Premium
Structure‐Based Mechanism of Oleate Hydratase from Elizabethkingia meningoseptica
Author(s) -
Engleder Matthias,
PavkovKeller Tea,
Emmerstorfer Anita,
Hromic Altijana,
Schrempf Sabine,
Steinkellner Georg,
Wriessnegger Tamara,
Leitner Erich,
Strohmeier Gernot A.,
Kaluzna Iwona,
Mink Daniel,
Schürmann Martin,
Wallner Silvia,
Macheroux Peter,
Gruber Karl,
Pichler Harald
Publication year - 2015
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500269
Subject(s) - chemistry , cofactor , flavoprotein , stereochemistry , protonation , flavin group , double bond , substrate (aquarium) , flavin adenine dinucleotide , active site , enzyme , biochemistry , biology , organic chemistry , ion , ecology
Hydratases provide access to secondary and tertiary alcohols by regio‐ and/or stereospecifically adding water to carbon‐carbon double bonds. Thereby, hydroxy groups are introduced without the need for costly cofactor recycling, and that makes 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 flavin adenine dinucleotide (FAD). A structure‐based mutagenesis study targeting active site residues identified E122 and Y241 as crucial for the activation of a water molecule and for protonation of the double bond, respectively. Moreover, we also observed that two‐electron reduction of FAD results in a sevenfold increase in the substrate hydration rate. We propose the first reaction mechanism for this enzyme class that explains the requirement for the flavin cofactor and the involvement of conserved amino acid residues in this regio‐ and stereoselective hydration.