Fast and Specific Peroxygenase Reactions Catalyzed by Fungal Mono-Copper Enzymes
| Autor: | Anton A. Stepnov, Morten Sørlie, Lukas Rieder, Vincent G. H. Eijsink |
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| Rok vydání: | 2021 |
| Předmět: |
Biomass
chemistry.chemical_element Lentinula Polysaccharide Biochemistry Article Mixed Function Oxygenases Catalysis Fungal Proteins Polysaccharides Catalytic Domain Enzyme Assays chemistry.chemical_classification Neurospora crassa Hydrogen Peroxide Monooxygenase Copper Recombinant Proteins Kinetics Biodegradation Environmental Enzyme chemistry Lytic cycle Degradation (geology) |
| Zdroj: | Biochemistry |
| ISSN: | 1520-4995 0006-2960 |
| Popis: | The copper-dependent lytic polysaccharide monooxygenases (LPMOs) are receiving attention because of their role in the degradation of recalcitrant biomass and their intriguing catalytic properties. The fundamentals of LPMO catalysis remain somewhat enigmatic as the LPMO reaction is affected by a multitude of LPMO- and co-substrate-mediated (side) reactions that result in a complex reaction network. We have performed kinetic studies with two LPMOs that are active on soluble substrates, NcAA9C and LsAA9A, using various reductants typically employed for LPMO activation. Studies with NcAA9C under “monooxygenase” conditions showed that the impact of the reductant on catalytic activity is correlated with the hydrogen peroxide-generating ability of the LPMO-reductant combination, supporting the idea that a peroxygenase reaction is taking place. Indeed, the apparent monooxygenase reaction could be inhibited by a competing H2O2-consuming enzyme. Interestingly, these fungal AA9-type LPMOs were found to have higher oxidase activity than bacterial AA10-type LPMOs. Kinetic analysis of the peroxygenase activity of NcAA9C on cellopentaose revealed a fast stoichiometric conversion of high amounts of H2O2 to oxidized carbohydrate products. A kcat value of 124 ± 27 s–1 at 4 °C is 20 times higher than a previously described kcat for peroxygenase activity on an insoluble substrate (at 25 °C) and some 4 orders of magnitude higher than typical “monooxygenase” rates. Similar studies with LsAA9A revealed differences between the two enzymes but confirmed fast and specific peroxygenase activity. These results show that the catalytic site arrangement of LPMOs provides a unique scaffold for highly efficient copper redox catalysis. |
| Databáze: | OpenAIRE |
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