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The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail

BACKGROUND: The discovery of enzymes named lytic polysaccharide monooxygenases (LPMOs) has had a major impact on the efficiency of current commercial cellulase cocktails for saccharification of lignocellulosic biomass. However, the notion that LPMOs use molecular oxygen as a co-substrate and require...

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Autores principales: Müller, Gerdt, Chylenski, Piotr, Bissaro, Bastien, Eijsink, Vincent G. H., Horn, Svein Jarle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6058378/
https://www.ncbi.nlm.nih.gov/pubmed/30061931
http://dx.doi.org/10.1186/s13068-018-1199-4
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author Müller, Gerdt
Chylenski, Piotr
Bissaro, Bastien
Eijsink, Vincent G. H.
Horn, Svein Jarle
author_facet Müller, Gerdt
Chylenski, Piotr
Bissaro, Bastien
Eijsink, Vincent G. H.
Horn, Svein Jarle
author_sort Müller, Gerdt
collection PubMed
description BACKGROUND: The discovery of enzymes named lytic polysaccharide monooxygenases (LPMOs) has had a major impact on the efficiency of current commercial cellulase cocktails for saccharification of lignocellulosic biomass. However, the notion that LPMOs use molecular oxygen as a co-substrate and require two externally delivered electrons per catalytic cycle poses a challenge in the development of efficient large-scale industrial processes. Building on the recent discovery that H(2)O(2), rather than O(2), is the co-substrate of LPMOs, we show here how cellulose degradation by the LPMO-containing commercial cellulase cocktail Cellic(®) CTec2 can be controlled and boosted by supplying the reaction with H(2)O(2). RESULTS: The controlled supply of anaerobic hydrolysis reactions with H(2)O(2) and sub-stoichiometric amounts of reductant increased apparent LPMO activity by almost two orders of magnitude compared to standard aerobic reactions utilizing O(2) and stoichiometric amounts of reductant. Improved LPMO activity was correlated with enhanced saccharification rates and yields for a model cellulosic substrate (Avicel) as well as industrial lignocellulosic substrates (sulfite-pulped Norway spruce and steam-exploded birch), although the magnitude of the effects was substrate dependent. Improvements in lignocellulose conversions were achieved at low H(2)O(2) feeding rates (in the range of 90–600 µM h(−1)). Tight control of LPMO reactions by controlled supply of H(2)O(2) under anaerobic conditions was possible. CONCLUSION: We report saccharification rates and yields for a model substrate (Avicel) and industrial lignocellulosic substrates that, at low H(2)O(2) feeding rates, are higher than those seen under standard aerobic conditions. In an industrial setting, controlling and supplying molecular oxygen and stoichiometric amounts of reductant are challenging. The present report shows that the use of small amounts of a liquid bulk chemical, H(2)O(2), provides an alternative to the currently available processes, which likely is cheaper and more easy to control, while giving higher product yields. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-018-1199-4) contains supplementary material, which is available to authorized users.
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spelling pubmed-60583782018-07-30 The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail Müller, Gerdt Chylenski, Piotr Bissaro, Bastien Eijsink, Vincent G. H. Horn, Svein Jarle Biotechnol Biofuels Research BACKGROUND: The discovery of enzymes named lytic polysaccharide monooxygenases (LPMOs) has had a major impact on the efficiency of current commercial cellulase cocktails for saccharification of lignocellulosic biomass. However, the notion that LPMOs use molecular oxygen as a co-substrate and require two externally delivered electrons per catalytic cycle poses a challenge in the development of efficient large-scale industrial processes. Building on the recent discovery that H(2)O(2), rather than O(2), is the co-substrate of LPMOs, we show here how cellulose degradation by the LPMO-containing commercial cellulase cocktail Cellic(®) CTec2 can be controlled and boosted by supplying the reaction with H(2)O(2). RESULTS: The controlled supply of anaerobic hydrolysis reactions with H(2)O(2) and sub-stoichiometric amounts of reductant increased apparent LPMO activity by almost two orders of magnitude compared to standard aerobic reactions utilizing O(2) and stoichiometric amounts of reductant. Improved LPMO activity was correlated with enhanced saccharification rates and yields for a model cellulosic substrate (Avicel) as well as industrial lignocellulosic substrates (sulfite-pulped Norway spruce and steam-exploded birch), although the magnitude of the effects was substrate dependent. Improvements in lignocellulose conversions were achieved at low H(2)O(2) feeding rates (in the range of 90–600 µM h(−1)). Tight control of LPMO reactions by controlled supply of H(2)O(2) under anaerobic conditions was possible. CONCLUSION: We report saccharification rates and yields for a model substrate (Avicel) and industrial lignocellulosic substrates that, at low H(2)O(2) feeding rates, are higher than those seen under standard aerobic conditions. In an industrial setting, controlling and supplying molecular oxygen and stoichiometric amounts of reductant are challenging. The present report shows that the use of small amounts of a liquid bulk chemical, H(2)O(2), provides an alternative to the currently available processes, which likely is cheaper and more easy to control, while giving higher product yields. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-018-1199-4) contains supplementary material, which is available to authorized users. BioMed Central 2018-07-24 /pmc/articles/PMC6058378/ /pubmed/30061931 http://dx.doi.org/10.1186/s13068-018-1199-4 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Müller, Gerdt
Chylenski, Piotr
Bissaro, Bastien
Eijsink, Vincent G. H.
Horn, Svein Jarle
The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail
title The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail
title_full The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail
title_fullStr The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail
title_full_unstemmed The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail
title_short The impact of hydrogen peroxide supply on LPMO activity and overall saccharification efficiency of a commercial cellulase cocktail
title_sort impact of hydrogen peroxide supply on lpmo activity and overall saccharification efficiency of a commercial cellulase cocktail
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6058378/
https://www.ncbi.nlm.nih.gov/pubmed/30061931
http://dx.doi.org/10.1186/s13068-018-1199-4
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