degradation of PET film by <i>Is</i>PETase<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 34" title="Joo, S. et al. Structural insight into molecular mechanism of poly(ethylene terephthalate) degradation. Nat. Commun. 9, 382 (2018)." href="/articles/s41929-023-01048-6#ref-CR34" id="ref-link-section-d10881139e2490">34</a></sup>.</p><p>According to calibration with pure standards (<b>T</b>, <b>TE</b>, <b>ETE</b>, <b>TET</b>, <b>TETE</b> and <b>ETETE</b>), the concentration of degradation products at 48 h using npFraC<sub>m1</sub> ranged from 1,718 to 5,743 µM, depending on the type of nPET (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig6">6c</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">9</a>), with nPET<sub>GFa</sub> and nPET<sub>GFc</sub> being degraded to a higher extent than nPET<sub>b</sub> and nPET<sub>c</sub>, despite their higher crystallinity. In all cases, <b>ETE</b> was the major degradation product, whose concentration ranged from 2.1- to 12.5-fold higher than that of <b>TE</b>, depending on the type of nPET. When npFraC<sub>m2</sub> was used, the concentration of degradation products at 48 h ranged from 1,500 to 2,660 µM, depending on the type of nPET, with nPET<sub>b</sub> and nPET<sub>GFa</sub> being the preferred particles, and <b>TE</b> was produced at approximately 3-fold higher concentrations than <b>ETE</b> (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig6">6d</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">9</a>). <i>Is</i>PETase, LCC<sub>WT</sub> and LCC<sub>WCCG</sub> released degradation products at concentrations of 1,154–1,884 µM, 312–410 µM and 309–414 µM, respectively, depending on the type of nPET (Extended Data Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig9">2d–f</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">9</a>). In all cases, <b>T</b> was observed, and <b>TE</b> was found to have a concentration 6- to 31-fold higher than <b>ETE</b>, depending on the nPET and the PETase. The higher efficiency of <i>Is</i>PETase compared with LCC<sub>WT</sub> and LCC<sub>WCCG</sub> could be associated with the higher activity of this enzyme at 40 °C compared with LCC variants<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 17" title="Yoshida, S. et al. A bacterium that degrades and assimilates poly(ethylene terephthalate). Science 351, 1196–1199 (2016)." href="/articles/s41929-023-01048-6#ref-CR17" id="ref-link-section-d10881139e2590">17</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 18" title="Cui, Y. et al. Computational redesign of a PETase for plastic biodegradation under ambient condition by the GRAPE strategy. ACS Catal. 11, 1340–1350 (2021)." href="/articles/s41929-023-01048-6#ref-CR18" id="ref-link-section-d10881139e2593">18</a></sup>.</p><p>This finding confirmed the capacity of npFraC<sub>m1</sub> and npFraC<sub>m2</sub> to efficiently deconstruct nPET materials at 40 °C. The uniqueness of npFraC<sub>m1</sub> compared with the npFraC<sub>m2</sub>, <i>Is</i>PETase and LCC<sub>WT/WCCG</sub> variants in degrading nPET is that <b>ETE</b> is the main degradation product compared with <b>TE</b>, with no appreciable formation of <b>T</b> under our assay conditions. This feature could represent an advantage over the other enzymatic systems that will produce a mixture of <b>E</b>, <b>T</b>, <b>TE</b> and <b>ETE</b>.</p><h3 class="c-article__sub-heading" id="Sec6">Time course and kinetic study of nPET<sub>b</sub> particle degradation</h3><p>The time course of the released degradation products, determined by HPLC using npFraC<sub>m1</sub> and npFraC<sub>m2</sub>, was followed and compared with that of LCC<sub>WT</sub> at 40 °C and pH 7.0. nPET<sub>b</sub> (diameter 108.0 ± 0.1 nm; Extended Data Table <a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/s41929-023-01048-6#Tab3">2</a>) was targeted as the closest example to real nPET substrates, and all three enzymatic preparations were able to efficiently degrade it. Although, <i>Is</i>PETase is currently the most active wild-type enzyme for PET hydrolysis at around 40 °C, LCC<sub>WT</sub> was selected for comparative purposes as a benchmark PETase given its higher stability compared with mesophilic <i>Is</i>PETase under our thermal assay conditions, of 40 °C (for details, see Supplementary Note <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">7</a>; Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">16</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">11</a>).</p><p>As shown in Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig7">7</a> (Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">10</a>), degradation products were constantly observed since the early stages for the three enzymatic preparations. Under the tested assay conditions, the time course of degradation product release was much steeper with npFraC<sub>m2</sub> than with npFraC<sub>m1</sub> in the first few reaction hours. Whereas npFraC<sub>m2</sub> was only highly active until 5–6 h of incubation, hydrolysis by npFraC<sub>m1</sub> continuously progresses up to 24 h where almost maximal depolymerization was achieved, as for LCC<sub>WT</sub> for which hydrolysis progressed beyond 24 h. This is consistent with the lower thermal stability of npFraC<sub>m2</sub> (<i>t</i><sub>1/2</sub> at 40 °C: 5.3 ± 0.2 h) compared with npFraC<sub>m1</sub> (<i>t</i><sub>1/2</sub> at 40 °C > 48 h) under the assay conditions (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">14</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">8</a>). Additionally, the better positioning of the long protuberances that conformed the nPET particles (<a data-track="click" data-track-label="link" data-track-action="section anchor" href="/articles/s41929-023-01048-6#Sec8">Discussion</a>) in the nanopore channel once the particle is seated at the pore entrance, could also explain the differences in the reaction kinetics and dynamics of npFraC<sub>m1</sub> and npFraC<sub>m2</sub>. Regardless of the underlying reasons for a different kinetics, these data demonstrate that the access of nPET particles across the nanopores is not a limiting factor during depolymerization mediated by npFraC<sub>m1</sub> and npFraC<sub>m2</sub>.</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-7" data-title="Time course study of nPETb degradation using npFraCm1/m2 compared with LCCWT."><figure><figcaption><b id="Fig7" class="c-article-section__figure-caption" data-test="figure-caption-text">Fig. 7: Time course study of nPET<sub>b</sub> degradation using npFraC<sub>m1/m2</sub> compared with LCC<sub>WT</sub>.</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/7" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig7_HTML.png?as=webp"><img aria-describedby="Fig7" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig7_HTML.png" alt="figure 7" loading="lazy" width="685" height="187"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-7-desc"><p><b>a</b>–<b>c</b>, Time course degradation of nPET<sub>b</sub> particles using npFraC<sub>m1</sub> (<b>a</b>), npFraC<sub>m2</sub> (<b>b</b>) and LCC<sub>WT</sub> (<b>c</b>). Reaction conditions and dataset collections are detailed in Figs. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig5">5</a> and <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig6">6</a>, and Extended Data Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig9">2</a>. In brief: 1.1 mg ml<sup>−</sup><sup>1</sup> nPET<sub>b</sub>, 1.5 μg ml<sup>−</sup><sup>1</sup> npFraC<sub>m1/m2</sub>, LCC<sub>WT</sub>, 40 °C, pH 7.0. Values are plotted as the mean of three independent replicates (<i>n</i> = 3) with the reported error ranges and s.d. calculated using the STDEV.S function in Excel 2019 (calculations and raw data are provided in <a data-track="click" data-track-label="link" data-track-action="section anchor" href="/articles/s41929-023-01048-6#Sec21">Source Data</a>). Raw data are provided in Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">10</a>.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/s41929-023-01048-6/figures/7" data-track-dest="link:Figure7 Full size image" aria-label="Full size image figure 7" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><p>To evaluate the effect of the particle size, we prepared nPET<sub>b</sub> particles of different mean sizes (diameter), namely from 69.1 ± 0.8 to 153.8 ± 1.4 nm (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig5">5b</a> and Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">7</a>). We discovered that smaller particles were hydrolysed by npFraC<sub>m1</sub> more easily than larger particles (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig5">5c</a>), whereas npFraC<sub>m2</sub> preferred particles from 85.4 to 108 nm (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig5">5d</a>), in accordance with the increased exposure of its active site to the particles (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig4">4</a>). These data suggest that, while particles as large as 153.8 nm were degraded (from 8.0- to 4.4-fold lower than that of the best performing size), there will be a particle size limit that will most likely no longer be degraded. Regardless of this finding, the analysed particle sizes are well above the npFraC<sub>m1</sub> and npFraC<sub>m2</sub> sizes, namely 6.7 nm in the <i>cis</i> entry and 1.9 nm in the <i>trans</i> exit<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 21" title="Tanaka, K., Caaveiro, J. M. M., Morante, K., González-Mañas, J. M. & Tsumoto, K. Structural basis for self-assembly of a cytolytic pore lined by protein and lipid. Nat. Commun. 