A suspension-based assay and comparative detection methods for characterization of polyethylene terephthalate hydrolases

Jenny Arnling Bååth, Kim Borch, Peter Westh*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Enzymatic breakdown of plastic has emerged as a promising green technology, and its implementation will require assays that are accurate, reliable and convenient. Here, we assess two principles to monitor the hydrolysis of the common polyester, polyethylene terephthalate (PET). Hydrolysis of PET gives rise to heterogeneous products of different sizes and solubility, and as a result, specific experimental methods detect different activity levels. To avoid errors and to get a thorough picture of enzyme reactions, it is beneficial to combine several detection techniques. The two methods described herein are quantitative and complementary, and detect respectively the amount of soluble aromatic products and the formation of the constitutive aromatic monomers. A combined quantification approach identifies pitfalls in the characterization of these enzymes and provides mechanistic insight, but for screening and/or comparative studies of PET hydrolases we recommend a plate reader-based assay with suspended PET powder. This assay is rapid and simple, but still provides a good measure of the initial rates, which may be used in comparative biochemical analyses of these enzymes.
Original languageEnglish
Article number113873
JournalAnalytical Biochemistry
Volume607
Number of pages6
ISSN0003-2697
DOIs
Publication statusPublished - 2020

Keywords

  • PET hydrolase
  • Cutinase
  • Enzymatic assay
  • Product quantification
  • Spectrophotometric detection
  • RP-HPLC

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