Expression of enzymatically active rat liver and human placental catechol-O-methyltransferase in Escherichia coli; purification and partial characterization of the enzyme

Biochim Biophys Acta. 1992 Jan 6;1129(2):149-54. doi: 10.1016/0167-4781(92)90479-j.


To produce sufficient amounts of recombinant catechol-O-methyltransferase (COMT) for structural and functional studies the coding regions of the rat liver and human placental COMT genes have been introduced into a bacterial expression vector pKEX14. Recombinant COMT was produced in Escherichia coli up to 10% of total bacterial protein after the induction of the T7 RNA polymerase gene with isopropyl-beta-D-thiogalactopyranoside. Both the rat and human enzymes were enzymatically active, soluble and reacted with anti-COMT antiserum in Western blotting. Both enzymes were purified from E. coli cells and partially characterized by determining their specific activity, apparent molecular weight and pI.

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Western
  • Catechol O-Methyltransferase / biosynthesis*
  • Catechol O-Methyltransferase / genetics
  • Catechol O-Methyltransferase / isolation & purification
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Genetic Vectors
  • Humans
  • Liver / enzymology*
  • Molecular Sequence Data
  • Placenta / enzymology*
  • Rats
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification


  • Recombinant Proteins
  • Catechol O-Methyltransferase