6, 1–11 (2015)." href="/articles/s41929-023-01048-6#ref-CR21" id="ref-link-section-d10881139e2845">21</a></sup> (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig4">4</a>).</p><p>The results of the conventional (<sup>conv</sup>MM) and inverse (<sup>inv</sup>MM) Michaelis‒Menten kinetics<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 35" title="Erickson, E. et al. Comparative performance of PETase as a function of reaction conditions, substrate properties, and product accumulation. ChemSusChem 15, e202101932 (2022)." href="/articles/s41929-023-01048-6#ref-CR35" id="ref-link-section-d10881139e2859">35</a></sup> are summarized in Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">17a–f</a> (Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">12</a> and <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">13</a>), and the parameters resulting from the fits (pH 7.0, 40 °C) are presented in Table <a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/s41929-023-01048-6#Tab1">1</a>. The results highlight that the affinity, deconstruction rates and reactive site density (<i>Γ</i><sub>attack</sub>) of the catalytic nanopores are comparable to those of soluble PETases, demonstrating that having a catalytic unit in a nanopore does not substantially affect either the affinity for, or the reaction rate towards, the nPET particles, as well as the concentration of reactive sites available per gram of substrate compared with a soluble enzyme with free access to the particles. For additional kinetic details, see Supplementary Note <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">7</a>.</p><h3 class="c-article__sub-heading" id="Sec7">Morphological evolution during nPET<sub>b</sub> degradation by npFraC<sub>m1</sub> and npFraC<sub>m2</sub> </h3><p>The change in particle size distribution and morphology was analysed by field emission scanning electron microscopy (FE-SEM) in dried samples collected before, during and after hydrolysis. The FE-SEM micrographs in Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">18</a> (<a data-track="click" data-track-label="link" data-track-action="section anchor" href="/articles/s41929-023-01048-6#Sec21">Source Data</a>) show the morphological degradation of the nPET<sub>b</sub> particles by npFraC<sub>m1</sub> and npFraC<sub>m2</sub>. The as-synthesized spheres, with a relatively uniform size distribution, start aggregating and degrading after few minutes of hydrolysis, appearing covered by a film of insoluble degradation products in the samples incubated with npFraC<sub>m1/m2</sub>. Eventually, all particles disappeared and only large aggregates of degradation products are visible, as observed for PETases with a random-hydrolysis mechanism<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 20" title="Vogel, K. et al. Enzymatic degradation of polyethylene terephthalate nanoplastics analyzed in real time by isothermal titration calorimetry. Sci. Total Environ. 773, 145111 (2021)." href="/articles/s41929-023-01048-6#ref-CR20" id="ref-link-section-d10881139e2914">20</a></sup>.</p></div></div></section><section data-title="Discussion"><div class="c-article-section" id="Sec8-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec8">Discussion</h2><div class="c-article-section__content" id="Sec8-content"><p>This study focused on the design of two pore-based biocatalytic nanoreactors for nPET particle depolymerization, using a non-catalytic pore-forming protein as the starting target. Through a combination of computational structure-based modelling tools and different experimental techniques, we provide robust datasets demonstrating that these biocatalytic nanopores can break down primary nPET of different types at 40 °C. The diameter of the nPET particles produced and tested ranged from approximately 53 to 154 nm, whereas the FraC pore has an inner diameter ranging from 1.9 nm (<i>cis</i> exit) to 6.7 nm (<i>trans</i> exit)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 21" title="Tanaka, K., Caaveiro, J. M. M., Morante, K., González-Mañas, J. M. & Tsumoto, K. Structural basis for self-assembly of a cytolytic pore lined by protein and lipid. Nat. Commun. 6, 1–11 (2015)." href="/articles/s41929-023-01048-6#ref-CR21" id="ref-link-section-d10881139e2933">21</a></sup>. Therefore, if we stick with this static image, one might suggest that nPET particles would not be able to access the artificial active centres introduced into the nanopores, although experimental evidence shows otherwise. Since the entry of each particle in its entirety into the pore would not be possible, we propose that the nPET particles may have protuberances and that it is not the particles that enter into the nanopores but the protuberances (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig4">4</a>), the size of which will be reduced during hydrolysis, and with it the size of the particles, that will be more accessible as the hydrolysis proceeds. This may agree with the fact that the molecular weight of sub-micro- and nano-sized PET particles may be lower compared with that of the pristine materials, and that the degradability of sub-micrometre particles and nanoparticles may not only be due to the increased surface area, but also to the lower chain lengths and more flexible polymer chain ends<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 36" title="Pfaff, L. et al. Multiple substrate binding mode-guided engineering of a thermophilic PET hydrolase. ACS Catal. 12, 9790–9800 (2022)." href="/articles/s41929-023-01048-6#ref-CR36" id="ref-link-section-d10881139e2940">36</a></sup>.</p><p>The data suggest that npFraC<sub>m1</sub> performs endo-hydrolysis on the small protuberances that can access the internal area of the catalytic nanopore; this would result in the generation of <b>E</b> (<sup><b>E</b></sup>PET) and <b>T</b> (or <sup><b>T</b></sup>PET) terminal PET oligomers, from which smaller subproducts, preferentially <b>ETE</b> and <b>ETETE</b> (from <sup><b>E</b></sup>PET), and to a lesser extent <b>TETETE</b> (from <sup><b>T</b></sup>PET), will be formed by exo-type chain scission. In the case of npFraC<sub>m2</sub>, the active site is remarkably more exposed to the solvent and to different accommodations of the sub-micro- and nanoparticles (and of its protuberances), initiating a more random endo- and exo-type chain degradation that yields a high diversity of oligomers and monomers. The high accumulation of all such degradation products (Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig6">6</a>) suggests that the cleavage of <sup><b>T</b></sup>PET and <sup><b>E</b></sup>PET by npFraC<sub>m2</sub> would occur at similar rates. This finding contrasts with the PET degradation process proposed for <i>Is</i>PETase, where the terminal digestion step of <sup><b>T</b></sup>PET is preferred<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 34" title="Joo, S. et al. Structural insight into molecular mechanism of poly(ethylene terephthalate) degradation. Nat. Commun. 9, 382 (2018)." href="/articles/s41929-023-01048-6#ref-CR34" id="ref-link-section-d10881139e3005">34</a></sup>.</p><p>Compared with benchmark PETases, namely LCC<sub>WT</sub>, LCC<sub>WCCG</sub> and <i>Is</i>PETase, we discovered remarkable differences in hydrolysis capacity and degradation products (Extended Data Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig10">3</a>). Under our experimental conditions, npFraC<sub>m1</sub> breaks down nPET to <b>ETE</b> as the main degradation product without appreciable production of <b>T</b>, which is a final PET-building block that is usually produced by the benchmark PETases<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Wei, R. et al. Mechanism-based design of efficient PET hydrolases. ACS Catal. 12, 3382–3396 (2022)." href="#ref-CR12" id="ref-link-section-d10881139e3031">12</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Son, H. F. et al. Rational protein engineering of thermo-stable PETase from Ideonella sakaiensis for highly efficient PET degradation. ACS Catal. 9, 3519–3526 (2019)." href="#ref-CR13" id="ref-link-section-d10881139e3031_1">13</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Bell, E. L. et al. Directed evolution of an efficient and thermostable PET depolymerase. Nat. Catal. 5, 673–681 (2022)." href="#ref-CR14" id="ref-link-section-d10881139e3031_2">14</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Tournier, V. et al. An engineered PET depolymerase to break down and recycle plastic bottles. Nature 580, 216–219 (2020)." href="#ref-CR15" id="ref-link-section-d10881139e3031_3">15</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Lu, H. et al. Machine learning-aided engineering of hydrolases for PET depolymerization. Nature 604, 662–667 (2022)." href="#ref-CR16" id="ref-link-section-d10881139e3031_4">16</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Yoshida, S. et al. A bacterium that degrades and assimilates poly(ethylene terephthalate). Science 351, 1196–1199 (2016)." href="#ref-CR17" id="ref-link-section-d10881139e3031_5">17</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Cui, Y. et al. Computational redesign of a PETase for plastic biodegradation under ambient condition by the GRAPE strategy. ACS Catal. 