Acta Crystallographica Section F

Structural Biology and Crystallization Communications

Volume 61, Part 3 (March 2005)

crystallization communications

Acta Cryst. (2005). F61, 271-273 [ doi:10.1107/S1744309105002472 ]

Purification, crystallization and preliminary crystallographic analysis of DehIVa, a dehalogenase from Burkholderia cepacia MBA4

J. W. Schmidberger, A. J. Oakley, J. S. H. Tsang and M. C. J. Wilce

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 Å. The crystals had a primitive hexagonal unit cell, with unit-cell parameters a = b = 104.2, c = 135.8 Å, [alpha] = [beta] = 90, [gamma] = 120°. Determining this structure will provide valuable insights into the characterization of the catalytic mechanisms of this group of enzymes.

Keywords: dehalogenases; Burkholderia cepacia MBA4; DehIVa; halogenated organic acids.