title: Purification, crystallization and preliminary crystallographic analysis of DehIVa, a dehalogenase from Burkholderia cepacia MBA4.
abstract: DehIVa is one of two dehalogenases produced by the soil- and water-borne bacterium Burkholderia cepacia MBA4. It acts to break down short-chain halogenated aliphatic acids through a nucleophilic attack and subsequent hydrolysis of an enzyme-substrate intermediate to remove the halide ions from L-enantiomers substituted at the C2 position (e.g L-2-monochloropropionic acid). Dehalogenases are an important group of enzymes that are responsible for breaking down a diverse range of halogenated environmental pollutants. The dhlIVa gene coding for DehIVa was expressed in Escherichia coli and the protein was purified and crystallized using the hanging-drop method. Crystals grown in PEG 4000 and ammonium sulfate diffracted to 3.1 A. The crystals had a primitive hexagonal unit cell, with unit-cell parameters a = b = 104.2, c = 135.8 A, alpha = beta = 90, gamma = 120 degrees. Determining this structure will provide valuable insights into the characterization of the catalytic mechanisms of this group of enzymes.
authors.affiliation: Structural Biology and Bioinformatics Group, School of Pharmacology and Medicine, University Of Western Australia, Crawley, Western Australia, Australia. jwschmidberger@yahoo.com.au
authors.forename: Jason W
authors.initials: JW
authors.lastname: Schmidberger
authors.forename: Aaron J
authors.initials: AJ
authors.lastname: Oakley
authors.forename: Jimmy S H
authors.initials: JS
authors.lastname: Tsang
authors.forename: Matthew C J
authors.initials: MC
authors.lastname: Wilce
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D016956:Burkholderia cepacia / Q000201:enzymology*; D003460:Crystallization; D006867:Hydrolases / Q000737:chemistry* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D006867:Hydrolases; C034507:2-haloacid dehalogenase