Rational Protein Engineering to Increase the Activity and Stability of IsPETase Using the PROSS Algorithm

Andrew Philip Rennison, Jakob Winther*, Cristiano Varrone*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

13 Citations (Scopus)
63 Downloads (Pure)

Abstract

Polyethylene terephthalate (PET) is the most widely used polyester plastic, with applications in the textile and packaging industry. Currently, re-moulding is the main path for PET recycling, but this eventually leads to an unsustainable loss of quality; thus, other means of recycling are required. Enzymatic hydrolysis offers the possibility of monomer formation under mild conditions and opens up alternative and infinite recycling paths. Here, IsPETase, derived from the bacterium Ideonella sakaiensis, is considered to be the most active enzyme for PET degradation under mild conditions, and although several studies have demonstrated improvements to both the stability and activity of this enzyme, stability at even moderate temperatures is still an issue. In the present study, we have used sequence and structure-based bioinformatic tools to identify mutations to increase the thermal stability of the enzyme so as to increase PET degradation activity during extended hydrolysis reactions. We found that amino acid substitution S136E showed significant increases to activity and stability. S136E is a previously unreported variant that led to a 3.3-fold increase in activity relative to wild type.
Original languageEnglish
Article number3884
JournalPolymers
Volume13
Issue number22
Number of pages11
ISSN2073-4360
DOIs
Publication statusPublished - 10 Nov 2021

Keywords

  • PET hydrolysis
  • PETase
  • Thermal deactivation assay
  • Thermostability

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