Scanning aldoxime dehydratase sequence space and characterization of a new aldoxime dehydratase from Fusarium vanettenii

0569334 - MBÚ 2024 RIV NL eng J - Článek v odborném periodiku
Křístková, Barbora - Rädisch, Robert - Kulik, Natalia - Horvat, M. - Rucká, Lenka - Grulich, Michal - Rudroff, F. - Kádek, Alan - Pátek, Miroslav - Winkler, M. - Martínková, Ludmila
Scanning aldoxime dehydratase sequence space and characterization of a new aldoxime dehydratase from Fusarium vanettenii.
Enzyme and Microbial Technology. Roč. 164, MAR 2023 (2023), č. článku 110187. ISSN 0141-0229. E-ISSN 1879-0909
Grant CEP: GA ČR(CZ) GF20-23532L
Institucionální podpora: RVO:61388971
Klíčová slova: Aldoxime dehydratase * Fusarium vanettenii * Catalytic triad * Nitrile synthesis * Homology modeling * Substrate docking
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 3.4, rok: 2022
Způsob publikování: Omezený přístup
https://www.sciencedirect.com/science/article/pii/S014102292200206X?via%3Dihub

The aim of this work was to map the sequence space of aldoxime dehydratases (Oxds) as enzymes with great potential for nitrile synthesis. Microbes contain an abundance of putative Oxds but fewer than ten Oxds were characterized in total and only two in fungi. In this work, we prepared and characterized a new Oxd (protein gb| EEU37245.1 named OxdFv) from Fusarium vanettenii 77-13-4. OxdFv is distant from the characterized Oxds with a maximum of 36% identity. Moreover, the canonical Oxd catalytic triad RSH is replaced by R141-E187-E303 in OxdFv. R141A and E187A mutants did not show significant activities, but mutant E303A showed a comparable activity as the wild-type enzyme. According to native mass spectrometry, OxdFv contained almost 1 mol of heme per 1 mol of protein, and was composed of approximately 88% monomer (41.8 kDa) and 12% dimer. A major advantage of this enzyme is its considerable activity under aerobic conditions (25.0 +/- 4.3 U/mg for E,Z-phe-nylacetaldoxime at pH 9.0 and 55 degrees C). Addition of sodium dithionite (reducing agent) and Fe2+ was required for this activity. OxdFv favored (aryl)aliphatic aldoximes over aromatic aldoximes. Substrate docking in the ho-mology model of OxdFv showed a similar substrate specificity. We conclude that OxdFv is the first characterized Oxd of the REE type.
Trvalý link: https://hdl.handle.net/11104/0342563