Development of a Vibrio natriegens -Based Plate-Clearing Assay for Rapid Screening of PET-Hydrolyzing Enzymes
- 1NTNU , , Sem Sælands vei 6/8 , ,
- 2Aalborg University , , ,
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.
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BibTeX
@article{trooyen2026development,
title = {{Development of a Vibrio natriegens -Based Plate-Clearing Assay for Rapid Screening of PET-Hydrolyzing Enzymes}},
author = {Susanne H. Troøyen and Ingrid M. Bolstad and Nora Drøivoldsmo and Davide Luciano and Evamaria I. Petersen and Gaston Courtade},
journal = {ACS Omega},
year = {2026},
month = nov,
doi = {10.1021/acsomega.6c05931},
url = {https://pubs.acs.org/acsodf/article/doi/10.1021/acsomega.6c05931/5464832/Development-of-a-Vibrio-natriegens-Based-Plate},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Development of a Vibrio natriegens -Based Plate-Clearing Assay for Rapid Screening of PET-Hydrolyzing Enzymes
AU - Susanne H. Troøyen
AU - Ingrid M. Bolstad
AU - Nora Drøivoldsmo
AU - Davide Luciano
AU - Evamaria I. Petersen
AU - Gaston Courtade
PY - 2026
DA - 2026/11/05
JO - ACS Omega
DO - 10.1021/acsomega.6c05931
UR - https://pubs.acs.org/acsodf/article/doi/10.1021/acsomega.6c05931/5464832/Development-of-a-Vibrio-natriegens-Based-Plate
AB - 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.
SN - 2470-1343
ER -