title: Purification, crystallization and preliminary X-ray diffraction analysis of a novel keto-deoxy-D-galactarate (KDG) dehydratase from Agrobacterium tumefaciens.
abstract: D-galacturonic acid is the main component of pectin. It could be used to produce affordable renewable fuels, chemicals and materials through biotechnical conversion. Keto-deoxy-D-galactarate (KDG) dehydratase is an enzyme in the oxidative pathway of D-galacturonic acid in Agrobacterium tumefaciens (At). It converts 3-deoxy-2-keto-L-threo-hexarate to α-ketoglutaric semialdehyde. At KDG dehydratase was crystallized by the hanging-drop vapour-diffusion method. The crystals belonged to the monoclinic space group C2, with unit-cell parameters a = 169.1, b = 117.8, c = 74.3 Å, β = 112.4° and an asymmetric unit of four monomers. X-ray diffraction data were collected to 1.9 Å resolution using synchrotron radiation. The three-dimensional structure of At KDG dehydratase will provide valuable information on the function of the enzyme and will allow it to be engineered for biorefinery-based applications.
authors.affiliation: Department of Chemistry, University of Eastern Finland, PO Box 111, 80101 Joensuu, Finland.
authors.forename: Helena
authors.initials: H
authors.lastname: Taberman
authors.affiliation: VTT Technical Research Centre of Finland, PO Box 1000, 02044 VTT, Finland.
authors.forename: Martina
authors.initials: M
authors.lastname: Andberg
authors.affiliation: Department of Chemistry, University of Eastern Finland, PO Box 111, 80101 Joensuu, Finland.
authors.forename: Tarja
authors.initials: T
authors.lastname: Parkkinen
authors.affiliation: VTT Technical Research Centre of Finland, PO Box 1000, 02044 VTT, Finland.
authors.forename: Peter
authors.initials: P
authors.lastname: Richard
authors.affiliation: Department of Chemistry, University of Eastern Finland, PO Box 111, 80101 Joensuu, Finland.
authors.forename: Nina
authors.initials: N
authors.lastname: Hakulinen
authors.affiliation: VTT Technical Research Centre of Finland, PO Box 1000, 02044 VTT, Finland.
authors.forename: Anu
authors.initials: A
authors.lastname: Koivula
authors.affiliation: Department of Chemistry, University of Eastern Finland, PO Box 111, 80101 Joensuu, Finland.
authors.forename: Juha
authors.initials: J
authors.lastname: Rouvinen
mesh_terms: D016960:Agrobacterium tumefaciens / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D006836:Hydro-Lyases / Q000737:chemistry* / Q000302:isolation & purification*; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D006836:Hydro-Lyases
keywords: Agrobacterium tumefaciens; d-galacturonic acid; keto-deoxy-d-galactarate dehydratase; oxidative pathway