Toluene monooxygenase from the fungus Cladosporium sphaerospermum

D.M.A.M. Luykx, F.X. Prenafeta-Boldu, J.A.M. de Bont

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22 Citations (Scopus)

Abstract

Assimilation of toluene by Cladosporium sphaerospermum is initially catalyzed by toluene monooxygenase (TOMO). TOMO activity was induced by adding toluene to a glucose-pregrown culture of C. sphaerospermum. The corresponding microsomal enzyme needed NADPH and O2 to oxidize toluene and glycerol, EDTA, DTT, and PMSF for stabilization. TOMO activity was maximal at 35 °C and pH 7.5 and was inhibited by carbon monoxide, Metyrapone, and cytochrome c. TOMO preferred as substrates also other aromatic hydrocarbons with a short aliphatic side chain. Its reduced carbon monoxide difference spectrum showed a maximum at 451 nm. A substrate-induced Type I spectrum was observed on addition of toluene. These results indicated that TOMO is a cytochrome P450. TOMO and its corresponding reductase were eventually purified by a simultaneous purification revealing apparent molecular masses of 58 and 78 kDa, respectively.
Original languageEnglish
Pages (from-to)373-379
JournalBiochemical and Biophysical Research Communications
Volume312
Issue number2
DOIs
Publication statusPublished - 2003

Keywords

  • cytochrome-c reductase
  • cytosolic cytochrome-p450
  • aromatic-hydrocarbons
  • liver microsomes
  • energy-source
  • sole carbon
  • purification
  • metabolism
  • hydroxylation
  • pathway

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