PACER: a novel 3D plant cell wall model for the analysis of non-catalytic and enzymatic responses

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Volume Title

A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä

Date

2022-03-16

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en

Pages

11

Series

Biotechnology for Biofuels and Bioproducts, Volume 15, issue 1

Abstract

Background : Substrate accessibility remains a key limitation to the efficient enzymatic deconstruction of lignocellulosic biomass. Limited substrate accessibility is often addressed by increasing enzyme loading, which increases process and product costs. Alternatively, considerable efforts are underway world-wide to identify amorphogenesis-inducing proteins and protein domains that increase the accessibility of carbohydrate-active enzymes to targeted lignocellulose components.  Results : We established a three-dimensional assay, PACER (plant cell wall model for the analysis of non-catalytic and enzymatic responses), that enables analysis of enzyme migration through defined lignocellulose composites. A cellulose/azo-xylan composite was made to demonstrate the PACER concept and then used to test the migration and activity of multiple xylanolytic enzymes. In addition to non-catalytic domains of xylanases, the potential of loosenin-like proteins to boost xylanase migration through cellulose/azo-xylan composites was observed.  Conclusions : The PACER assay is inexpensive and parallelizable, suitable for screening proteins for ability to increase enzyme accessibility to lignocellulose substrates. Using the PACER assay, we visualized the impact of xylan-binding modules and loosenin-like proteins on xylanase mobility and access to targeted substrates. Given the flexibility to use different composite materials, the PACER assay presents a versatile platform to study impacts of lignocellulose components on enzyme access to targeted substrates.

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| openaire: EC/H2020/648925/EU//BHIVE Funding Information: We thank Kuisma Littunen and Kim Kataja who assisted with substrate preparation and Nelly Monties who assisted with enzyme production. We also thank Professor A. McGuigan (University of Toronto) for helpful discussions. Funding Information: We thank the following granting agencies for their financial support: the European Research Council (ERC) (Consolidator Grant no. BHIVE – 648925) (MM, EJ, VL and ERM); Jenny and Antti Wihuri Foundation (Centre for Young Synbio Scientists) (TK); Academy of Finland Postdoctoral Researcher Grant No. 331853 (TP); The French national research agency (anr) Grant No. ANR-18-CES43-DECO (TE and CD). Publisher Copyright: © 2022, The Author(s).

Keywords

Amorphogenesis, Assay development, Enzyme accessibility, Lignocellulose, Loosenin, Xylan, Xylanase

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Citation

Monschein, M, Jurak, E, Paasela, T, Koitto, T, Lambauer, V, Pavicic, M, Enjalbert, T, Dumon, C & Master, E R 2022, ' PACER: a novel 3D plant cell wall model for the analysis of non-catalytic and enzymatic responses ', Biotechnology for Biofuels and Bioproducts, vol. 15, no. 1, 30 . https://doi.org/10.1186/s13068-022-02128-8