De novo design of a polycarbonate hydrolase

Laura H. Holst, Niklas G. Madsen, Freja T. Toftgård, Freja Rønne, Ioana Malina Moise, Evamaria I. Petersen*, Peter Fojan*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

1 Citation (Scopus)

Abstract

Enzymatic degradation of plastics is currently limited to the use of engineered natural enzymes. As of yet, all engineering approaches applied to plastic degrading enzymes retain the natural α/β-fold. While mutations can be used to increase thermostability, an inherent maximum likely exists for the α/β-fold. It is thus of interest to introduce catalytic activity toward plastics in a different protein fold to escape the sequence space of plastic degrading enzymes. Here, a method for designing highly thermostable enzymes that can degrade plastics is described. With the help of Rosetta an active site catalysing the hydrolysis of polycarbonate is introduced into a set of thermostable scaffolds. Through computational evaluation, a potential PCase was selected and produced recombinantly in Escherichia coli. Thermal analysis suggests that the design has a melting temperature of >95C. Activity toward polycarbonate was confirmed using atomic force spectroscopy (AFM), proving the successful design of a PCase.

Original languageEnglish
Article numbergzad022
JournalProtein Engineering, Design and Selection
Volume36
ISSN1741-0126
DOIs
Publication statusPublished - 2023

Bibliographical note

Publisher Copyright:
© The Author(s) 2023. Published by Oxford University Press. All rights reserved.

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