11, 1340–1350 (2021)." href="#ref-CR18" id="ref-link-section-d10881139e3031_6">18</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" title="Barth, M. et al. Effect of hydrolysis products on the enzymatic degradation of polyethylene terephthalate nanoparticles by a polyester hydrolase from Thermobifida fusca. Biochem. Eng. J. 93, 222–228 (2015)." href="#ref-CR19" id="ref-link-section-d10881139e3031_7">19</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 20" title="Vogel, K. et al. Enzymatic degradation of polyethylene terephthalate nanoplastics analyzed in real time by isothermal titration calorimetry. Sci. Total Environ. 773, 145111 (2021)." href="/articles/s41929-023-01048-6#ref-CR20" id="ref-link-section-d10881139e3034">20</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 34" title="Joo, S. et al. Structural insight into molecular mechanism of poly(ethylene terephthalate) degradation. Nat. Commun. 9, 382 (2018)." href="/articles/s41929-023-01048-6#ref-CR34" id="ref-link-section-d10881139e3037">34</a></sup>. Instead, npFraC<sub>m2</sub> behaves similarly to benchmark PETases in the sense that it can fully convert nPET to <b>T</b>, but unlike them, it produces a greater diversity of degradation products. The possibility of using FraC to design two catalytic nanopores capable of degrading nPET at relatively low temperatures (40 °C) and with different degradation profiles among them and compared with benchmark PETases demonstrates the versatility of this protein as a scaffold for the design of pore-based catalytic nanoreactors to deconstruct nPET particles. Note that up to eight refined artificial biocatalytic sites could be incorporated due to the homo-octamer assembly. Therefore, the limitations in the access of the nPET particles to the active sites can be compensated by the presence of multiple catalytic groups, whose number would be higher than in conventional enzymes with PETase activity. Indeed, our results demonstrated that, using the same conditions and particles, and considering the limitations of access to the particles, the catalytic efficiency for nPET<sub>b</sub> of npFraC<sub>m1</sub> and npFraC<sub>m2</sub> is similar to that of soluble LCC<sub>WT</sub><sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 15" title="Tournier, V. et al. An engineered PET depolymerase to break down and recycle plastic bottles. Nature 580, 216–219 (2020)." href="/articles/s41929-023-01048-6#ref-CR15" id="ref-link-section-d10881139e3054">15</a></sup>.</p><p>The above features, with the easy production, simple purification and stability of the engineered catalytic nanopores designed herein (Supplementary Note <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">2</a>), can create new alternatives to degrade sub-micro- and nano-sized PET, for example, in wastewater treatment plants<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 18" title="Cui, Y. et al. Computational redesign of a PETase for plastic biodegradation under ambient condition by the GRAPE strategy. ACS Catal. 11, 1340–1350 (2021)." href="/articles/s41929-023-01048-6#ref-CR18" id="ref-link-section-d10881139e3064">18</a></sup>, under sustainable conditions, for example, 40 °C. The proof of concept presented in this study may open up new lines of research. We envision, for example, the co-integration of the two artificial hydrolytic sites or designing thermostable variants by further protein engineering efforts to have robustness above 70 °C to degrade real-world plastic pollution at an industrially relevant scale<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 18" title="Cui, Y. et al. Computational redesign of a PETase for plastic biodegradation under ambient condition by the GRAPE strategy. ACS Catal. 11, 1340–1350 (2021)." href="/articles/s41929-023-01048-6#ref-CR18" id="ref-link-section-d10881139e3068">18</a></sup>. Also, the co-integration of engineered pore-forming proteins and nanoscale materials or membranes to develop inorganic–organic hybrid catalysts that can act as both filtering systems to effectively capture sub-micro- and nano-sized PET, mimicking the well-known water desalination nanopores<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 37" title="Heiranian, M., Farimani, A. & Aluru, N. Water desalination with a single-layer MoS2 nanopore. Nat. Commun. 6, 8616 (2015)." href="/articles/s41929-023-01048-6#ref-CR37" id="ref-link-section-d10881139e3072">37</a></sup>, and reaction chambers to further degrade synthetic particles<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 38" title="Gigault, J. et al. Nanoplastics are neither microplastics nor engineered nanoparticles. Nat. Nanotechnol. 16, 501–507 (2021)." href="/articles/s41929-023-01048-6#ref-CR38" id="ref-link-section-d10881139e3076">38</a></sup> (Supplementary Note <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">9</a>). The possibility of producing engineered transmembrane pore-forming proteins, which include FraC or other larger pore-forming proteins that can oligomerize in annular pores of more than 30 monomers and large diameters (for example, the 25–40 nm Perfringolysin O<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 39" title="Johnstone, B. A. et al. Cholesterol-dependent cytolysins: the outstanding questions. IUBMB Life 74, 1169–1179 (2022)." href="/articles/s41929-023-01048-6#ref-CR39" id="ref-link-section-d10881139e3084">39</a></sup>), in target cell membranes may also enable the design of microbial reactors with integrated catalytic pores supporting multiple conversions, yet to be defined, as exemplified here by nPET degradation.</p></div></div></section><section data-title="Methods"><div class="c-article-section" id="Sec9-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec9">Methods</h2><div class="c-article-section__content" id="Sec9-content"><h3 class="c-article__sub-heading" id="Sec10">In silico protein and substrate preparation</h3><p>We selected the homo-octamer biological assembly crystal structure of FraC (4TSY<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 40" title="4TSY: crystal structure of FraC with lipids. NCBI https://www.ncbi.nlm.nih.gov/Structure/pdb/4TSY " href="/articles/s41929-023-01048-6#ref-CR40" id="ref-link-section-d10881139e3100">40</a></sup>) and its monomer soluble structure (3W9P<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 41" title="3W9P: crystal structure of monomeric FraC (second crystal form). NCBI https://www.ncbi.nlm.nih.gov/Structure/pdb/3W9P " href="/articles/s41929-023-01048-6#ref-CR41" id="ref-link-section-d10881139e3104">41</a></sup>). Each system was prepared and optimized at pH 7.5 with a Protein Preparation Wizard<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 42" title="Sastry, G. M., Adzhigirey, M., Day, T., Annabhimoju, R. & Sherman, W. Protein and ligand preparation: parameters, protocols, and influence on virtual screening enrichments. J. Comput. Aided Mol. Des. 27, 221–234 (2013)." href="/articles/s41929-023-01048-6#ref-CR42" id="ref-link-section-d10881139e3108">42</a></sup> from Schrodinger to fix the protonation states and correct alternative positions or other common problems. The three model esters (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">1</a>), <b>TE</b> and <b>ETE</b> were modelled using the OPLS2005 force field<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 43" title="Banks, J. L. et al. Integrated Modeling Program, Applied Chemical Theory (IMPACT). J. Comput. Chem. 26, 1752–1780 (2005)." href="/articles/s41929-023-01048-6#ref-CR43" id="ref-link-section-d10881139e3122">43</a></sup>, and the atomic charges were obtained using single-point energy quantum mechanics calculations with density functional theory using the B3LYP-D3 exchange-correlation and CC-PVTZ basis set.</p><h3 class="c-article__sub-heading" id="Sec11">PELE, adaptive simulations and distance metrics</h3><p>The engineering protocol starts by mapping possible substrate binding sites in the target protein. Interactions between the different ester-type substrates and the inner surface of the assembled pore-forming protein scaffold were thus sampled using PELE software<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 44" title="Lecina, D., Gilabert, J. F. & Guallar, V. Adaptive simulations, towards interactive protein–ligand modeling. Sci. Rep. 7, 8466 (2017)." href="/articles/s41929-023-01048-6#ref-CR44" id="ref-link-section-d10881139e3134">44</a></sup>. Through a PELE Global Exploration, we obtained an energy landscape profile. Further, in PELE Local Exploration, we analysed the energy landscape profile with respect to the catalytic distance between substrate ester reactive atoms and the nucleophile oxygen of the catalytic serine. Moreover, we estimated the relative activity of the active site for each substrate, calculating the number of catalytic events (equation (<a data-track="click" data-track-label="link" data-track-action="equation anchor" href="/articles/s41929-023-01048-6#Equ1">1</a>)). The number of catalytic events for each substrate was determined by multiple substrate-active site distance metrics. The FraC<sub>m1</sub> contains eight possible combinations of serine–histidine interactions between α-helices and two equidistant acids that can complete the catalytic triad, increasing the possible combinations. To avoid underestimation of catalytic events, we generated a distance matrix for each PELE step for all active sites and selected the catalytic triad nearest the substrate. For FraC<sub>m2,</sub> we focused on only one monomer globular part. For extensive details, see <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">Supplementary Methods</a>.