A Novel Non-Heme Iron-Containing Dioxygenase. Chloridazon-catechol Dioxygenase From Phenylobacterium Immobilis DSM 1986

Eur J Biochem. 1982 Jul;125(3):579-84. doi: 10.1111/j.1432-1033.1982.tb06722.x.

Abstract

Previously we purified an enzyme from Phenylobacterium immobilis DSM 1986, which cleaves the catechol derivative of the herbicide Chloridazon [5-amino-4-chloro-2-phenyl-3 (2H)-pyridazinone] in the meta position. The enzyme, which could be crystallized, proved in Ouchterlony double-diffusion tests to consist of a single protein species. No cross-reaction was observed with other meta-cleaving enzymes. Its light absorption spectrum showed a maximum at 279 nm (epsilon = 310 mM -1 cm -1), shoulders at 289 nm and 275 nm and a very weak band at around 430 nm (epsilon = 1.14 mM -1 cm -1). The amino acid analysis showed a slight excess of acidic amino acids, in agreement with the pl of 4.5. Surprisingly the enzyme per se is completely inactive, although it contains one non-dialysable iron atom per submit. It has to be activated by preincubation with ferrous ions or ascorbate. The enzyme activated this way is autoxidizable and returns to its non-activated state in the presence of oxygen. During the reaction with the substrate, this inactivation seems to be enhanced about 100 times. Since this kind of activation and inactivation is not observed in other meta-cleaving enzymes, this enzyme seems to represent a new type of a non-heme iron dioxygenase. We tentatively propose the name Chloridazon-catechol dioxygenase for this enzyme.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / analysis
  • Bacteria / enzymology*
  • Catechol 2,3-Dioxygenase
  • Chemical Phenomena
  • Chemistry
  • Circular Dichroism
  • Dialysis
  • Dioxygenases*
  • Immunodiffusion
  • Iron / analysis*
  • Isoelectric Focusing
  • Oxygen Consumption
  • Oxygenases / analysis*
  • Proteins / analysis
  • Spectrophotometry

Substances

  • Amino Acids
  • Proteins
  • Iron
  • Oxygenases
  • Dioxygenases
  • Catechol 2,3-Dioxygenase