The purification and characterization of 4-ethylphenol methylenehydroxylase, a flavocytochrome from Pseudomonas putida JD1

Biochem J. 1989 Oct 15;263(2):431-7. doi: 10.1042/bj2630431.

Abstract

The enzyme 4-ethylphenol methylenehydroxylase was purified from Pseudomonas putida JD1 grown on 4-ethylphenol. It is a flavocytochrome c for which the Mr was found to be 120,000 by ultracentrifuging and 126,000 by gel filtration. The enzyme consists of two flavoprotein subunits each of Mr 50,000 and two cytochrome c subunits each of Mr 10,000. The redox potential of the cytochrome is 240 mV. Hydroxylation proceeds by dehydrogenation and hydration to give 1-(4'-hydroxyphenyl)ethanol, which is also dehydrogenated by the same enzyme to 4-hydroxyacetophenone. The enzyme will hydroxylate p-cresol but is more active with alkylphenols with longer-chain alkyl groups. It is located in the periplasm of the bacterium.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cytochrome c Group / analysis
  • Electrophoresis, Polyacrylamide Gel
  • Flavoproteins / analysis
  • Hydroxylation
  • Kinetics
  • Macromolecular Substances
  • Mixed Function Oxygenases / metabolism
  • Molecular Weight
  • Oxidation-Reduction
  • Phenols / metabolism
  • Pseudomonas / enzymology*
  • Pseudomonas / ultrastructure
  • Spectrophotometry
  • Substrate Specificity

Substances

  • Cytochrome c Group
  • Flavoproteins
  • Macromolecular Substances
  • Phenols
  • 4-ethylphenol
  • Mixed Function Oxygenases
  • 4-ethylphenol methylenehydroxylase