</p><div id="Equ1" class="c-article-equation"><div class="c-article-equation__content"><span class="mathjax-tex">$${\mathrm{NRA}}=\,\frac{{\mathrm{number}}\,{\mathrm{of}}\,{\mathrm{catalytic}}\,{\mathrm{PELE}}\,{\mathrm{poses}}}{{\mathrm{total}}\,{\mathrm{number}}\,{\mathrm{of}}\,{\mathrm{accepted}}\,{\mathrm{PELE}}\,{\mathrm{steps}}\,\times\, {\mathrm{number}}\,{\mathrm{of}}\,{\mathrm{ester}}\,{\mathrm{groups}}} \%$$</span></div><div class="c-article-equation__number"> (1) </div></div><h3 class="c-article__sub-heading" id="Sec12">MD simulations of FraC<sub>m1</sub>, FraC<sub>m2</sub> and the polymer</h3><p>To analyse the stability of the membrane-inserted N-terminal domains in FraC<sub>m1/m2</sub>, MD simulations were performed using the Bilayer Membrane Builder protocol with the replacement method provided by CHARMM-GUI<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 45" title="Lee, J. et al. CHARMM-GUI input generator for NAMD, GROMACS, AMBER, OpenMM, and CHARMM/OpenMM simulations using the CHARMM36 additive force field. J. Chem. Theory Comput. 12, 405–413 (2016)." href="/articles/s41929-023-01048-6#ref-CR45" id="ref-link-section-d10881139e3280">45</a></sup> and GROMACS 5.1.2 simulations<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 46" title="Bauer, P., Hess, B., & Lindahl, E. GROMACS 2022 Manual. Zenodo https://doi.org/10.5281/ZENODO.6103568 " href="/articles/s41929-023-01048-6#ref-CR46" id="ref-link-section-d10881139e3284">46</a></sup>. To analyse the MD of the polymer, including its flexibility in the solvent of a 200-monomer PET fragment, MD simulations were performed using the Polymer Builder protocol provided by CHARMM-GUI<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 45" title="Lee, J. et al. CHARMM-GUI input generator for NAMD, GROMACS, AMBER, OpenMM, and CHARMM/OpenMM simulations using the CHARMM36 additive force field. J. Chem. Theory Comput. 12, 405–413 (2016)." href="/articles/s41929-023-01048-6#ref-CR45" id="ref-link-section-d10881139e3288">45</a></sup> and GROMACS 5.1.2 (ref. <sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 46" title="Bauer, P., Hess, B., & Lindahl, E. GROMACS 2022 Manual. Zenodo https://doi.org/10.5281/ZENODO.6103568 " href="/articles/s41929-023-01048-6#ref-CR46" id="ref-link-section-d10881139e3292">46</a></sup>). To obtain the different metrics of all MD simulations, the MDTraj<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 47" title="McGibbon, R. T. et al. MDTraj: a modern open library for the analysis of molecular dynamics trajectories. Biophys. J. 109, 1528–1532 (2015)." href="/articles/s41929-023-01048-6#ref-CR47" id="ref-link-section-d10881139e3297">47</a></sup> and MDAnalysis<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 48" title="Michaud-Agrawal, N., Denning, E. J., Woolf, T. B. & Beckstein, O. MDAnalysis: a toolkit for the analysis of molecular dynamics simulations. J. Comput. Chem. 32, 2319–2327 (2011)." href="/articles/s41929-023-01048-6#ref-CR48" id="ref-link-section-d10881139e3301">48</a></sup> Python modules were employed. For the simulation of the polymer, we used GROMACS<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 49" title="Abraham, M. J. et al. GROMACS: high performance molecular simulations through multi-level parallelism from laptops to supercomputers. SoftwareX 1–2, 19–25 (2015)." href="/articles/s41929-023-01048-6#ref-CR49" id="ref-link-section-d10881139e3305">49</a></sup> analysis tools to obtain its solvent-accessible surface area. For extensive details, see <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">Supplementary Methods</a>.</p><h3 class="c-article__sub-heading" id="Sec13">QM/MM minimizations</h3><p>QM/MM minimizations of the reactant state in <i>Is</i>PETase, FraC<sub>m1</sub> and FraC<sub>m2</sub> mutants were performed using Qsite from the Schrödinger Suite<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 50" title="Murphy, R. B., Philipp, D. M. & Friesner, R. A. A mixed quantum mechanics/molecular mechanics (QM/MM) method for large-scale modeling of chemistry in protein environments. J. Comput. Chem. 21, 1442–1457 (2000)." href="/articles/s41929-023-01048-6#ref-CR50" id="ref-link-section-d10881139e3327">50</a></sup>. The calculations were performed at the DFT/B3LYP and 6-31G* basis set level of theory. The QM region included <b>ETE</b> as a substrate and catalytic serine and histidine residues: Ser131 and His208 for <i>Is</i>PETase, Ser21 and His20 for FraC<sub>m1</sub> and Ser38 and His175 for FraC<sub>m2</sub>. The <i>Is</i>PETase reactant state’s structure was based on the PDB code <a href="https://www.rcsb.org/structure/5XH3">5XH3</a> (ref. <sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 51" title="5XH3: crystal structure of a novel PET hydrolase R103G/S131A mutant in complex with HEMT from Ideonella sakaiensis 201-F6. NCBI https://www.ncbi.nlm.nih.gov/Structure/pdb/5XH3 " href="/articles/s41929-023-01048-6#ref-CR51" id="ref-link-section-d10881139e3352">51</a></sup>), and both FraC mutant structures were based on a catalytic PELE pose.</p><h3 class="c-article__sub-heading" id="Sec14">npFraC<sub>WT</sub>, npFraC<sub>m1</sub> and npFraC<sub>m2</sub> production and assembly</h3><p>The complementary DNA encoding FraC<sub>m1</sub>, with Lys20His/Thr21Ser mutations, and FraC<sub>m2</sub>, with Asp38Ser/Glu173Gln mutations, were obtained via the overlap extension mutagenesis method<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 22" title="Alegre-Cebollada, J. et al. Silent mutations at the 5′-end of the cDNA of actinoporins from the sea anemone Stichodactyla helianthus allow their heterologous overproduction in Escherichia coli. J. Biotechnol. 127, 211–221 (2007)." href="/articles/s41929-023-01048-6#ref-CR22" id="ref-link-section-d10881139e3376">22</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 52" title="Sambrook, J. & Russell, D. W. In vitro mutagenesis using double-stranded DNA templates: selection of mutants with DpnI. Cold Spring Harb. Protoc. https://doi.org/10.1101/pdb.prot097766 (2018)." href="/articles/s41929-023-01048-6#ref-CR52" id="ref-link-section-d10881139e3379">52</a></sup>. FraC<sub>WT</sub>, FraC<sub>m1</sub> and FraC<sub>m2</sub> were produced in an <i>E. coli</i> expression system and purified as previously described<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 21" title="Tanaka, K., Caaveiro, J. M. M., Morante, K., González-Mañas, J. M. & Tsumoto, K. Structural basis for self-assembly of a cytolytic pore lined by protein and lipid. Nat. Commun. 6, 1–11 (2015)." href="/articles/s41929-023-01048-6#ref-CR21" id="ref-link-section-d10881139e3393">21</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 22" title="Alegre-Cebollada, J. et al. Silent mutations at the 5′-end of the cDNA of actinoporins from the sea anemone Stichodactyla helianthus allow their heterologous overproduction in Escherichia coli. J. Biotechnol. 127, 211–221 (2007)." href="/articles/s41929-023-01048-6#ref-CR22" id="ref-link-section-d10881139e3396">22</a></sup>. Pore assembly was performed to mimic the actual situation encountered by actinoporins in nature, as detailed in <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">Supplementary Methods</a>.</p><h3 class="c-article__sub-heading" id="Sec15">LCC<sub>WT</sub>, LCC<sub>WCCG</sub> and <i>Is</i>PETase: source and purification</h3><p>The genes coding for the cutinase LCC in its wild-type form (LCC<sub>WT</sub>)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 18" title="Cui, Y. et al. Computational redesign of a PETase for plastic biodegradation under ambient condition by the GRAPE strategy. ACS Catal. 11, 1340–1350 (2021)." href="/articles/s41929-023-01048-6#ref-CR18" id="ref-link-section-d10881139e3421">18</a></sup>, as well as the WCCG variant (LCC<sub>WCCG</sub>)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 19" title="Barth, M. et al. Effect of hydrolysis products on the enzymatic degradation of polyethylene terephthalate nanoparticles by a polyester hydrolase from Thermobifida fusca. Biochem. Eng. J. 93, 222–228 (2015)." href="/articles/s41929-023-01048-6#ref-CR19" id="ref-link-section-d10881139e3427">19</a></sup>, were codon-optimized for <i>E. coli</i> K12 and synthesized by Biomatik in pET21a(+) (Novagen) between the restriction sites NdeI and SalI and were kindly donated by Pablo Pérez-García (Universität Hamburg, Germany). The construct containing the gene coding for <i>Is</i>PETase fused to a maltose-binding protein in the backbone of pMAL-p4x was kindly donated by Sebastian Weigert (University of Bayreuth, Germany)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 20" title="Vogel, K. et al. Enzymatic degradation of polyethylene terephthalate nanoplastics analyzed in real time by isothermal titration calorimetry. Sci. Total Environ. 