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

Mareike Monschein, Edita Jurak, Tanja Paasela, Taru Koitto, Vera Lambauer, Mirko Pavicic, Thomas Enjalbert, Claire Dumon, Emma R. Master*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

3 Citations (Scopus)
34 Downloads (Pure)

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.

Original languageEnglish
Article number30
Number of pages11
JournalBiotechnology for Biofuels and Bioproducts
Volume15
Issue number1
Early online date16 Mar 2022
DOIs
Publication statusPublished - 16 Mar 2022
MoE publication typeA1 Journal article-refereed

Keywords

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

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