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Kinetics of H(2)O(2)-driven catalysis by a lytic polysaccharide monooxygenase from the fungus Trichoderma reesei
Owing to their ability to break glycosidic bonds in recalcitrant crystalline polysaccharides such as cellulose, the catalysis effected by lytic polysaccharide monooxygenases (LPMOs) is of major interest. Kinetics of these reductant-dependent, monocopper enzymes is complicated by the insoluble nature...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8528726/ https://www.ncbi.nlm.nih.gov/pubmed/34597668 http://dx.doi.org/10.1016/j.jbc.2021.101256 |
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author | Kuusk, Silja Väljamäe, Priit |
author_facet | Kuusk, Silja Väljamäe, Priit |
author_sort | Kuusk, Silja |
collection | PubMed |
description | Owing to their ability to break glycosidic bonds in recalcitrant crystalline polysaccharides such as cellulose, the catalysis effected by lytic polysaccharide monooxygenases (LPMOs) is of major interest. Kinetics of these reductant-dependent, monocopper enzymes is complicated by the insoluble nature of the cellulose substrate and parallel, enzyme-dependent, and enzyme-independent side reactions between the reductant and oxygen-containing cosubstrates. Here, we provide kinetic characterization of cellulose peroxygenase (oxidative cleavage of glycosidic bonds in cellulose) and reductant peroxidase (oxidation of the reductant) activities of the LPMO TrAA9A of the cellulose-degrading model fungus Trichoderma reesei. The catalytic efficiency [Formula: see text] of the cellulose peroxygenase reaction (k(cat) = 8.5 s(−1), and [Formula: see text]) was an order of magnitude higher than that of the reductant (ascorbic acid) peroxidase reaction. The turnover of H(2)O(2) in the ascorbic acid peroxidase reaction followed the ping-pong mechanism and led to irreversible inactivation of the enzyme with a probability of 0.0072. Using theoretical analysis, we suggest a relationship between the half-life of LPMO, the values of kinetic parameters, and the concentrations of the reactants. |
format | Online Article Text |
id | pubmed-8528726 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-85287262021-10-27 Kinetics of H(2)O(2)-driven catalysis by a lytic polysaccharide monooxygenase from the fungus Trichoderma reesei Kuusk, Silja Väljamäe, Priit J Biol Chem Research Article Owing to their ability to break glycosidic bonds in recalcitrant crystalline polysaccharides such as cellulose, the catalysis effected by lytic polysaccharide monooxygenases (LPMOs) is of major interest. Kinetics of these reductant-dependent, monocopper enzymes is complicated by the insoluble nature of the cellulose substrate and parallel, enzyme-dependent, and enzyme-independent side reactions between the reductant and oxygen-containing cosubstrates. Here, we provide kinetic characterization of cellulose peroxygenase (oxidative cleavage of glycosidic bonds in cellulose) and reductant peroxidase (oxidation of the reductant) activities of the LPMO TrAA9A of the cellulose-degrading model fungus Trichoderma reesei. The catalytic efficiency [Formula: see text] of the cellulose peroxygenase reaction (k(cat) = 8.5 s(−1), and [Formula: see text]) was an order of magnitude higher than that of the reductant (ascorbic acid) peroxidase reaction. The turnover of H(2)O(2) in the ascorbic acid peroxidase reaction followed the ping-pong mechanism and led to irreversible inactivation of the enzyme with a probability of 0.0072. Using theoretical analysis, we suggest a relationship between the half-life of LPMO, the values of kinetic parameters, and the concentrations of the reactants. American Society for Biochemistry and Molecular Biology 2021-09-28 /pmc/articles/PMC8528726/ /pubmed/34597668 http://dx.doi.org/10.1016/j.jbc.2021.101256 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Kuusk, Silja Väljamäe, Priit Kinetics of H(2)O(2)-driven catalysis by a lytic polysaccharide monooxygenase from the fungus Trichoderma reesei |
title | Kinetics of H(2)O(2)-driven catalysis by a lytic polysaccharide monooxygenase from the fungus Trichoderma reesei |
title_full | Kinetics of H(2)O(2)-driven catalysis by a lytic polysaccharide monooxygenase from the fungus Trichoderma reesei |
title_fullStr | Kinetics of H(2)O(2)-driven catalysis by a lytic polysaccharide monooxygenase from the fungus Trichoderma reesei |
title_full_unstemmed | Kinetics of H(2)O(2)-driven catalysis by a lytic polysaccharide monooxygenase from the fungus Trichoderma reesei |
title_short | Kinetics of H(2)O(2)-driven catalysis by a lytic polysaccharide monooxygenase from the fungus Trichoderma reesei |
title_sort | kinetics of h(2)o(2)-driven catalysis by a lytic polysaccharide monooxygenase from the fungus trichoderma reesei |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8528726/ https://www.ncbi.nlm.nih.gov/pubmed/34597668 http://dx.doi.org/10.1016/j.jbc.2021.101256 |
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