773, 145111 (2021)." href="/articles/s41929-023-01048-6#ref-CR20" id="ref-link-section-d10881139e3438">20</a></sup>. The proteins were produced and purified as reported and stored at 1.5 mg ml<sup>−1</sup> in 40 mM HEPES buffer pH 7.0 until use.</p><h3 class="c-article__sub-heading" id="Sec16">Sub-micro- and nano-sized PET degradation tests: time course and kinetics</h3><p>For the nPET<sub>b</sub>, nPET<sub>c</sub>, nPET<sub>GFa</sub> and nPET<sub>GFc</sub> degradation tests, the following conditions were utilized, if otherwise not indicated: [npFraC<sub>m1/m2</sub>, <i>Is</i>PETase and LCC<sub>WT/WCCG</sub>], 1.5 μg ml<sup>−1</sup>; [nPET<sub>GFa/GFc/b/c</sub>], 1.1 mg ml<sup>−1</sup>; reaction volume, 1,000 μl; <i>T</i>, 40 °C; pH, 7.0 (20 mM HEPES buffer); reaction time, 48 h. For conventional Michaelis‒Menten kinetic analysis, reaction conditions are listed as follows: [npFraC<sub>m1</sub>], 15 µg ml<sup>−1</sup>; [npFraC<sub>m2</sub>], 30 µg ml<sup>−1</sup>; [LCC<sub>WT</sub>], 132 µg ml<sup>−1</sup>; [nPET<sub>b</sub>], 0.38–4.4 mg ml<sup>−1</sup>; reaction volume, 50 μl; reaction time, 30 min (for npFraC<sub>m1</sub> and npFraC<sub>m2</sub>) or 60 min (for LCC<sub>WT</sub>); <i>T</i>, 40 °C; pH, 7.0 (20 mM HEPES buffer); 1,000 rpm. For inverse Michaelis‒Menten kinetic analysis, the following reaction conditions were used: [npFraC<sub>m1</sub>], 0–73 µg g<sup>−1</sup> nPET<sub>b</sub>; [npFraC<sub>m2</sub>], 0–118 µg g<sup>−1</sup> nPET<sub>b</sub>; [LCC<sub>WT</sub>], 0–118 µg g<sup>−1</sup> nPET<sub>b</sub>; [nPET<sub>b</sub>], 1.1 g l<sup>−1</sup>; reaction volume, 50 μl; <i>T</i>, 40 °C; pH, 7.0; agitation, 1,000 rpm; time of reaction, 30 min (for npFraC<sub>m1</sub> and npFraC<sub>m2</sub>) or 60 min (for LCC<sub>WT</sub>). Evaluation of the degradation of nPET<sub>b</sub> particles of different sizes using npFraC<sub>m1/m2</sub> was performed using the following reaction conditions: [npFraC<sub>m1/m2</sub>], 1.5 μg ml<sup>−1</sup>; [nPET<sub>b</sub>], 1.1 mg ml<sup>−1</sup>; reaction volume, 100 μl; <i>T</i>, 40 °C; pH, 7.0 (20 mM HEPES buffer); reaction time, 48 h. All reactions were maintained in 2-ml safe-lock Eppendorf polypropylene tubes (ref. 0030 120.094; Eppendorf AG) in a thermoshaker (model Thermomixer comfort, Eppendorf AG). The reactions were stopped, at the indicated times, by diluting ten times with dimethyl sulfoxide (from Merck Life Science) and the degradation products were immediately analysed by HPLC (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">Supplementary Methods</a>). In all cases, datasets were collected with a Varian Star LC workstation 6.41 (Varian) and analysed using Excel 2019 and SigmaPlot 14.5. Quantifications of degradation products were performed by HPLC on the basis of calibrations with purified standards, whose identity was confirmed by high-resolution MS (see raw MS data in Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM4">14</a>). All reactions were performed in triplicate (<i>n</i> = 3) with the following control reactions set up using the same amount of material: (1) soluble FraC<sub>WT</sub>, FraC<sub>m1</sub> and FraC<sub>m2</sub>; (2) npFraC<sub>WT</sub>; (3) no protein; and (4) nPET<sub>b</sub>, nPET<sub>c</sub>, nPET<sub>GFa</sub> and nPET<sub>GFc</sub>, without protein added.</p><h3 class="c-article__sub-heading" id="Sec17">Ester hydrolysis assays</h3><p>The hydrolysis of glyceryl tripropionate (ref. W328618), vinyl acetate (ref. V150-3), phenyl acetate (ref. 108723), <b>ETE</b> (ref. 465151), from Merck Life Science, and <b>TE</b> (purified as detailed in <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">Supplementary Methods</a>) was assayed using a pH indicator assay at 40 °C and pH 8.0 by monitoring the absorbance at 550 nm (extinction coefficient (<i>ε</i>) of phenol red, 8,450 M<sup>−1</sup> cm<sup>−1</sup>), as reported<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 26" title="Alonso, S. et al. Genetically engineered proteins with two active sites for enhanced biocatalysis and synergistic chemo- and biocatalysis. Nat. Catal. 3, 319–328 (2020)." href="/articles/s41929-023-01048-6#ref-CR26" id="ref-link-section-d10881139e3606">26</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 27" title="Roda, S. et al. A Plurizyme with transaminase and hydrolase activity catalyzes cascade reactions. Angew. Chem. Int. Ed. Engl. 61, e202207344 (2022)." href="/articles/s41929-023-01048-6#ref-CR27" id="ref-link-section-d10881139e3609">27</a></sup>. The hydrolysis of the model esters <i>p</i>-NP acetate (ref. N-8130; Merck Life Science), propionate (Santa Cruz Biotechnology, ref. sc-256813) and butyrate (ref. N-9876; Merck Life Science) was assessed by monitoring the continuous production of 4-nitrophenol at 348 nm (pH-independent isosbestic point, <i>ε</i> = 4,147 M<sup>−1</sup> cm<sup>−1</sup>)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 26" title="Alonso, S. et al. Genetically engineered proteins with two active sites for enhanced biocatalysis and synergistic chemo- and biocatalysis. Nat. Catal. 3, 319–328 (2020)." href="/articles/s41929-023-01048-6#ref-CR26" id="ref-link-section-d10881139e3624">26</a></sup>. In all cases, a Synergy HT Multi-Mode Microplate Reader with Gen5 2.00 software (Biotek Instruments) was used. All reactions were performed in triplicate (<i>n</i> = 3) with control reactions and background signals considered, as reported<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 26" title="Alonso, S. et al. Genetically engineered proteins with two active sites for enhanced biocatalysis and synergistic chemo- and biocatalysis. Nat. Catal. 3, 319–328 (2020)." href="/articles/s41929-023-01048-6#ref-CR26" id="ref-link-section-d10881139e3631">26</a>,<a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 27" title="Roda, S. et al. A Plurizyme with transaminase and hydrolase activity catalyzes cascade reactions. Angew. Chem. Int. Ed. Engl. 61, e202207344 (2022)." href="/articles/s41929-023-01048-6#ref-CR27" id="ref-link-section-d10881139e3634">27</a></sup>. For extensive details, see <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">Supplementary Methods</a>.</p><h3 class="c-article__sub-heading" id="Sec18">Reporting summary</h3><p>Further information on research design is available in the <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM2">Nature Portfolio Reporting Summary</a> linked to this article.</p></div></div></section> </div> <div> <section data-title="Data availability"><div class="c-article-section" id="data-availability-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="data-availability">Data availability</h2><div class="c-article-section__content" id="data-availability-content"> <p>The authors declare that the main data supporting the findings of this study are available within the paper and related Supplementary Information, Supplementary Data and Source Data files. The molecular simulations, the MD simulations and the quantum mechanical minimizations have been deposited at Zenodo (<a href="https://zenodo.org/deposit/7755566">https://zenodo.org/deposit/7755566</a>)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 53" title="Robles-Martín, A. et al. Sub-micro and nano-sized polyethylene terephthalate deconstruction with engineered pore-forming protein nanopores. Zenodo 
 https://zenodo.org/deposit/7755566
 
 (2023)." href="/articles/s41929-023-01048-6#ref-CR53" id="ref-link-section-d10881139e3744">53</a></sup> under the identifier <a href="https://doi.org/10.5281/zenodo.7755566">https://doi.org/10.5281/zenodo.7755566</a>. To use the archive, download the file and extract its contents to a local directory using appropriate software. The directory contains separate folders for each type of simulation, along with input, output and README files. <a data-track="click" data-track-label="link" data-track-action="section anchor" href="/articles/s41929-023-01048-6#Sec21">Source data</a> are provided with this paper (the mass spectrometry files can be analysed with the software MassLynx V4.1). 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Sub-micro and nano-sized polyethylene terephthalate deconstruction with engineered pore-forming protein nanopores. <i>Zenodo</i> <a href="https://zenodo.org/deposit/7755566" data-track="click_references" data-track-action="external reference" data-track-value="external reference" data-track-label="https://zenodo.org/deposit/7755566">https://zenodo.org/deposit/7755566</a> (2023).