Functional screening pipeline to uncover laccase-like multicopper oxidase enzymes that transform industrial lignins

Anupama A. Sharan; Annie Bellemare; Marcos DiFalco; Adrian Tsang; Thu V. Vuong; Elizabeth A. Edwards; Emma R. Master

doi:10.1016/j.biortech.2023.130084

Functional screening pipeline to uncover laccase-like multicopper oxidase enzymes that transform industrial lignins

Anupama A. Sharan
, Annie Bellemare
, Marcos DiFalco
, Adrian Tsang
, Thu V. Vuong
, Elizabeth A. Edwards
, Emma R. Master^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Scientific › peer-review

8 Citations (Scopus)

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Abstract

Laccase-like multicopper oxidases are recognized for their potential to alter the reactivity of lignins for application in value-added products. Typically, model compounds are employed to discover such enzymes; however, they do not represent the complexity of industrial lignin substrates. In this work, a screening pipeline was developed to test enzymes simultaneously on model compounds and industrial lignins. A total of 12 lignin-active fungal multicopper oxidases were discovered, including 9 enzymes active under alkaline conditions (pH 11.0). Principal component analysis revealed the poor ability of model compounds to predict enzyme performance on industrial lignins. Additionally, sequence similarity analyses grouped these enzymes with Auxiliary Activity-1 sub-families with few previously characterized members, underscoring their taxonomic novelty. Correlation between the lignin-activity of these enzymes and their taxonomic origin, however, was not observed. These are critical insights to bridge the gap between enzyme discovery and application for industrial lignin valorization.

Original language	English
Article number	130084
Number of pages	10
Journal	Bioresource Technology
Volume	393
Early online date	1 Dec 2023
DOIs	https://doi.org/10.1016/j.biortech.2023.130084
Publication status	Published - Feb 2024
MoE publication type	A1 Journal article-refereed

Funding

The authors would like to acknowledge Mathieu Lavallee from Concordia University for support with heterologous production of multicopper oxidase enzymes presented in the study and Matthew Mascioni from the University of Toronto for development of R scripts used in ToF-SIMS data analysis. This work was supported by Genome Canada for the LSARP project “SYNBIOMICS – Functional genomics and techno-economic models for advanced biopolymer synthesis” (grant number 10405) and the NSERC CREATE for BioZone (grant number 528163). The authors would like to acknowledge Mathieu Lavallee from Concordia University for support with heterologous production of multicopper oxidase enzymes presented in the study and Matthew Mascioni from the University of Toronto for development of R scripts used in ToF-SIMS data analysis. This work was supported by Genome Canada for the LSARP project “SYNBIOMICS – Functional genomics and techno-economic models for advanced biopolymer synthesis” (grant number 10405) and the NSERC CREATE for BioZone (grant number 528163).

Keywords

Kraft lignin
Laccase
Lignin-active enzymes
Organosolv lignin
ToF-SIMS

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CHEM_Sharan_et_al_Functional_Screening_2023_Bioresource_TechnologyFinal published version, 3.37 MBLicence: CC BY

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title = "Functional screening pipeline to uncover laccase-like multicopper oxidase enzymes that transform industrial lignins",

abstract = "Laccase-like multicopper oxidases are recognized for their potential to alter the reactivity of lignins for application in value-added products. Typically, model compounds are employed to discover such enzymes; however, they do not represent the complexity of industrial lignin substrates. In this work, a screening pipeline was developed to test enzymes simultaneously on model compounds and industrial lignins. A total of 12 lignin-active fungal multicopper oxidases were discovered, including 9 enzymes active under alkaline conditions (pH 11.0). Principal component analysis revealed the poor ability of model compounds to predict enzyme performance on industrial lignins. Additionally, sequence similarity analyses grouped these enzymes with Auxiliary Activity-1 sub-families with few previously characterized members, underscoring their taxonomic novelty. Correlation between the lignin-activity of these enzymes and their taxonomic origin, however, was not observed. These are critical insights to bridge the gap between enzyme discovery and application for industrial lignin valorization.",

keywords = "Kraft lignin, Laccase, Lignin-active enzymes, Organosolv lignin, ToF-SIMS",

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note = "Funding Information: The authors would like to acknowledge Mathieu Lavallee from Concordia University for support with heterologous production of multicopper oxidase enzymes presented in the study and Matthew Mascioni from the University of Toronto for development of R scripts used in ToF-SIMS data analysis. This work was supported by Genome Canada for the LSARP project “SYNBIOMICS – Functional genomics and techno-economic models for advanced biopolymer synthesis” (grant number 10405) and the NSERC CREATE for BioZone (grant number 528163). Publisher Copyright: {\textcopyright} 2023",

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AU - Vuong, Thu V.

AU - Edwards, Elizabeth A.

AU - Master, Emma R.

N1 - Funding Information: The authors would like to acknowledge Mathieu Lavallee from Concordia University for support with heterologous production of multicopper oxidase enzymes presented in the study and Matthew Mascioni from the University of Toronto for development of R scripts used in ToF-SIMS data analysis. This work was supported by Genome Canada for the LSARP project “SYNBIOMICS – Functional genomics and techno-economic models for advanced biopolymer synthesis” (grant number 10405) and the NSERC CREATE for BioZone (grant number 528163). Publisher Copyright: © 2023

PY - 2024/2

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N2 - Laccase-like multicopper oxidases are recognized for their potential to alter the reactivity of lignins for application in value-added products. Typically, model compounds are employed to discover such enzymes; however, they do not represent the complexity of industrial lignin substrates. In this work, a screening pipeline was developed to test enzymes simultaneously on model compounds and industrial lignins. A total of 12 lignin-active fungal multicopper oxidases were discovered, including 9 enzymes active under alkaline conditions (pH 11.0). Principal component analysis revealed the poor ability of model compounds to predict enzyme performance on industrial lignins. Additionally, sequence similarity analyses grouped these enzymes with Auxiliary Activity-1 sub-families with few previously characterized members, underscoring their taxonomic novelty. Correlation between the lignin-activity of these enzymes and their taxonomic origin, however, was not observed. These are critical insights to bridge the gap between enzyme discovery and application for industrial lignin valorization.

AB - Laccase-like multicopper oxidases are recognized for their potential to alter the reactivity of lignins for application in value-added products. Typically, model compounds are employed to discover such enzymes; however, they do not represent the complexity of industrial lignin substrates. In this work, a screening pipeline was developed to test enzymes simultaneously on model compounds and industrial lignins. A total of 12 lignin-active fungal multicopper oxidases were discovered, including 9 enzymes active under alkaline conditions (pH 11.0). Principal component analysis revealed the poor ability of model compounds to predict enzyme performance on industrial lignins. Additionally, sequence similarity analyses grouped these enzymes with Auxiliary Activity-1 sub-families with few previously characterized members, underscoring their taxonomic novelty. Correlation between the lignin-activity of these enzymes and their taxonomic origin, however, was not observed. These are critical insights to bridge the gap between enzyme discovery and application for industrial lignin valorization.

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Functional screening pipeline to uncover laccase-like multicopper oxidase enzymes that transform industrial lignins

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Keywords

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