A novel approach to analyze the impact of lytic polysaccharide monooxygenases (LPMOs) on cellulosic fibres

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Journal Title
Journal ISSN
Volume Title
A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä
Date
2024-03-15
Major/Subject
Mcode
Degree programme
Language
en
Pages
11
Series
Carbohydrate Polymers, Volume 328
Abstract
Enzymatic treatment of cellulosic fibres is a green alternative to classical chemical modification. For many applications, mild procedures for cellulose alteration are sufficient, in which the fibre structure and, therefore, the mechanical performance of cellulosic fibres are preserved. Lytic polysaccharide monooxygenases (LPMOs) bear a great potential to become a green reagent for such targeted cellulose modifications. An obstacle for wide implementation of LPMOs in tailored cellulose chemistry is the lack of suitable techniques to precisely monitor the LPMO impact on the polymer. Soluble oxidized cello-oligomers can be quantified using chromatographic and mass-spectrometric techniques. A considerable portion of the oxidized sites, however, remain on the insoluble cellulose fibres, and their quantification is difficult. Here, we describe a method for the simultaneous quantification of oxidized sites on cellulose fibres and changes in their molar mass distribution after treatment with LPMOs. The method is based on quantitative, heterogeneous, carbonyl-selective labelling with a fluorescent label (CCOA) followed by cellulose dissolution and size-exclusion chromatography (SEC). Application of the method to reactions of seven different LPMOs with pure cellulose fibres revealed pronounced functional differences between the enzymes, showing that this CCOA/SEC/MALS method is a promising tool to better understand the catalytic action of LPMOs.
Description
Publisher Copyright: © 2023 The Authors
Keywords
Carbonyl, Cellulose, Oxidation, Size exclusion chromatography, Solid fraction analysis
Other note
Citation
Sulaeva, I, Budischowsky, D, Rahikainen, J, Marjamaa, K, Støpamo, F G, Khaliliyan, H, Melikhov, I, Rosenau, T, Kruus, K, Várnai, A, Eijsink, V G H & Potthast, A 2024, ' A novel approach to analyze the impact of lytic polysaccharide monooxygenases (LPMOs) on cellulosic fibres ', Carbohydrate Polymers, vol. 328, 121696 . https://doi.org/10.1016/j.carbpol.2023.121696