</p></li></ol><p class="c-article-references__download u-hide-print"><a data-track="click" data-track-action="download citation references" data-track-label="link" rel="nofollow" href="https://citation-needed.springer.com/v2/references/10.1038/s41929-023-01048-6?format=refman&flavour=references">Download references<svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-download-medium"></use></svg></a></p></div></div></div></section></div><section data-title="Acknowledgements"><div class="c-article-section" id="Ack1-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Ack1">Acknowledgements</h2><div class="c-article-section__content" id="Ack1-content"><p>This study was conducted under the auspices of the FuturEnzyme Project funded by the European Union’s Horizon 2020 Research and Innovation Programme under the auspices of the FuturEnzyme Project (grant agreement no. 101000327) and the PlasticsFatE project (grant agreement no. 95921), and Horizon Europe Research and Innovation Programme under grant agreement no. GA101060625 (Nymphe project). We also acknowledge financial support under grants PID2020-112758RB-I00 (M.F.), PDC2021-121534-I00 (M.F.), TED2021-130544B-I00 (M.F.), PID2019-106370RB-I00 (V.G.) and PID2019-105838RB-C31 (F.J.P.) from the Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación (AEI) (Digital Object Identifier MCIN/AEI/10.13039/501100011033), Fondo Europeo de Desarrollo Regional (ERDF) A way of making Europe and the European Union NextGenerationEU/PRTR, UCM-Banco Santander Grants PR87/19-22556 and PR108/20-26896 and UnaEuropa (Unano) SF2106 (to A.M.P.). S.G.-L. was supported by a Real Colegio Complutense Postdoctoral Fellowship for Distinguished Junior Scholars. S.R. thanks the Spanish Ministry of Science and Innovation for a PhD fellowship (FPU19/00608). D.H.-M. thanks Complutense University of Madrid and Banco Santander for a PhD fellowship (CT82/20/CT83/20). A.R.-M. thanks the Spanish Ministry of Science and Innovation for a PhD fellowship (PRE2020-091825) and the project PID2019-106370RB-I00. We thank M. J. Vicente for the ESI–MS analysis, performed at the Servicio Interdepartamental de Investigación (SIDI) from the Autonomous University of Madrid, Spain.</p></div></div></section><section aria-labelledby="author-information" data-title="Author information"><div class="c-article-section" id="author-information-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="author-information">Author information</h2><div class="c-article-section__content" id="author-information-content"><span class="c-article-author-information__subtitle u-visually-hidden" id="author-notes">Author notes</span><ol class="c-article-author-information__list"><li class="c-article-author-information__item" id="na1"><p>These authors contributed equally: Ana Robles-Martín, Rafael Amigot-Sánchez, Laura Fernandez-Lopez.</p></li></ol><h3 class="c-article__sub-heading" id="affiliations">Authors and Affiliations</h3><ol class="c-article-author-affiliation__list"><li id="Aff1"><p class="c-article-author-affiliation__address">Barcelona Supercomputing Center, Barcelona, Spain</p><p class="c-article-author-affiliation__authors-list">Ana Robles-Martín, Sergi Roda & Víctor Guallar</p></li><li id="Aff2"><p class="c-article-author-affiliation__address">Universitat de Barcelona, Barcelona, Spain</p><p class="c-article-author-affiliation__authors-list">Ana Robles-Martín</p></li><li id="Aff3"><p class="c-article-author-affiliation__address">Departamento de Bioquímica y Biología Molecular, Facultades de Medicina, Biología y Ciencias Químicas, Universidad Complutense, Madrid, Spain</p><p class="c-article-author-affiliation__authors-list">Rafael Amigot-Sánchez, Diego Heras-Márquez, Álvaro Martínez-del-Pozo & Sara García-Linares</p></li><li id="Aff4"><p class="c-article-author-affiliation__address">Instituto de Catalisis y Petroleoquimica, Consejo Superior de Investigaciones Científicas, Madrid, Spain</p><p class="c-article-author-affiliation__authors-list">Laura Fernandez-Lopez, Jose L. Gonzalez-Alfonso, Víctor Alcolea-Rodriguez, David Almendral, Cristina Coscolín, Francisco J. Plou, Raquel Portela, Miguel A. Bañares & Manuel Ferrer</p></li><li id="Aff5"><p class="c-article-author-affiliation__address">Institució Catalana de Recerca i Estudis Avançats, Barcelona, Spain</p><p class="c-article-author-affiliation__authors-list">Víctor Guallar</p></li></ol><div class="u-js-hide u-hide-print" data-test="author-info"><span class="c-article__sub-heading">Authors</span><ol class="c-article-authors-search u-list-reset"><li id="auth-Ana-Robles_Mart_n-Aff1-Aff2"><span class="c-article-authors-search__title u-h3 js-search-name">Ana Robles-Martín</span><div class="c-article-authors-search__list"><div class="c-article-authors-search__item c-article-authors-search__list-item--left"><a href="/search?author=Ana%20Robles-Mart%C3%ADn" class="c-article-button" data-track="click" data-track-action="author link - publication" data-track-label="link" rel="nofollow">View author publications</a></div><div class="c-article-authors-search__item c-article-authors-search__list-item--right"><p class="search-in-title-js c-article-authors-search__text">You can also search for this author in <span class="c-article-identifiers"><a class="c-article-identifiers__item" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=search&term=Ana%20Robles-Mart%C3%ADn" data-track="click" data-track-action="author link - pubmed" data-track-label="link" rel="nofollow">PubMed</a><span class="u-hide"> </span><a class="c-article-identifiers__item" href="http://scholar.google.co.uk/scholar?as_q=&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_sauthors=%22Ana%20Robles-Mart%C3%ADn%22&as_publication=&as_ylo=&as_yhi=&as_allsubj=all&hl=en" data-track="click" data-track-action="author link - scholar" data-track-label="link" rel="nofollow">Google Scholar</a></span></p></div></div></li><li id="auth-Rafael-Amigot_S_nchez-Aff3"><span class="c-article-authors-search__title u-h3 js-search-name">Rafael Amigot-Sánchez</span><div class="c-article-authors-search__list"><div class="c-article-authors-search__item c-article-authors-search__list-item--left"><a href="/search?author=Rafael%20Amigot-S%C3%A1nchez" class="c-article-button" data-track="click" data-track-action="author link - publication" data-track-label="link" rel="nofollow">View author publications</a></div><div class="c-article-authors-search__item c-article-authors-search__list-item--right"><p class="search-in-title-js c-article-authors-search__text">You can also search for this author in <span class="c-article-identifiers"><a class="c-article-identifiers__item" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=search&term=Rafael%20Amigot-S%C3%A1nchez" data-track="click" data-track-action="author link - pubmed" data-track-label="link" rel="nofollow">PubMed</a><span class="u-hide"> </span><a class="c-article-identifiers__item" href="http://scholar.google.co.uk/scholar?as_q=&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_sauthors=%22Rafael%20Amigot-S%C3%A1nchez%22&as_publication=&as_ylo=&as_yhi=&as_allsubj=all&hl=en" data-track="click" data-track-action="author link - scholar" data-track-label="link" rel="nofollow">Google Scholar</a></span></p></div></div></li><li id="auth-Laura-Fernandez_Lopez-Aff4"><span class="c-article-authors-search__title u-h3 js-search-name">Laura Fernandez-Lopez</span><div class="c-article-authors-search__list"><div class="c-article-authors-search__item c-article-authors-search__list-item--left"><a href="/search?author=Laura%20Fernandez-Lopez" class="c-article-button" data-track="click" data-track-action="author link - publication" data-track-label="link" rel="nofollow">View author publications</a></div><div class="c-article-authors-search__item c-article-authors-search__list-item--right"><p class="search-in-title-js c-article-authors-search__text">You can also search for this author in <span class="c-article-identifiers"><a class="c-article-identifiers__item" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=search&term=Laura%20Fernandez-Lopez" data-track="click" data-track-action="author link - pubmed" data-track-label="link" rel="nofollow">PubMed</a><span class="u-hide"> </span><a class="c-article-identifiers__item" href="http://scholar.google.co.uk/scholar?as_q=&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_sauthors=%22Laura%20Fernandez-Lopez%22&as_publication=&as_ylo=&as_yhi=&as_allsubj=all&hl=en" data-track="click" data-track-action="author link - scholar" data-track-label="link" rel="nofollow">Google Scholar</a></span></p></div></div></li><li id="auth-Jose_L_-Gonzalez_Alfonso-Aff4"><span class="c-article-authors-search__title u-h3 js-search-name">Jose L. 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pubmed" data-track-label="link" rel="nofollow">PubMed</a><span class="u-hide"> </span><a class="c-article-identifiers__item" href="http://scholar.google.co.uk/scholar?as_q=&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_sauthors=%22V%C3%ADctor%20Guallar%22&as_publication=&as_ylo=&as_yhi=&as_allsubj=all&hl=en" data-track="click" data-track-action="author link - scholar" data-track-label="link" rel="nofollow">Google Scholar</a></span></p></div></div></li></ol></div><h3 class="c-article__sub-heading" id="contributions">Contributions</h3><p>A.R.-M., R.A.-S., and L.F.-L. contributed equally to this work. V.G., S.G.-L. and M.F. lead this contribution. V.G., A.M.-P. and M.F. wrote the original manuscript, which was further prepared through the contributions of A.R.-M., S.R., S.G.-L., R.P. and M.A.B. All the authors have given approval to the final version of the manuscript. R.A.-S., D.H.-M., A.M.-P. and S.G.