The dodecameric vanadium-dependent haloperoxidase from the marine algae Corallina officinalis: cloning, expression, and refolding of the recombinant enzyme

Protein Expr Purif. 2007 Apr;52(2):265-72. doi: 10.1016/j.pep.2006.08.010. Epub 2006 Aug 30.

Abstract

The dodecameric vanadium-dependent bromoperoxidase from Corallina officinalis has been cloned and over-expressed in Escherichia coli. However, the enzyme was found to be predominantly in the form of inclusion bodies. This protein presents a challenging target for refolding, both due to the size (768kDa) and quaternary structure (12x64kDa). Successful refolding conditions have been established which result in an increase in the final yield of active bromoperoxidase from 0.5mg to 40mg per litre of culture. The refolded protein has been characterised and compared to the native enzyme and was shown to be stable at temperatures of 80 degrees C, over a pH range 5.5-10 and in organic solvents such as ethanol, acetonitrile, methanol, and acetone. The novel refolding approach reported in this paper opens up the full potential of this versatile enzyme for use in large scale biotransformation studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Drug Combinations
  • Escherichia coli / genetics
  • Eukaryota / enzymology*
  • Gene Expression
  • Iodide Peroxidase / chemistry*
  • Iodide Peroxidase / genetics
  • Iodide Peroxidase / metabolism
  • Marine Biology
  • Molecular Sequence Data
  • Oils
  • Phenols
  • Polymers
  • Protein Folding
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Temperature

Substances

  • Drug Combinations
  • Oils
  • P & S Liquid
  • Phenols
  • Polymers
  • Recombinant Proteins
  • vanadium iodoperoxidase
  • Iodide Peroxidase