Purification and Characterization of the 3-chloro-4-hydroxy-phenylacetate Reductive Dehalogenase of Desulfitobacterium Hafniense

FEBS Lett. 1998 Oct 2;436(2):159-62. doi: 10.1016/s0014-5793(98)01114-4.


The membrane-bound 3-chloro-4-hydroxyphenylacetate (Cl-OHPA) reductive dehalogenase from the chlorophenol-reducing anaerobe Desulfitobacterium hafniense was purified 11.3-fold to apparent homogeneity in the presence of the detergent CHAPS. The purified dehalogenase catalyzed the reductive dechlorination of Cl-OHPA to 4-hydroxyphenylacetate with reduced methyl viologen as the electron donor at a specific activity of 103.2 nkat/mg protein. SDS-PAGE revealed a single protein band with an apparent molecular mass of 46.5 kDa. The enzyme contained 0.68 +/- 0.2 mol corrinoid, 12.0 +/- 0.7 mol iron, and 13.0 +/- 0.7 mol acid-labile sulfur per mol subunit. The N-terminal amino acid sequence of the enzyme was determined and no significant similarity was found to any protein present in the gene bank.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / metabolism
  • Cholic Acids
  • Chromatography, Ion Exchange
  • Detergents
  • Electrophoresis, Polyacrylamide Gel
  • Gram-Negative Anaerobic Bacteria / enzymology*
  • Molecular Weight
  • Oxidoreductases / isolation & purification*
  • Oxidoreductases / metabolism*
  • Phenols / metabolism
  • Phenoxyacetates
  • Spectrophotometry
  • Substrate Specificity


  • 3-chloro-4-hydroxyphenoxyacetic acid
  • Acetates
  • Cholic Acids
  • Detergents
  • Phenols
  • Phenoxyacetates
  • Oxidoreductases
  • 3-chloro-4-hydroxyphenylacetate reductive dehalogenase
  • 3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate