H2-driven reduction of flavin by hydrogenase enables cleaner operation of nitroreductases for nitro-group to amine reductions
Hydrogenase-mediated reduction of flavin mononucleotide by H2 is exploited to enable cleaner application of nitroreductase enzymes for reduction of aromatic nitro functional groups. This turns the overall reaction into a biocatalytic hydrogenation. Use of flavin-containing nitroreductases in industr...
| Үндсэн зохиолчид: | , , , , , |
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| Формат: | Journal article |
| Хэл сонгох: | English |
| Хэвлэсэн: |
Frontiers Media
2022
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| _version_ | 1826308662139813888 |
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| author | Ramirez, MA Joseph Srinivasan, S Cleary, SE Todd, PMT Reeve, HA Vincent, KA |
| author_facet | Ramirez, MA Joseph Srinivasan, S Cleary, SE Todd, PMT Reeve, HA Vincent, KA |
| author_sort | Ramirez, MA |
| collection | OXFORD |
| description | Hydrogenase-mediated reduction of flavin mononucleotide by H2 is exploited to enable cleaner application of nitroreductase enzymes for reduction of aromatic nitro functional groups. This turns the overall reaction into a biocatalytic hydrogenation. Use of flavin-containing nitroreductases in industrial biotechnology typically relies upon NADH or NADPH as reductant, together with glucose dehydrogenase and glucose as a regeneration system for the reduced nicotinamide cofactor, with 3 equivalents of the carbon-intensive glucose required for a single 6-electron nitro to amine conversion. We show here that reduced flavin mononucleotide is an alternative reductant for nitroreductases, and by combining this with H2-driven recycling of reduced flavin, we avoid glucose, thereby enabling atom-efficient biocatalytic nitro reductions. We compare this biocatalytic system, via green chemistry metrics, to existing strategies for biocatalytic nitro-group reductions, particularly with respect to replacing glucose with H2 gas. We take steps towards demonstrating industrial viability: we report an overexpression system for E. coli hydrogenase 1, giving a 12-fold improvement in enzyme yield; we show a reaction in which the hydrogenase exhibits > 26,000 enzyme turnovers; and we demonstrate reasonable solvent tolerance of the hydrogenase and flavin reduction system which would enable reaction intensification. |
| first_indexed | 2024-03-07T07:22:40Z |
| format | Journal article |
| id | oxford-uuid:4ba5e449-aaac-4c84-9fdc-ca7df79ea294 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:22:40Z |
| publishDate | 2022 |
| publisher | Frontiers Media |
| record_format | dspace |
| spelling | oxford-uuid:4ba5e449-aaac-4c84-9fdc-ca7df79ea2942022-11-01T09:47:32ZH2-driven reduction of flavin by hydrogenase enables cleaner operation of nitroreductases for nitro-group to amine reductionsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:4ba5e449-aaac-4c84-9fdc-ca7df79ea294EnglishSymplectic ElementsFrontiers Media2022Ramirez, MAJoseph Srinivasan, SCleary, SETodd, PMTReeve, HAVincent, KAHydrogenase-mediated reduction of flavin mononucleotide by H2 is exploited to enable cleaner application of nitroreductase enzymes for reduction of aromatic nitro functional groups. This turns the overall reaction into a biocatalytic hydrogenation. Use of flavin-containing nitroreductases in industrial biotechnology typically relies upon NADH or NADPH as reductant, together with glucose dehydrogenase and glucose as a regeneration system for the reduced nicotinamide cofactor, with 3 equivalents of the carbon-intensive glucose required for a single 6-electron nitro to amine conversion. We show here that reduced flavin mononucleotide is an alternative reductant for nitroreductases, and by combining this with H2-driven recycling of reduced flavin, we avoid glucose, thereby enabling atom-efficient biocatalytic nitro reductions. We compare this biocatalytic system, via green chemistry metrics, to existing strategies for biocatalytic nitro-group reductions, particularly with respect to replacing glucose with H2 gas. We take steps towards demonstrating industrial viability: we report an overexpression system for E. coli hydrogenase 1, giving a 12-fold improvement in enzyme yield; we show a reaction in which the hydrogenase exhibits > 26,000 enzyme turnovers; and we demonstrate reasonable solvent tolerance of the hydrogenase and flavin reduction system which would enable reaction intensification. |
| spellingShingle | Ramirez, MA Joseph Srinivasan, S Cleary, SE Todd, PMT Reeve, HA Vincent, KA H2-driven reduction of flavin by hydrogenase enables cleaner operation of nitroreductases for nitro-group to amine reductions |
| title | H2-driven reduction of flavin by hydrogenase enables cleaner operation of nitroreductases for nitro-group to amine reductions |
| title_full | H2-driven reduction of flavin by hydrogenase enables cleaner operation of nitroreductases for nitro-group to amine reductions |
| title_fullStr | H2-driven reduction of flavin by hydrogenase enables cleaner operation of nitroreductases for nitro-group to amine reductions |
| title_full_unstemmed | H2-driven reduction of flavin by hydrogenase enables cleaner operation of nitroreductases for nitro-group to amine reductions |
| title_short | H2-driven reduction of flavin by hydrogenase enables cleaner operation of nitroreductases for nitro-group to amine reductions |
| title_sort | h2 driven reduction of flavin by hydrogenase enables cleaner operation of nitroreductases for nitro group to amine reductions |
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