Open AccessPurification, crystallization and preliminary crystallographic analysis of DehIVa, a dehalogenase from Burkholderia cepacia MBA4
Author(s) -
Schmidberger Jason W.,
Oakley Aaron J.,
Tsang Jimmy S. H.,
Wilce Matthew C. J.
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105002472
Subject(s) - crystallization , dehalogenase , burkholderia , crystallography , chemistry , microbiology and biotechnology , biology , bacteria , biochemistry , organic chemistry , enzyme , genetics
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, α = β = 90, γ = 120°. Determining this structure will provide valuable insights into the characterization of the catalytic mechanisms of this group of enzymes.