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| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Recurso digital |
| Language: | |
| Published: |
Zenodo
2025
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| Online Access: | https://doi.org/10.5281/zenodo.17052114 |
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Table of 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>