Purification, crystallization and preliminary crystallographic analysis of DehIVa, a dehalogenase from Burkholderia cepacia MBA4.
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.
Schmidberger
Jason W
JW
Structural Biology and Bioinformatics Group, School of Pharmacology and Medicine, University Of Western Australia, Crawley, Western Australia, Australia. jwschmidberger@yahoo.com.au
Oakley
Aaron J
AJ
Tsang
Jimmy S H
JS
Wilce
Matthew C J
MC
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
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D006867:Hydrolases; C034507:2-haloacid dehalogenase