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Main Authors: Gimenez Dejoz, Joan, Vidal Ramon, Paula, Romero-Téllez, Sonia, Luengo Perez, Miguel, González Alfonso, José Luis, Martínez Sugrañes, Mireia, Robles Martín, Ana, Coscolín, Cristina, Roda, Sergi, Plou, Francisco J., Floor, Martin, Bargiela, Rafael, Ferrer, Manuel, Guallar, Victor, Fernandez-Lopez, Laura
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Published: Zenodo 2025
Online Access:https://doi.org/10.5281/zenodo.17052114
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author Gimenez Dejoz, Joan
Vidal Ramon, Paula
Romero-Téllez, Sonia
Luengo Perez, Miguel
González Alfonso, José Luis
Martínez Sugrañes, Mireia
Robles Martín, Ana
Coscolín, Cristina
Roda, Sergi
Plou, Francisco J.
Floor, Martin
Bargiela, Rafael
Ferrer, Manuel
Guallar, Victor
Fernandez-Lopez, Laura
author_facet Gimenez Dejoz, Joan
Vidal Ramon, Paula
Romero-Téllez, Sonia
Luengo Perez, Miguel
González Alfonso, José Luis
Martínez Sugrañes, Mireia
Robles Martín, Ana
Coscolín, Cristina
Roda, Sergi
Plou, Francisco J.
Floor, Martin
Bargiela, Rafael
Ferrer, Manuel
Guallar, Victor
Fernandez-Lopez, Laura
contents <p><span>The development of synthetic biology and metabolic engineering strategies offers a promising route to advance polyethylene terephthalate (PET) biodegradation and upcycling. </span><span>H</span><span>ere we optimized the computational GenRewire pipeline and successfully repurposed an endogenous outer membrane protein in Escherichia coli BL21 (DE3), enabling this bacterium</span><span>, </span><span>naturally unable to degrade PET</span><span>, </span><span>to depolymerize PET powder. This was achieved through a genome-editing strategy that combi</span><span>ned </span><span> catalytic triad design </span><span> with CRISPR–Cas9-based genomic replacement, resulting in a functional artificial PETase integrated directly into </span><span>the structure of</span><span> the outer membrane protein OmpC</span><span> and in </span><span>the</span><span> bacterium</span><span> genome. Under conditions of an initial optical density (</span><span>OD600</span><span> nm</span><span>) of 0.1 and a PET powder load of 30 mg/mL (<300 µm, >40% crystallinity), the engineered strain degraded PET as its sole carbon source at </span><span>37 °C, releasing up to 157 ± 2 µM of hydrolysis products after 24 h. These findings demonstrate the potential of genome rewiring to create fully endogenous, programmable microbial platforms for PET plastic biodegradation, without the need for exogenous DNA</span></p>
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id zenodo_https___doi_org_10_5281_zenodo_17052114
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publishDate 2025
publisher Zenodo
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spellingShingle Rewiring an Endogenous Outer Membrane Protein Enables Escherichia coli to Biodegrade PET Powder
Gimenez Dejoz, Joan
Vidal Ramon, Paula
Romero-Téllez, Sonia
Luengo Perez, Miguel
González Alfonso, José Luis
Martínez Sugrañes, Mireia
Robles Martín, Ana
Coscolín, Cristina
Roda, Sergi
Plou, Francisco J.
Floor, Martin
Bargiela, Rafael
Ferrer, Manuel
Guallar, Victor
Fernandez-Lopez, Laura
<p><span>The development of synthetic biology and metabolic engineering strategies offers a promising route to advance polyethylene terephthalate (PET) biodegradation and upcycling. </span><span>H</span><span>ere we optimized the computational GenRewire pipeline and successfully repurposed an endogenous outer membrane protein in Escherichia coli BL21 (DE3), enabling this bacterium</span><span>, </span><span>naturally unable to degrade PET</span><span>, </span><span>to depolymerize PET powder. This was achieved through a genome-editing strategy that combi</span><span>ned </span><span> catalytic triad design </span><span> with CRISPR–Cas9-based genomic replacement, resulting in a functional artificial PETase integrated directly into </span><span>the structure of</span><span> the outer membrane protein OmpC</span><span> and in </span><span>the</span><span> bacterium</span><span> genome. Under conditions of an initial optical density (</span><span>OD600</span><span> nm</span><span>) of 0.1 and a PET powder load of 30 mg/mL (<300 µm, >40% crystallinity), the engineered strain degraded PET as its sole carbon source at </span><span>37 °C, releasing up to 157 ± 2 µM of hydrolysis products after 24 h. These findings demonstrate the potential of genome rewiring to create fully endogenous, programmable microbial platforms for PET plastic biodegradation, without the need for exogenous DNA</span></p>
title Rewiring an Endogenous Outer Membrane Protein Enables Escherichia coli to Biodegrade PET Powder
url https://doi.org/10.5281/zenodo.17052114