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Publication Abstract from PubMed

Hydratases provide access to secondary and tertiary alcohols by regio- and/or stereospecifically adding water to carbon-carbon double bonds. Thereby, hydroxyl groups are introduced without the need for costly co-factor recycling, which renders this approach highly interesting on an industrial scale. Here, we present the first crystal structure of a recombinant oleate hydratase originating from Elizabethkingia meningoseptica in the presence of the flavin cofactor (FAD). A structure-based mutagenesis study targeting active site residues identified E122 and Y241 as crucial for the activation of a water molecule and protonation of the double bond, respectively. Moreover, we also observed that two-electron reduction of FAD results in a 7-fold increase of substrate hydration rate. We propose the first reaction mechanism for this enzyme class that rationalizes the requirement of flavin cofactor and the involvement of conserved amino acid residues in this regio- and stereoselective hydration.

Structure-based mechanism of oleate hydratase from Elizabethkingia meningoseptica.,Engleder M, Pavkov-Keller T, Emmerstorfer A, Hromic A, Schrempf S, Steinkellner G, Wriessnegger T, Leitner E, Strohmeier GA, Kaluzna I, Mink D, Schurmann M, Wallner S, Macheroux P, Gruber K, Pichler H Chembiochem. 2015 Jun 15. doi: 10.1002/cbic.201500269. PMID:26077980^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.