Mutational dissection of a hole hopping route in a lytic polysaccharide monooxygenase (LPMO).

Autor: Ayuso-Fernández I; Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), 1432, Ås, Norway. ivan.ayuso-fernandez@nmbu.no., Emrich-Mills TZ; Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), 1432, Ås, Norway., Haak J; Max Planck Institute for Chemical Energy Conversion, Stiftstrasse 34-36, 45470, Mülheim an der Ruhr, Germany.; Institute of Inorganic Chemistry, University of Duisburg-Essen, 45141, Essen, Germany., Golten O; Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), 1432, Ås, Norway., Hall KR; Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), 1432, Ås, Norway., Schwaiger L; Biocatalysis and Biosensing Laboratory, Department of Food Sciences and Technology, Institute of Food Science and Technology, University of Natural Resources and Life Sciences (BOKU), Muthgasse 18/2, Vienna, 1190, Austria., Moe TS; Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), 1432, Ås, Norway., Stepnov AA; Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), 1432, Ås, Norway., Ludwig R; Biocatalysis and Biosensing Laboratory, Department of Food Sciences and Technology, Institute of Food Science and Technology, University of Natural Resources and Life Sciences (BOKU), Muthgasse 18/2, Vienna, 1190, Austria., Cutsail Iii GE; Max Planck Institute for Chemical Energy Conversion, Stiftstrasse 34-36, 45470, Mülheim an der Ruhr, Germany.; Institute of Inorganic Chemistry, University of Duisburg-Essen, 45141, Essen, Germany., Sørlie M; Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), 1432, Ås, Norway., Kjendseth Røhr Å; Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), 1432, Ås, Norway., Eijsink VGH; Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), 1432, Ås, Norway. vincent.eijsink@nmbu.no.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2024 May 10; Vol. 15 (1), pp. 3975. Date of Electronic Publication: 2024 May 10.
DOI: 10.1038/s41467-024-48245-w
Abstrakt: Oxidoreductases have evolved tyrosine/tryptophan pathways that channel highly oxidizing holes away from the active site to avoid damage. Here we dissect such a pathway in a bacterial LPMO, member of a widespread family of C-H bond activating enzymes with outstanding industrial potential. We show that a strictly conserved tryptophan is critical for radical formation and hole transference and that holes traverse the protein to reach a tyrosine-histidine pair in the protein's surface. Real-time monitoring of radical formation reveals a clear correlation between the efficiency of hole transference and enzyme performance under oxidative stress. Residues involved in this pathway vary considerably between natural LPMOs, which could reflect adaptation to different ecological niches. Importantly, we show that enzyme activity is increased in a variant with slower radical transference, providing experimental evidence for a previously postulated trade-off between activity and redox robustness.
(© 2024. The Author(s).)
Databáze: MEDLINE
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