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Abstract
Lignin is a complex heteroaromatic polymer which is one of the most abundant and diverse biopolymers on the planet. It comprises approximately one third of all woody plant matter, making it an attractive candidate as an alternative, renewable feedstock to petrochemicals to produce fine chemicals. However, the inherent complexity of lignin makes it difficult to analyse and characterise using common analytical techniques, proving a hindrance to the utilisation of lignin as a green chemical feedstock. Herein we outline the comprehensive tracking of lignin degradation by an alkaliphilic laccase in a semi-quantitative manner using a combined chemical analysis approach utilising Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) to characterise shifts in chemical diversity and relative abundance of ions, and NMR to highlight changes in the structure of lignin. Specifically, an alkaliphilic laccase was used to degrade an industrially relevant lignin, with compounds such as syringaresinol being almost wholly removed (95%) after 24 hours of treatment. Additionally, structural analyses reinforced these findings, indicating a >50% loss of signal relating to β-β linkages, of which syringaresinol is representative. Ultimately, this work underlines a combined analytical approach that can be used to gain a broader semi-quantitative understanding of the enzymatic activity of laccases within a complex, non-model mixture.
Supplementary materials
Title
Supplementary Figures and Tables
Description
Figure S1, Standard curve of different dilutions of Sigma Kraft Lignin (SKL).
Figure S2, The calculated coefficient of variation (CoV) for the quality control samples.
Table S1, Compounds used for spectra calibration of the digest samples and controls.
Table S2, Average Resolution and Assignment Error.
Table S3 ANOVA and Post-hoc Tukey Analysis Tables for Ion Count Stats
Table S4 Poisson GLM results between Time 0 and Time 2 and 24.
Table S5 Poisson GLM results between Time 2 and Time 0 and 24.
Table S6. Poisson GLM results between Time 0 and Time 2 and 24 for heteroatomic class of CHOx compounds.
Table S7. Poisson GLM results between Time 0 and Time 2 and 24 for heteroatomic class of CHOxS2 compounds.
Figure S3. van Krevelen Diagram of RI over the timecourse. Table S8, NMR Integration values for 24- hour laccase-treated and negative control samples.
Actions
Title
R Markdown File of Code for Analysis/Visualisation
Description
Code used for analysis & visualisation, contact first author for any feedback, questions and comments.
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Supplementary weblinks
Title
Horsfall Lab Datashare - Raw Data - Supplementary Figures and Tables
Description
Contains all raw NMR and FT-ICR MS spectra files, statistical outputs from R, and processed MS data.
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