Enzyme Benchmarking with Polyethylene Furanoate Soluble Scaffolds for Directed Evolution of PEFases

Alcalde, Miguel

doi:10.1021/acsomega.4c09053

Published November 1, 2024 | Version v1

Journal article Open

Enzyme Benchmarking with Polyethylene Furanoate Soluble Scaffolds for Directed Evolution of PEFases

Alcalde, Miguel (Contact person)¹

1. Institute of Catalysis and Petrochemistry, ICP, CSIC

Contributors

Researcher (9):

1. EvoEnzyme S.L., Parque Científico de Madrid, Cantoblanco, 28049 Madrid, Spain
2. Department of Biocatalysis, Institute of Catalysis, CSIC, Marie Curie 2, Cantoblanco, 28049 Madrid, Spain. More by Dianelis T. Monterrey
3. Austrian Center of Industrial Biotechnology, Acib GmbH, Konrad-Lorenz-Straße 20, 3430 Tulln an der Donau, Austria
4. EvoEnzyme SL
5. EvoEnzyme S.L.
6. University of Genoa
7. Department of Agrobiotechnology IFA-Tulln, University of Natural Resources and Life Sciences Vienna, Institute of Environmental Biotechnology
8. Institute of Catalysis and Petrochemistry, ICP, CSIC

Plastic waste is a major threat in our industrialized world and is driving research into bioplastics. The success of biobased polyethylene furanoate (PEF) as a viable alternative to polyethylene terephthalate (PET) of fossil origin will depend on designing effective enzymes to break it down, aiding its recycling. Here, a panel of fungal and bacterial cutinases were functionally expressed in a tandem yeast expression system based on Saccharomyces cerevisiae and Pichia pastoris. The activity of the enzyme panel was tested with soluble PEF model scaffolds, observing a correlation with the degradation of real PEF powder. A high-throughput colorimetric screening assay based on the PEF scaffold diethyl furan-2,5-dicarboxylate was developed, establishing the basis for future directed evolution campaigns of PEFases.

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