Systematic Assessment of Uncoupling in Flavoprotein Oxidases and Monooxygenases

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dc.contributor.authorGran-Scheuch, Alejandro
dc.contributor.authorParra, Loreto
dc.contributor.authorFraaije, Marco W.
dc.date.accessioned2025-01-20T20:15:12Z
dc.date.available2025-01-20T20:15:12Z
dc.date.issued2023
dc.description.abstractFlavin as a cofactor is an extremely versatile molecule that participates in a wide range of biochemical reactions. A special characteristic of the flavin cofactor, unique for a metal free cofactor, is its ability to react in its reduced form with molecular oxygen. This is exploited in flavoprotein oxidases and monooxygenases. The flavin-mediated reduction of dioxygen can lead to various reactive oxygen species (ROS), primarily hydrogen peroxide and superoxide. No systematic analysis of the formation of such reduced oxygen species produced by flavoprotein oxidases and monooxygenases had been performed before. In this work, we investigated which ROS are formed by several prototype flavoprotein oxidases and monooxygenases: phenylacetone monooxygenase (TfPAMO), an engineered TfPAMO variant that acts as an NADPH oxidase (TfPAMO C65D), eugenol oxidase (RjEUGO), and 5-hydroxymethyl furfural oxidase (MHMFO). The formed amounts of superoxide and hydrogen peroxide were measured under various conditions (different substrate concentrations, pH values and cosolvents). Interestingly, all flavoenzymes were found to produce, except for hydrogen peroxide, significant amounts of superoxide. Moreover, increased superoxide levels were measured at higher pH, which could be indicative for a pH-sensitive caged radical pair dissociation. To probe the effect of ROS formation on biocatalytic performance, conversions catalyzed by TfPAMO or RjEUGO, with or without catalase, were monitored. This revealed that catalase has a beneficial effect. No detrimental effect of the accumulation of superoxide on biocatalysis could be demonstrated. The results reveal that formation of ROS by flavoenzymes is highly dependent on the experimental conditions used. The results provide a better insight into the mechanism by which ROS is formed in flavoenzymes and may help studies or applications in which ROS formation should be promoted or minimized for industrial biocatalysis.
dc.fuente.origenWOS
dc.identifier.doi10.1021/acssuschemeng.1c02012
dc.identifier.issn2168-0485
dc.identifier.urihttps://doi.org/10.1021/acssuschemeng.1c02012
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/92246
dc.identifier.wosidWOS:000964208500001
dc.issue.numero13
dc.language.isoen
dc.pagina.final4959
dc.pagina.inicio4948
dc.revistaAcs sustainable chemistry & engineering
dc.rightsacceso restringido
dc.subjectflavoprotein monooxygenase
dc.subjectflavoprotein oxidase
dc.subjectflavin
dc.subjectreactive oxygen species (ROS)
dc.subjectsuperoxide
dc.subjecthydrogen peroxide
dc.subject.ods12 Responsible Consumption and Production
dc.subject.odspa12 Producción y consumo responsable
dc.titleSystematic Assessment of Uncoupling in Flavoprotein Oxidases and Monooxygenases
dc.typeartículo
dc.volumen11
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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