-L. contributed to the production, purification and haemolytic and structural characterization of the FraC pore-forming proteins. J.L.G.-A. and F.J.P. contributed to the HPLC analysis. L.F.-L., C.C., D.A. and M.F. performed the ester and nPET hydrolysis assays and determinations. A.R.-M., S.R. and V.G. contributed to the computational analysis. V.A.-R., R.P. and M.A.B. contributed to the FE-SEM analyses. V.G. originally conceived the computational analysis, which was further experimentally conceived by A.M.-P., S.G.-L. and M.F.</p><h3 class="c-article__sub-heading" id="corresponding-author">Corresponding authors</h3><p id="corresponding-author-list">Correspondence to <a id="corresp-c1" href="mailto:sglinares@ucm.es">Sara García-Linares</a>, <a id="corresp-c2" href="mailto:mferrer@icp.csic.es">Manuel Ferrer</a> or <a id="corresp-c3" href="mailto:victor.guallar@bsc.es">Víctor Guallar</a>.</p></div></div></section><section data-title="Ethics declarations"><div class="c-article-section" id="ethics-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="ethics">Ethics declarations</h2><div class="c-article-section__content" id="ethics-content"> <h3 class="c-article__sub-heading" id="FPar4">Competing interests</h3> <p>The authors declare no competing interests.</p> </div></div></section><section data-title="Peer review"><div class="c-article-section" id="peer-review-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="peer-review">Peer review</h2><div class="c-article-section__content" id="peer-review-content"> <h3 class="c-article__sub-heading" id="FPar3">Peer review information</h3> <p><i>Nature Catalysis</i> thanks Pedro Alexandrino Fernandes, Bian Wu, Ren Wei and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.</p> </div></div></section><section data-title="Additional information"><div class="c-article-section" id="additional-information-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="additional-information">Additional information</h2><div class="c-article-section__content" id="additional-information-content"><p><b>Publisher’s note</b> Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.</p></div></div></section><section data-title="Extended data"><div class="c-article-section" id="Sec19-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec19">Extended data</h2><div class="c-article-section__content" id="Sec19-content"><div data-test="supplementary-info"><div id="figshareContainer" class="c-article-figshare-container" data-test="figshare-container"></div><div class="c-article-supplementary__item js-c-reading-companion-figures-item" data-test="supp-item" id="Fig8"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="extended data fig. 1 chemical structures of the de" href="/articles/s41929-023-01048-6/figures/8" data-supp-info-image="//media.springernature.com/lw685/springer-static/esm/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig8_ESM.jpg">Extended Data Fig. 1 Chemical structures of the degradation products identified in the present study.</a></h3><div class="c-article-supplementary__description" data-component="thumbnail-container"><p>The figure was constructed using ChemDraw 18.2. <b>E</b> refers to ethylene glycol, <b>T</b> refers to terephthalic acid, <b>TE</b> refers to MHET, <b>ETE</b> refers to BHET, <b>TETE</b> refers to (MHET)<sub>2</sub> as reported by Joo et al. (2018)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 34" title="Joo, S. et al. Structural insight into molecular mechanism of poly(ethylene terephthalate) degradation. Nat. Commun. 9, 382 (2018)." href="/articles/s41929-023-01048-6#ref-CR34" id="ref-link-section-d10881139e3808">34</a></sup>, and <b>ETETE</b> refers to 2-HE(MHET)<sub>2</sub> as reported by Joo et al. (2018)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 34" title="Joo, S. et al. Structural insight into molecular mechanism of poly(ethylene terephthalate) degradation. Nat. Commun. 9, 382 (2018)." href="/articles/s41929-023-01048-6#ref-CR34" id="ref-link-section-d10881139e3817">34</a></sup>. Nomenclature as reported by Schubert et al.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 29" title="Schubert, S. et al. Reaction pathways for the enzymatic degradation of poly(ethylene terephthalate): What characterizes an efficient PET-hydrolase? ChemBioChem 24, e202200516 (2023)." href="/articles/s41929-023-01048-6#ref-CR29" id="ref-link-section-d10881139e3821">29</a></sup>.</p></div></div><div class="c-article-supplementary__item js-c-reading-companion-figures-item" data-test="supp-item" id="Fig9"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="extended data fig. 2 degradation profiles of npetg" href="/articles/s41929-023-01048-6/figures/9" data-supp-info-image="//media.springernature.com/lw685/springer-static/esm/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig9_ESM.jpg">Extended Data Fig. 2 Degradation profiles of nPET<sub>GFa/GFc/b/c</sub> treated with <i>Is</i>PETase and LCC<sub>WT/WCCG</sub>.</a></h3><div class="c-article-supplementary__description" data-component="thumbnail-container"><p><b>a-c</b> HPLC profiles of degradation products released under the experimental conditions: [<i>Is</i>PETase, LCC<sub>WT/WCCG</sub>], 1.5 μg ml<sup>-1</sup>; [nPET<sub>GFa/GFc/b/c</sub>], 1.1 mg ml<sup>-1</sup>; reaction volume, 1000 μl; T, 40 °C; pH, 7.0 (20 mM HEPES buffer); reaction time, 48 h. The reactions were maintained in 2-ml safe-lock Eppendorf® polypropylene tubes (ref. 0030 120.094) in a thermoshaker (model Thermomixer comfort, Eppendorf AG, Hamburg, Germany) at 1000 rpm. Aliquots of 100 μl were obtained, the reactions were stopped by adding 900 μl dimethyl sulfoxide (from Merck Life Science S.L.U., Madrid, Spain), and the degradation products were immediately analysed by HPLC. Datasets were collected with a Varian Star LC workstation 6.41 (Varian Inc., Palo Alto, California, USA). All reactions and analyses were performed in triplicate (<i>n</i> = 3), with a representative chromatogram per enzyme and nPET particle shown. <b>d-f</b> Concentration of degradation products from <b>T</b> to <b>ETETE</b> corresponding to a-e. Quantification was performed for products with unambiguous identification and for which enough material was recovered for HPLC calibration, namely, from <b>T</b> to <b>ETETE</b> (Extended Data Fig. <a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/s41929-023-01048-6#Fig8">1</a>); purification was performed by semipreparative HPLC, and the molecular weights and structures were determined by mass spectrometry (Supplementary Fig. <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM1">15</a>). Values are plotted as the mean of three independent replicates (<i>n</i> = 3) with the reported error ranges and SD calculated using the STDEV.S function in Excel 2019 (calculations and raw data are provided in Source Data). The figure was constructed using Excel 2019. Raw data are shown in Supplementary Data <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/s41929-023-01048-6#MOESM3">9</a>.</p></div></div><div class="c-article-supplementary__item js-c-reading-companion-figures-item" data-test="supp-item" id="Fig10"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="extended data fig. 3 schematic of the degradation " href="/articles/s41929-023-01048-6/figures/10" data-supp-info-image="//media.springernature.com/lw685/springer-static/esm/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_Fig10_ESM.jpg">Extended Data Fig. 3 Schematic of the degradation product profiles.</a></h3><div class="c-article-supplementary__description" data-component="thumbnail-container"><p>The degradation products identified after nPET hydrolysis by npFraC<sub>m1</sub>, npFraC<sub>m2</sub>, LCC<sub>WT</sub>, LCC<sub>WCCG</sub> and <i>Is</i>PETase are shown.</p></div></div><div class="c-article-supplementary__item" data-test="supp-item"><div class="c-article-table" data-test="inline-table" data-container-section="table" id="table-2"><figure><figcaption class="c-article-table__figcaption"><b id="Tab2" data-test="table-caption">Extended Data Table 1 Ester-hydrolysing activity of npFraC<sub>m1</sub> and npFraC<sub>m2</sub></b></figcaption><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="table-link" data-track="click" data-track-action="view table" data-track-label="button" rel="nofollow" href="/articles/s41929-023-01048-6/tables/2" aria-label="Full size table 2"><span>Full size table</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div></div><div class="c-article-supplementary__item" data-test="supp-item"><div class="c-article-table" data-test="inline-table" data-container-section="table" id="table-3"><figure><figcaption class="c-article-table__figcaption"><b id="Tab3" data-test="table-caption">Extended Data Table 2 Physical-chemical characteristics of the PET materials and nPET particles</b></figcaption><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="table-link" data-track="click" data-track-action="view table" data-track-label="button" rel="nofollow" href="/articles/s41929-023-01048-6/tables/3" aria-label="Full size table 3"><span>Full size table</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div></div></div></div></div></section><section data-title="Supplementary information"><div class="c-article-section" id="Sec20-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec20">Supplementary information</h2><div class="c-article-section__content" id="Sec20-content"><div data-test="supplementary-info"><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM1"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary information" href="https://static-content.springer.com/esm/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_MOESM1_ESM.pdf" data-supp-info-image="">Supplementary Information</a></h3><div class="c-article-supplementary__description" data-component="thumbnail-container"><p>Supplementary methods, Figs. 1–18, Notes 1–9 and references.