Concentration‐Dependent Inhibition of Mesophilic PETases on Poly(ethylene terephthalate) Can Be Eliminated by Enzyme Engineering

Avilan, Luisana; Lichtenstein, Bruce R.; König, Gerhard; Zahn, Michael; Allen, Mark D.; Oliveira, Liliana; Clark, Matilda; Bemmer, Victoria; Graham, Rosie; Austin, Harry P.; Dominick, Graham; Johnson, Christopher W.; Beckham, Gregg T.; McGeehan, John E.; Pickford, Andrew R.

doi:10.1002/cssc.202202277

Concentration‐Dependent Inhibition of Mesophilic PETases on Poly(ethylene terephthalate) Can Be Eliminated by Enzyme Engineering

Journal Article · Thu Mar 23 00:00:00 EDT 2023 · ChemSusChem

DOI:https://doi.org/10.1002/cssc.202202277· OSTI ID:1963133

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Centre for Enzyme Innovation School of Biological Sciences Institute of Biological and Biomedical Sciences University of Portsmouth Portsmouth PO1 2DY United Kingdom, BOTTLE Consortium Golden CO 80401 United States
Institute of Biochemistry Department of Biotechnology &, Enzyme Catalysis University of Greifswald D-17487 Greifswald Germany
Renewable Resources and Enabling Sciences Center National Renewable Energy Laboratory Golden CO 80401 United States
BOTTLE Consortium Golden CO 80401 United States, Renewable Resources and Enabling Sciences Center National Renewable Energy Laboratory Golden CO 80401 United States

Abstract

Enzyme‐based depolymerization is a viable approach for recycling of poly(ethylene terephthalate) (PET). PETase from Ideonella sakaiensis ( Is PETase) is capable of PET hydrolysis under mild conditions but suffers from concentration‐dependent inhibition. In this study, this inhibition is found to be dependent on incubation time, the solution conditions, and PET surface area. Furthermore, this inhibition is evident in other mesophilic PET‐degrading enzymes to varying degrees, independent of the level of PET depolymerization activity. The inhibition has no clear structural basis, but moderately thermostable Is PETase variants exhibit reduced inhibition, and the property is completely absent in the highly thermostable HotPETase, previously engineered by directed evolution, which simulations suggest results from reduced flexibility around the active site. This work highlights a limitation in applying natural mesophilic hydrolases for PET hydrolysis and reveals an unexpected positive outcome of engineering these enzymes for enhanced thermostability.

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE; USDOE Office of Energy Efficiency and Renewable Energy (EERE), Energy Efficiency Office. Advanced Materials & Manufacturing Office (AMMTO); USDOE Office of Energy Efficiency and Renewable Energy (EERE), Office of Sustainable Transportation. Bioenergy Technologies Office (BETO); University of Portsmouth

Grant/Contract Number:: AC36-08GO28308

OSTI ID:: 1963133

Alternate ID(s):: OSTI ID: 1962025
OSTI ID: 1971188

Report Number(s):: NREL/JA-2A00-84888; e202202277

Journal Information:: ChemSusChem, Journal Name: ChemSusChem Journal Issue: 8 Vol. 16; ISSN 1864-5631

Publisher:: Wiley Blackwell (John Wiley & Sons)Copyright Statement

Country of Publication:: Germany

Language:: English

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