Structure‐Guided Modulation of the Catalytic Properties of (2Fe−2S)‐Dependent Dehydratases

Abstract: The FeS cluster‐dependent dihydroxyacid dehydratases (DHADs) and sugar acid‐specific dehydratases (DHTs) from the ilvD/EDD superfamily are key enzymes in the bioproduction of a wide variety of chemicals. We analyzed [2Fe−2S]‐dependent dehydratases in silico and in vitro, deduced functionally relevant sequence, structure, and activity relationships within the ilvD/EDD superfamily, and we propose a new classification based on their evolutionary relationships and substrate profiles. In silico simulations and analyses identified several key positions for specificity, which were experimentally investigated with site‐directed and saturation mutagenesis. We thus increased the promiscuity of DHAD from Fontimonas thermophila (FtDHAD), showing >10‐fold improved activity toward D‐gluconate, and shifted the substrate preference of DHT from Paralcaligenes ureilyticus (PuDHT) toward shorter sugar acids (recording a six‐fold improved activity toward the non‐natural substrate D‐glycerate). The successful elucidation of the role of important active site residues of the ilvD/EDD superfamily will further guide developments of this important biocatalyst for industrial applications.

Angaben zum Objekt

• Standort: Deutsche Nationalbibliothek Frankfurt am Main
• Umfang: Online-Ressource
• Sprache: Englisch

Verweise und Beziehungen

• Erschienen in: Structure‐Guided Modulation of the Catalytic Properties of (2Fe−2S)‐Dependent Dehydratases ; day:23 ; month:03 ; year:2022 ; extent:1 ; ChemBioChem ; (23.03.2022) (gesamt 1)

Beteiligte, Orts- und Zeitangaben

• Urheber: Melse, Okke ; Sutiono, Samuel ; Haslbeck, Magdalena ; Schenk, Gerhard ; Antes, Iris ; Sieber, Volker

Weitere Informationen

• DOI: 10.1002/cbic.202200088
• URN: urn:nbn:de:101:1-2022032606442605419116
• Rechteinformation: Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
• Letzte Aktualisierung: 15.08.2025, 07:27 MESZ