Characterization of the auxiliary activity enzymes Trichoderma reesei TrAA3_2, TrAA9A and Podospora anserina PaAA9E, with potential roles in cellulose modification

Sustainably produced fuels and materials will be critical for the survival of our species, but a cost and time efficient production method is yet to be developed. Plant-based waste could be a part of the solution: one of its main components, cellulose, is a starting point for the production of biofu...

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Main Author: Hiltunen, Mimmu
Other Authors: Matemaattis-luonnontieteellinen tiedekunta, Faculty of Sciences, Bio- ja ympäristötieteiden laitos, Department of Biological and Environmental Science, Jyväskylän yliopisto, University of Jyväskylä
Format: Master's thesis
Language:eng
Published: 2021
Subjects:
Online Access: https://jyx.jyu.fi/handle/123456789/76324
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author Hiltunen, Mimmu
author2 Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä
author_facet Hiltunen, Mimmu Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä Hiltunen, Mimmu Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä
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description Sustainably produced fuels and materials will be critical for the survival of our species, but a cost and time efficient production method is yet to be developed. Plant-based waste could be a part of the solution: one of its main components, cellulose, is a starting point for the production of biofuels and chemicals. However, the recalcitrant nature of the main component of plant-based waste, lignocellulose, as well as the enigmatic nature of lignocellulolytic enzymes and their cooperation, has hindered adopting applications on a global scale. The aim of this study was to characterise three fungal enzymes with potential roles in lignocellulose degradation. First, we aimed to characterise the activity and phylogenetic location, as well as build a homology model, of a novel Trichoderma reesei AA3_2 subfamily enzyme (TrAA3_2). Secondly, we aimed to characterise the H2O2 dependency as well as thermal and pH stability of two lytic polysaccharide monooxygenases (TrAA9A and PaAA9E). Thirdly, we aimed to evaluate the effects of TrAA3_2, TrAA9A, and PaAA9E on the hydrolysis of spruce biomass by a cellulase mixture. We found that TrAA3_2 is phylogenetically closest to glucose oxidases but were unable to detect activity with any of the assayed carbohydrate or alcohol substrates. The homology model revealed that the substrate-binding residues of TrAA3_2 differ from those found in two well-characterised oxidative glucose active enzymes, possibly explaining the apparent inactivity for glucose. Furthermore, we found both TrAA9A and PaAA9E to be active with both O2 and H2O2 as a cosubstrate, and to be stable at 40 C and pH 5–7. TrAA9A was found to enhance the hydrolytic efficiency of a cellulase cocktail mimicking commercial lignocellulolytic cocktails. Based on our results, TrAA9A and PaAA9A display both oxidase and peroxidase activity, and may be good candidates for practical applications.
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spellingShingle Hiltunen, Mimmu Characterization of the auxiliary activity enzymes Trichoderma reesei TrAA3_2, TrAA9A and Podospora anserina PaAA9E, with potential roles in cellulose modification Solu- ja molekyylibiologia Cell and molecular biology 4013 Trichoderma reesei lignoselluloosa biomassa (teollisuus) hydrolyysi biopolttoaineet lignocellulose biomass hydrolysis biofuels
title Characterization of the auxiliary activity enzymes Trichoderma reesei TrAA3_2, TrAA9A and Podospora anserina PaAA9E, with potential roles in cellulose modification
title_full Characterization of the auxiliary activity enzymes Trichoderma reesei TrAA3_2, TrAA9A and Podospora anserina PaAA9E, with potential roles in cellulose modification
title_fullStr Characterization of the auxiliary activity enzymes Trichoderma reesei TrAA3_2, TrAA9A and Podospora anserina PaAA9E, with potential roles in cellulose modification Characterization of the auxiliary activity enzymes Trichoderma reesei TrAA3_2, TrAA9A and Podospora anserina PaAA9E, with potential roles in cellulose modification
title_full_unstemmed Characterization of the auxiliary activity enzymes Trichoderma reesei TrAA3_2, TrAA9A and Podospora anserina PaAA9E, with potential roles in cellulose modification Characterization of the auxiliary activity enzymes Trichoderma reesei TrAA3_2, TrAA9A and Podospora anserina PaAA9E, with potential roles in cellulose modification
title_short Characterization of the auxiliary activity enzymes Trichoderma reesei TrAA3_2, TrAA9A and Podospora anserina PaAA9E, with potential roles in cellulose modification
title_sort characterization of the auxiliary activity enzymes trichoderma reesei traa3 2 traa9a and podospora anserina paaa9e with potential roles in cellulose modification
title_txtP Characterization of the auxiliary activity enzymes Trichoderma reesei TrAA3_2, TrAA9A and Podospora anserina PaAA9E, with potential roles in cellulose modification
topic Solu- ja molekyylibiologia Cell and molecular biology 4013 Trichoderma reesei lignoselluloosa biomassa (teollisuus) hydrolyysi biopolttoaineet lignocellulose biomass hydrolysis biofuels
topic_facet 4013 Cell and molecular biology Solu- ja molekyylibiologia Trichoderma reesei biofuels biomass biomassa (teollisuus) biopolttoaineet hydrolysis hydrolyysi lignocellulose lignoselluloosa
url https://jyx.jyu.fi/handle/123456789/76324 http://www.urn.fi/URN:NBN:fi:jyu-202106083538
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