Surface display as a functional screening platform for detecting enzymes active on PET

Sophia Andrea Helga Heyde, Jenny Arnling Bååth, Peter Westh, Morten H. H. Nørholm, Kenneth Jensen*

*Corresponding author for this work

Research output: Contribution to journalJournal articlepeer-review

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Abstract

Poly(ethylene terephthalate) (PET) is the world’s most abundant polyester plastic, and its ongoing accumulation in nature is causing a global environmental problem. Currently, the main recycling processes utilize thermomechanical or chemical means, resulting in the deterioration of the mechanical properties of PET. Consequently, polluting de novo synthesis remains preferred, creating the need for more efficient and bio-sustainable ways to hydrolyze the polymer. Recently, a PETase enzyme from the bacterium Ideonella sakaiensis was shown to facilitate PET biodegradation, albeit at slow rate. Engineering of more efficient PETases is required for industrial relevance, but progress is currently hampered by the dependency on intracellular expression in Escherichia coli. To create a more efficient screening platform in E. coli, we explore different surface display anchors for fast and easy assaying of PETase activity. We show that PETases can be functionally displayed on the bacterial cell surface, enabling screening of enzyme activity on PET microparticles – both while anchored to the cell and following solubilization of the enzymes.
Original languageEnglish
Article number93
JournalMicrobial Cell Factories
Volume20
Number of pages9
ISSN1475-2859
DOIs
Publication statusPublished - 2021

Keywords

  • PETase
  • Ideonella sakaiensis
  • Surface display
  • Extracellular protein production
  • E. coli

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