The metabolism of benzene by bacteria. Purification and some properties of the enzyme cis-1,2-dihydroxycyclohexa-3,5-diene (nicotinamide adenine dinucleotide) oxidoreductase (cis-benzene glycol dehydrogenase)

Biochem J. 1973 Dec;136(4):927-34. doi: 10.1042/bj1360927.

Abstract

1. cis-Benzene glycol dehydrogenase was purified to a homogeneous state from a species of Pseudomonas grown with benzene as the major carbon source. 2. The enzyme was specific for the cis-isomer of its substrate and required NAD(+) as hydrogen acceptor. 3. Partial inactivation of the enzyme, which was observed during purification, could be reversed by the addition of Fe(2+) and GSH. 4. A molecular weight of 440000 was calculated from data obtained by sedimentation-velocity and diffusion analysis in the ultracentrifuge. Sodium dodecyl sulphate polyacrylamide-gel electrophoresis indicated a subunit of molecular weight 110000. 5. p-Chloromercuribenzoic acid and 1,10-phenanthroline were shown to inhibit the enzyme.

MeSH terms

  • Alcohol Oxidoreductases / antagonists & inhibitors
  • Benzene / metabolism*
  • Cell-Free System
  • Chloromercuribenzoates / pharmacology
  • Chromatography, DEAE-Cellulose
  • Cyclohexanols
  • Diffusion
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Activation
  • Glutathione / pharmacology
  • Hydrogen-Ion Concentration
  • Iron / pharmacology
  • Isomerism
  • Molecular Weight
  • NAD
  • Oxidoreductases / isolation & purification*
  • Oxidoreductases Acting on CH-CH Group Donors
  • Phenanthrolines / pharmacology
  • Pseudomonas / enzymology*
  • Sodium Dodecyl Sulfate
  • Ultracentrifugation

Substances

  • Chloromercuribenzoates
  • Cyclohexanols
  • Phenanthrolines
  • NAD
  • Sodium Dodecyl Sulfate
  • Iron
  • Oxidoreductases
  • Alcohol Oxidoreductases
  • Oxidoreductases Acting on CH-CH Group Donors
  • cis-1,2-dihydrobenzene-1,2-diol dehydrogenase
  • Glutathione
  • Benzene