Redox robustness drives LPMO evolution

Enzymes known as lytic polysaccharide monooxygenases (LPMOs) are exceptionally powerful small redox enzymes that master the controlled generation and productive use of potentially damaging hydroxyl radicals in what is essentially a H<inf>2</inf>O<inf>2</inf>-driven peroxygena...

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Detalles Bibliográficos
Autores: Ayuso-Fernández, Iván, Emrich-Mills, Tom Z., Golten, Ole, Forsberg, Zarah, Hall, Kelsi R., Nagy, László G., Sørlie, Morten, Røhr, Åsmund Kjendseth, Eijsink, Vincent G. H.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2026
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/419896
Acceso en línea:http://hdl.handle.net/10261/419896
https://api.elsevier.com/content/abstract/scopus_id/105028578733
Access Level:acceso abierto
Palabra clave:Ancestral sequence reconstruction
Bacterial chitin oxidation
Lytic polysaccharide monooxygenases
Redox robustness evolution
Descripción
Sumario:Enzymes known as lytic polysaccharide monooxygenases (LPMOs) are exceptionally powerful small redox enzymes that master the controlled generation and productive use of potentially damaging hydroxyl radicals in what is essentially a H<inf>2</inf>O<inf>2</inf>-driven peroxygenase reaction. We have used ancestral sequence reconstruction and enzyme resurrection to unravel evolutionary steps leading to this exceptional catalytic ability. Real-time monitoring of copper reoxidation and amino acid radical formation showed evolutionary improvement of both the capacity to avoid futile turnover of H<inf>2</inf>O<inf>2</inf> and the ability to scavenge damaging radicals resulting from such turnover through a hole hopping pathway. Through mutational studies of ancestral LPMOs, we show that adoption of an extant-like conformation of residues in the hole hopping pathway yields improvements in redox robustness to near-extant levels. These results show how selective pressure imposed by the need for generating a highly oxidizing intermediate is a key driver of metalloenzyme evolution, involving large parts of the enzyme, well beyond the catalytic center.