ACS Omega· 2026Q1
Development of a Vibrio natriegens -Based Plate-Clearing Assay for Rapid Screening of PET-Hydrolyzing Enzymes
- 2citations
- Q1SCImago
- 2026year
Short summary
A new plate-clearing assay using Vibrio natriegens enables rapid screening of PET-hydrolyzing enzymes, identifying a cutinase variant (T45P) with 3x higher PET-film degradation yield and favoring terephthalic acid production.
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Key points
- Developed a rapid plate-clearing assay using Vibrio natriegens for screening PET-hydrolyzing enzymes.
- Identified a cutinase variant (T45P) with 3-fold higher PET-film degradation yield than wild type.
- The T45P variant favored the production of terephthalic acid.
- The assay achieved a 25% hit rate when screening 150 colonies from an error-prone PCR library, demonstrating scalability.
AI-generated from the title and abstract; the full text is not read.
Abstract
Abstract Polyethylene terephthalate (PET) is a major contributor to plastic waste. Enzymatic PET degradation offers a sustainable recycling approach, but screening for effective enzymes remains challenging. Herein, we established an experimental plate-clearing assay using Vibrio natriegens, exploiting its protein secretion system to rapidly identify catalytic activity without time-consuming downstream processing. To validate the assay’s robustness, we tested variants of Fusarium solanipisi cutinase (FsC) and found that one variant, T45P, exhibited 3-fold higher PET-film degradation yield than the wild type and favored the production of terephthalic acid. As a further test case, we screened an error-prone polymerase chain reaction (PCR) library of FsC using the assay, obtaining a 25% hit rate after screening 150 colonies. This demonstrates the scalability of the method to large enzyme libraries.
The authors' abstract, as published at the source. ACS Omega, 2026 · DOI ↗
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BiotechnologyBiochemistry, Genetics and Molecular Biology