Computationally guided genome rewiring of escherichia coli and its application for nanopolyethylene terephthalate (PET) biodegradation and upcycling

Numerous strategies for the biodegradation and upcycling of polyethylene terephthalate (PET) are under investigation. Here, we present a proof-of-concept study for reprogramming the Escherichia coli BL21(DE3) strain to degrade PET nanoparticles (nPET) without introducing foreign DNA and compromising native cellular fitness. In brief, native proteins selected in silico from the genome were repurposed to acquire artificial PETase activity without compromising their function and were subsequently replaced via CRISPR/Cas9 editing. A variant of the transport protein LsrB, selected for its ability to bind PET, was engineered to degrade PET powder (at 37–60°C). Building on LsrB periplasmic localization, we engineered a strain that degrades nPET at 37°C. The strain was further engineered to grow on nPET degradation products and produce valuable compounds. Our method, which is applicable across diverse genomes and microbial chassis, expands the potential of metabolic engineering to address plastic biodegradation and upcycling while reducing reliance on foreign DNA.

Keywords

CRISPR, Genome engineering, Metabolic engineering, Nanoplastics, Polyethylene terephthalate, Protein design, Protein engineering, Synthetic biology

Subject (DDC)

600 - Technik, Medizin, angewandte Wissenschaften

Project

Event

Exhibition start date

Exhibition end date

Conference start date

Conference end date

Date of the last check

ISBN

ISSN

1879-3096
0167-7799

Language

English

Created during FHNW affiliation

Yes

Strategic action fields FHNW

Publication status

Published

Review

Peer review of the complete publication

Open access category

Hybrid

License

Citation

Vidal Ramon, P., Shahgaldian, P., Gimenez Dejoz, J., Fernandez-Lopez, L., Romero, S., Nazemi, A., Luengo Perez, M., González Alfonso, J. L., Martínez Sugrañes, M., Robles Martín, A., Almendral Nieto, D., Roda, S., Pérez-García, P., Kruse, L., Jaeger, K.-E., Streit, W., Plou, F. J., Floor, M., Shahgaldian, P., et al. (2025). Computationally guided genome rewiring of escherichia coli and its application for nanopolyethylene terephthalate (PET) biodegradation and upcycling. Trends in Biotechnology, 43(11), 2874–2903. https://doi.org/10.1016/j.tibtech.2025.07.008

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