The purification and characterization of 4-ethylphenol methylenehydroxylase, a flavocytochrome from Pseudomonas putida JD1.
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.
Reeve
C D
CD
Department of Biochemistry, University College of Wales, U.K.
Carver
M A
MA
Hopper
D J
DJ
D003574:Cytochrome c Group / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005420:Flavoproteins / Q000032:analysis; D006900:Hydroxylation; D007700:Kinetics; D046911:Macromolecular Substances; D006899:Mixed Function Oxygenases / Q000378:metabolism; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010636:Phenols / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology* / Q000648:ultrastructure; D013053:Spectrophotometry; D013379:Substrate Specificity
D003574:Cytochrome c Group; D005420:Flavoproteins; D046911:Macromolecular Substances; D010636:Phenols; C042291:4-ethylphenol; D006899:Mixed Function Oxygenases; C062024:4-ethylphenol methylenehydroxylase