</p></div></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM2"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="reporting summary" href="https://static-content.springer.com/esm/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_MOESM2_ESM.pdf" data-supp-info-image="">Reporting Summary</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM3"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary data 1–13" href="https://static-content.springer.com/esm/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_MOESM3_ESM.xlsx" data-supp-info-image="">Supplementary Data 1–13</a></h3><div class="c-article-supplementary__description" data-component="thumbnail-container"><p>Supplementary Data 1. Bioinformatic/computational raw data obtained from the different software simulations related to Figs. 1-4, Supplementary Figs. 2-6. Supplementary Data 2. Raw data for Supplementary Fig. 8 (Structural and functional comparison of FraC<sub>WT</sub> and FraC<sub>m1</sub> and FraC<sub>m2</sub>. Supplementary Data 3. Raw data (absorbance per min) and calculations for Extended Data Table 1 (Ester-hydrolysing activity of npFraC<sub>m1</sub> and npFraC<sub>m2</sub>). Supplementary Data 4. Raw data and calculations for Table 1 (Kinetic hydrolysis parameters of <b>ETE</b> and nPET<sub>b</sub> for npFraC<sub>m1</sub> and npFraC<sub>m2</sub>) and Supplementary Fig. 10 (Kinetic hydrolysis parameters of <b>ETE</b> for npFraC<sub>m1</sub> and npFraC<sub>m2</sub>). Supplementary Data 5. Raw data and calculations for Supplementary Fig. 11 (pH and thermal profiles of the npFraC<sub>m1</sub> and npFraC<sub>m2</sub>). Supplementary Data 6. Raw data for Supplementary Fig. 12 (Thermal denaturation of FraC<sub>WT</sub>, FraC<sub>m1</sub> and FraC<sub>m2</sub> as monitored by circular dichroism at 220 nm). Supplementary Data 7. Raw data and calculations for Fig. 5 (Particle size distribution of nPET<sub>GFa/GFc/b/c</sub> as determined by DLS). Supplementary Data 8. Raw data and calculations for Supplementary Fig. 14 (Thermal stability of npFraC<sub>m1</sub> and npFraC<sub>m2</sub> as monitored by activity measurements). Supplementary Data 9. Raw data and calculations for Fig. 6 (degradation profiles of nPET<sub>GFa/GFc/b/c</sub> treated with npFraC<sub>m1/m2</sub>) and Extended Data Fig. 2 (degradation profiles of nPET<sub>GFa/GFc/b/c</sub> treated with <i>Is</i>PETase and LCC<sub>WT/WCCG</sub>). Supplementary Data 10. Raw data and calculations for Fig. 7 (time course study of nPET<sub>b</sub> degradation using npFraC<sub>m1/m2</sub> compared with LCC<sub>WT</sub>). Supplementary Data 11. Raw data and calculations for Supplementary Fig. 16 (nPET<sub>c</sub> and PET<sub>GFa</sub> hydrolysis by LCC<sub>WT</sub> at 40 °C, 50 °C and 60 °C). Supplementary Data 12. Raw data and calculations for Supplementary Fig. 17a–c (conventional Michaelis–Menten (MM) kinetic study of nPET<sub>b</sub> particle degradation using npFraC<sub>m1/m2</sub> in comparison with the benchmark LCC<sub>WT</sub>) and the corresponding data in Table 1 (kinetic hydrolysis parameters of <b>ETE</b> and nPET<sub>b</sub> for npFraC<sub>m1</sub> and npFraC<sub>m2</sub>). Supplementary Data 13. Raw data and calculations for Supplementary Fig. 17d–f (inverse Michaelis–Menten (MM) kinetic study of nPET<sub>b</sub> particle degradation using npFraC<sub>m1/m2</sub> in comparison with the benchmark LCC<sub>WT</sub>) and the corresponding data in Table 1 (Kinetic hydrolysis parameters of <b>ETE</b> and nPET<sub>b</sub> for npFraC<sub>m1</sub> and npFraC<sub>m2</sub>).</p></div></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM4"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary data 14" href="https://static-content.springer.com/esm/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_MOESM4_ESM.zip" data-supp-info-image="">Supplementary Data 14</a></h3><div class="c-article-supplementary__description" data-component="thumbnail-container"><p>Raw MS data for Supplementary Fig. 15. The data in the .zip file can be analysed with the software MassLynx V4.1 (Waters) to obtain the mass spectra of the oligomers <b>ETE</b>, <b>TET</b>, <b>ETETE</b>, <b>TETE</b>, <b>TETET</b> and <b>TETETE</b>.</p></div></div></div></div></div></section><section data-title="Source data"><div class="c-article-section" id="Sec21-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec21">Source data</h2><div class="c-article-section__content" id="Sec21-content"><div data-test="supplementary-info"><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM5"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="source data supplementary fig. 7" href="https://static-content.springer.com/esm/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_MOESM5_ESM.jpg" data-supp-info-image="">Source Data Supplementary Fig. 7</a></h3><div class="c-article-supplementary__description" data-component="thumbnail-container"><p>Uncropped scan of the gel shown in Supplementary Fig. 7.</p></div></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM6"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="source data supplementary fig. 9" href="https://static-content.springer.com/esm/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_MOESM6_ESM.pdf" data-supp-info-image="">Source Data Supplementary Fig. 9</a></h3><div class="c-article-supplementary__description" data-component="thumbnail-container"><p>Original unprocessed negative stain electron microscopy micrographs, as shown in Supplementary Fig. 9. Original unprocessed negative stain electron microscopy micrograph of MSP1E3D1-assembled nanodiscs in the presence of FraC<sub>WT</sub>, FraC<sub>m1</sub> and FraC<sub>m2</sub>, in this order. Electron microscopy datasets have been collected with JEOL JEM1400 transmission electron microscope and processed using a DigitalMicrograph software (Gatan).</p></div></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM7"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="source data supplementary fig. 18" href="https://static-content.springer.com/esm/art%3A10.1038%2Fs41929-023-01048-6/MediaObjects/41929_2023_1048_MOESM7_ESM.pdf" data-supp-info-image="">Source Data Supplementary Fig. 18</a></h3><div class="c-article-supplementary__description" data-component="thumbnail-container"><p>Original unprocessed negative stain electron microscopy micrographs, as shown in Supplementary Fig. 18. In the order of appearance: non-treated nPET<sub>b</sub> (Supplementary Fig. 18a), treated nPET<sub>b</sub> with npFraC<sub>m1</sub> during 3 min, 15 min, 30 min and 24 h (Supplementary Fig. 18b), and treated nPET<sub>b</sub> with npFraC<sub>m2</sub> during 3 min, 15 min, 30 min and 24 h (Supplementary Fig. 18c). Magnification, 100,000×. Scale bar, 500 nm. FE-SEM was performed with an FEI Nova NanoSEM 230 microscope.</p></div></div></div></div></div></section><section data-title="Rights and permissions"><div class="c-article-section" id="rightslink-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="rightslink">Rights and permissions</h2><div class="c-article-section__content" id="rightslink-content"> <p><b>Open Access</b> This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit <a href="http://creativecommons.org/licenses/by/4.0/" rel="license">http://creativecommons.org/licenses/by/4.0/</a>.</p> <p class="c-article-rights"><a data-track="click" data-track-action="view rights and permissions" data-track-label="link" href="https://s100.copyright.com/AppDispatchServlet?title=Sub-micro-%20and%20nano-sized%20polyethylene%20terephthalate%20deconstruction%20with%20engineered%20protein%20nanopores&author=Ana%20Robles-Mart%C3%ADn%20et%20al&contentID=10.1038%2Fs41929-023-01048-6&copyright=The%20Author%28s%29&publication=2520-1158&publicationDate=2023-10-19&publisherName=SpringerNature&orderBeanReset=true&oa=CC%20BY">Reprints and permissions</a></p></div></div></section><section aria-labelledby="article-info" data-title="About this article"><div class="c-article-section" id="article-info-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="article-info">About this article</h2><div class="c-article-section__content" id="article-info-content"><div class="c-bibliographic-information"><div class="u-hide-print c-bibliographic-information__column c-bibliographic-information__column--border"><a data-crossmark="10.1038/s41929-023-01048-6" target="_blank" rel="noopener" href="https://crossmark.crossref.org/dialog/?doi=10.1038/s41929-023-01048-6" data-track="click" data-track-action="Click Crossmark" data-track-label="link" data-test="crossmark"><img loading="lazy" width="57" height="81" alt="Check for updates. 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