Concentration-dependent inhibition of mesophilic PETases on poly(ethylene terephthalate) can be eliminated by enzyme engineering

Luisana Avilan, Bruce Lichtenstein, Gerhard Koenig, Michael Zahn, Mark Devin Allen, Liliana Oliveira Pessoa, Matilda Alice Clark, Victoria Louise Bemmer, Rosie Graham, Harry P. Austin, Graham Dominick, Christopher W. Johnson, Gregg T. Beckham, John McGeehan, Andrew Pickford*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

7 Downloads (Pure)

Abstract

Enzyme-based depolymerization is a viable approach for recycling of poly(ethylene terephthalate) (PET). PETase from Ideonella sakaiensis (IsPETase) is capable of PET hydrolysis under mild conditions but suffers from concentration-dependent inhibition. Here, we report that this inhibition is dependent on incubation time, the solution conditions and PET surface area. Furthermore, this inhibition is evident in other mesophilic PET-degrading enzymes to varying degrees, independent of the level of PET depolymerization activity. The inhibition has no clear structural basis, but moderately thermostable IsPETase variants exhibit reduced inhibition, and the property is completely absent in the highly thermostable HotPETase, previously engineered by directed evolution, which our simulations suggest results from reduced flexibility around the active site. This work highlights a limitation in applying natural mesophilic hydrolases for PET hydrolysis, and reveals an unexpected positive outcome of engineering these enzymes for enhanced thermostability.
Original languageEnglish
JournalChemSusChem
Early online date21 Feb 2023
DOIs
Publication statusEarly online - 21 Feb 2023

Keywords

  • IsPETase
  • mesophilic enzymes
  • interfacial biocatalysis
  • inhibition
  • PET hydrolase

Cite this