A new continuous assay for quantitative assessment of enzymatic degradation of poly(ethylene terephthalate) (PET)

Thore Bach Thomsen, Sune W. Schubert, Cameron J. Hunt, Peter Westh, Anne S. Meyer*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

189 Downloads (Pure)


Enzymatic degradation of poly(ethylene terephthalate) (PET) has emerged as a promising route for ecofriendly biodegradation of plastic waste. Several discontinuous activity assays have been developed for assessing PET hydrolyzing enzymes, usually involving manual sampling at different time points during the course of the enzymatic reaction. In this work, we present a novel, compartmentalized UV absorbance assay for continuous detection of soluble hydrolysis products released during enzymatic degradation of PET. The methodology is based on removal of the walls separating two diagonally adjacent wells in UV-transparent microplates, to ensure passage of soluble enzymatic hydrolysis products between the two adjacent wells: One well holds an insoluble PET disk of defined dimensions and the other is used for continuous reading of the enzymatic product formation (at 240 nm). The assay was validated by quantifying the rate of mixing of the soluble PET degradation product BHET (bis(2-hydroxyethyl) terephthalate) between the two adjacent wells. The assay validation also involved a simple adjustment for water evaporation during prolonged assays. With this new assay, we determined the kinetic parameters for two PET hydrolases, DuraPETase and LCCICCG, and verified the underlying assumption of steady-state reaction rates. This new continuous assay enables fast exploration and robust kinetic characterization of PET degrading enzymes.
Original languageEnglish
Article number110142
JournalEnzyme and Microbial Technology
Number of pages7
Publication statusPublished - 2023


  • Enzymatic PET degradation
  • Assay development
  • PET hydrolase
  • Biotechnology
  • Interfacial enzymology
  • Heterogeneous catalysis


Dive into the research topics of 'A new continuous assay for quantitative assessment of enzymatic degradation of poly(ethylene terephthalate) (PET)'. Together they form a unique fingerprint.

Cite this