Characterisation of an L-haloacid dehalogenase from the marine psychrophile Psychromonas ingrahamii with potential industrial application

Mar Biotechnol (NY). 2013 Dec;15(6):695-705. doi: 10.1007/s10126-013-9522-3. Epub 2013 Aug 16.

Abstract

The recombinant L-haloacid dehalogenase from the marine bacterium Psychromonas ingrahamii has been cloned and over-expressed in Escherichia coli. It shows activity towards monobromoacetic (100 %), monochloroacetic acid (62 %), S-chloropropionic acid (42 %), S-bromopropionic acid (31 %), dichloroacetic acid (28 %) and 2-chlorobutyric acid (10 %), respectively. The L-haloacid dehalogenase has highest activity towards substrates with shorter carbon chain lengths (≤ C3), without preference towards a chlorine or bromine at the α-carbon position. Despite being isolated from a psychrophilic bacterium, the enzyme has mesophilic properties with an optimal temperature for activity of 45 °C. It retains above 70 % of its activity after being incubated at 65 °C for 90 min before being assayed at 25 °C. The enzyme is relatively stable in organic solvents as demonstrated by activity and thermal shift analysis. The V max and K m were calculated to be 0.6 μM min(-1) mg(-1) and 1.36 mM with monobromoacetic acid, respectively. This solvent-resistant and stable L-haloacid dehalogenase from P. ingrahamii has potential to be used as a biocatalyst in industrial processes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / metabolism
  • Amino Acid Sequence
  • Base Sequence
  • Biocatalysis
  • Cloning, Molecular
  • Computational Biology
  • DNA Primers / genetics
  • Dichloroacetic Acid / metabolism
  • Escherichia coli
  • Gammaproteobacteria / enzymology*
  • Hydrolases / genetics*
  • Hydrolases / metabolism*
  • Industrial Microbiology / methods*
  • Models, Molecular*
  • Molecular Sequence Data
  • Propionates / metabolism
  • Protein Conformation*
  • Sequence Analysis, DNA
  • Temperature

Substances

  • Acetates
  • DNA Primers
  • Propionates
  • chloroacetic acid
  • bromoacetate
  • Dichloroacetic Acid
  • Hydrolases
  • 2-haloacid dehalogenase
  • propionic acid