title: Structural and functional insights into asymmetric enzymatic dehydration of alkenols.
abstract: The asymmetric dehydration of alcohols is an important process for the direct synthesis of alkenes. We report the structure and substrate specificity of the bifunctional linalool dehydratase isomerase (LinD) from the bacterium Castellaniella defragrans that catalyzes in nature the hydration of β-myrcene to linalool and the subsequent isomerization to geraniol. Enzymatic kinetic resolutions of truncated and elongated aromatic and aliphatic tertiary alcohols (C5-C15) that contain a specific signature motif demonstrate the broad substrate specificity of LinD. The three-dimensional structure of LinD from Castellaniella defragrans revealed a pentamer with active sites at the protomer interfaces. Furthermore, the structure of LinD in complex with the product geraniol provides initial mechanistic insights into this bifunctional enzyme. Site-directed mutagenesis confirmed active site amino acid residues essential for its dehydration and isomerization activity. These structural and mechanistic insights facilitate the development of hydrating catalysts, enriching the toolbox for novel bond-forming biocatalysis.
authors.affiliation: Institute of Technical Biochemistry, Universitaet Stuttgart, Stuttgart, Germany.
authors.forename: Bettina M
authors.initials: BM
authors.lastname: Nestl
authors.affiliation: Institute of Technical Biochemistry, Universitaet Stuttgart, Stuttgart, Germany.
authors.forename: Christopher
authors.initials: C
authors.lastname: Geinitz
authors.affiliation: York Structural Biology Laboratory, Department of Chemistry, University of York, Heslington, York, UK.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Popa
authors.affiliation: Genomatica, Inc., San Diego, California, USA.
authors.forename: Sari
authors.initials: S
authors.lastname: Rizek
authors.affiliation: Genomatica, Inc., San Diego, California, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Haselbeck
authors.affiliation: Genomatica, Inc., San Diego, California, USA.
authors.forename: Rosary
authors.initials: R
authors.lastname: Stephen
authors.affiliation: Genomatica, Inc., San Diego, California, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Noble
authors.affiliation: Institute of Technical Biochemistry, Universitaet Stuttgart, Stuttgart, Germany.
authors.forename: Max-Philipp
authors.initials: MP
authors.lastname: Fischer
authors.affiliation: Genomatica, Inc., San Diego, California, USA.
authors.forename: Erik C
authors.initials: EC
authors.lastname: Ralph
authors.affiliation: York Structural Biology Laboratory, Department of Chemistry, University of York, Heslington, York, UK.
authors.forename: Hoi Ting
authors.initials: HT
authors.lastname: Hau
authors.affiliation: York Structural Biology Laboratory, Department of Chemistry, University of York, Heslington, York, UK.
authors.forename: Henry
authors.initials: H
authors.lastname: Man
authors.affiliation: York Structural Biology Laboratory, Department of Chemistry, University of York, Heslington, York, UK.
authors.forename: Muhiadin
authors.initials: M
authors.lastname: Omar
authors.affiliation: York Structural Biology Laboratory, Department of Chemistry, University of York, Heslington, York, UK.
authors.forename: Johan P
authors.identifier: 0000-0001-6992-6838
authors.initials: JP
authors.lastname: Turkenburg
authors.affiliation: Genomatica, Inc., San Diego, California, USA.
authors.forename: Stephen
authors.initials: S
authors.lastname: van Dien
authors.affiliation: Genomatica, Inc., San Diego, California, USA.
authors.forename: Stephanie J
authors.initials: SJ
authors.lastname: Culler
authors.affiliation: York Structural Biology Laboratory, Department of Chemistry, University of York, Heslington, York, UK.
authors.forename: Gideon
authors.initials: G
authors.lastname: Grogan
authors.affiliation: Institute of Technical Biochemistry, Universitaet Stuttgart, Stuttgart, Germany.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Hauer
mesh_terms: D000438:Alcohols / Q000737:chemistry* / Q000378:metabolism*; D055162:Biocatalysis; D003681:Dehydration; D006836:Hydro-Lyases / Q000378:metabolism*; D015394:Molecular Structure
chemical_list: D000438:Alcohols; D006836:Hydro-Lyases; C561837:linalool dehydratase-isomerase, Castellaniella defragrans