Purification and characterization of haloalcohol dehalogenase from Arthrobacter sp. strain AD2

J Bacteriol. 1991 Jan;173(1):124-9. doi: 10.1128/jb.173.1.124-129.1991.

Abstract

An enzyme capable of dehalogenating vicinal haloalcohols to their corresponding epoxides was purified from the 3-chloro-1,2-propanediol-utilizing bacterium Arthrobacter sp. strain AD2. The inducible haloalcohol dehalogenase converted 1,3-dichloro-2-propanol, 3-chloro-1,2-propanediol, 1-chloro-2-propanol, and their brominated analogs, 2-bromoethanol, as well as chloroacetone and 1,3-dichloroacetone. The enzyme possessed no activity for epichlorohydrin (3-chloro-1,2-epoxypropane) or 2,3-dichloro-1-propanol. The dehalogenase had a broad pH optimum at about 8.5 and a temperature optimum of 50 degrees C. The enzyme followed Michaelis-Menten kinetics, and the Km values for 1,3-dichloro-2-propanol and 3-chloro-1,2-propanediol were 8.5 and 48 mM, respectively. Chloroacetic acid was a competitive inhibitor, with a Ki of 0.50 mM. A subunit molecular mass of 29 kDa was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. With gel filtration, a molecular mass of 69 kDa was found, indicating that the native protein is a dimer. The amino acid composition and N-terminal amino acid sequence are given.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Arthrobacter / enzymology*
  • Blotting, Western
  • Chromatography
  • Chromatography, DEAE-Cellulose
  • Chromatography, Gel
  • Durapatite
  • Electrophoresis, Polyacrylamide Gel
  • Hydroxyapatites
  • Kinetics
  • Lyases / genetics
  • Lyases / isolation & purification*
  • Lyases / metabolism
  • Molecular Sequence Data
  • Molecular Weight
  • Substrate Specificity

Substances

  • Hydroxyapatites
  • Durapatite
  • Lyases
  • haloalcohol hydrogen-halide lyase