Purification and characterization of the sesquiterpene cyclase aristolochene synthase from Penicillium roqueforti

Arch Biochem Biophys. 1989 Jul;272(1):137-43. doi: 10.1016/0003-9861(89)90204-x.


The sesquiterpene cyclase, aristolochene synthase, has been purified from Penicillium roqueforti by gel filtration and anion-exchange chromatography. Isolation was facilitated by a change in the elution behavior of the enzyme during gel filtration at different steps in the purification. The purified enzyme had a specific activity of 70 nmol/min/mg protein. The molecular weight as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was Mr 37,000. The native molecular weight as determined by gel filtration chromatography was Mr 48,000. The requirement for Mg2+ could be partially substituted with 0.01 mM Mn2+, but higher concentrations were inhibitory. Pyrophosphate, a competitive inhibitor of most terpene cyclases, had no effect on enzyme activity up to a concentration of 5.0 mM. The maximum activity was observed between pH 6.25 and pH 7.50, and the Km for farnesyl pyrophosphate was 0.55 +/- 0.06 microM.

MeSH terms

  • Cations
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Diphosphates / pharmacology
  • Electrophoresis, Polyacrylamide Gel
  • Hydrogen-Ion Concentration
  • Isomerases / antagonists & inhibitors
  • Isomerases / isolation & purification*
  • Isomerases / metabolism
  • Magnesium / pharmacology
  • Manganese / pharmacology
  • Molecular Weight
  • Penicillium / enzymology*
  • Polyisoprenyl Phosphates / pharmacology
  • Sesquiterpenes


  • Cations
  • Diphosphates
  • Polyisoprenyl Phosphates
  • Sesquiterpenes
  • Manganese
  • farnesyl pyrophosphate
  • Isomerases
  • aristolochene synthase
  • Magnesium