Comparative biochemistry of PET hydrolase-carbohydrate-binding module fusion enzymes on a variety of PET substrates

Andrew Philip Rennison, Andreas Prestel, Peter Westh, Marie Sofie Møller*

*Corresponding author af dette arbejde

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

1 Downloads (Pure)

Abstract

Enzyme-driven recycling of PET has now become a fully developed industrial process. With the right pre-treatment, PET can be completely depolymerized within workable timeframes. This has been realized due to extensive research conducted over the past decade, resulting in a large set of engineered PET hydrolases. Among various engineering strategies to enhance PET hydrolases, fusion with binding domains has been used to tune affinity and boost activity of the enzymes. While fusion enzymes have demonstrated higher activity in many cases, these results are primarily observed under conditions that would not be economically viable at scale. Furthermore, the wide variation in PET substrates, conditions, and combinations of PET hydrolases and binding domains complicates direct comparisons. Here, we present a self-consistent and thorough analysis of two leading PET hydrolases, LCCICCG and PHL7. Both enzymes were evaluated both without and with a substrate-binding domain across a range of industrially relevant PET substrates. We demonstrate that the presence of a substrate-binding module does not significantly affect the affinity of LCCICCG and PHL7 for PET. However, significant differences exist in how the fusion enzymes act on different PET substrates and solid substrate loading, ranging from a 3-fold increase in activity to a 6-fold decrease. These findings could inform the tailoring of enzyme choice to different industrial scenarios.
OriginalsprogEngelsk
Artikelnummer110479
TidsskriftEnzyme and Microbial Technology
Vol/bind180
Antal sider13
ISSN0141-0229
DOI
StatusUdgivet - 2024

Bibliografisk note

Funding Information:
This work was supported by the Independent Research Fund Denmark [grant number: 1032-00273B]. NMR data were recorded at cOpenNMR, an infrastructure supported by the Novo Nordisk Foundation (NNF18OC0032996).

Publisher Copyright:
© 2024 The Authors

Citationsformater