Purification, crystallization and preliminary X-ray diffraction studies of the three components of the toluene 2,3-dioxygenase enzyme system

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Jul 1;61(Pt 7):669-72. doi: 10.1107/S1744309105017549. Epub 2005 Jun 15.

Abstract

Pseudomonas putida F1 can grow with toluene as its sole source of carbon and energy. The initial reaction of the degradation of toluene is catalyzed by a three-component toluene dioxygenase enzyme system consisting of a reductase (ReductaseTOL), a ferredoxin (FerredoxinTOL) and a Rieske non-heme iron dioxygenase (OxygenaseTOL). The three components and the apoenzyme of the dioxygenase (apo-OxygenaseTOL) were overexpressed, purified and crystallized. ReductaseTOL diffracts to 1.8 A and belongs to space group P4(1)2(1)2, with unit-cell parameters a = b = 77.1, c = 156.3 A. Ferredoxin(TOL) diffracts to 1.2 A and belongs to space group P2(1), with unit-cell parameters a = 30.5, b = 52.0, c = 30.95 A, beta = 113.7 degrees. Apo-OxygenaseTOL and OxygenaseTOL diffract to 3.2 A and belong to space group P4(3)32, with unit-cell parameters a = 235.9 A and a = 234.5 A, respectively.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Crystallization
  • Crystallography, X-Ray
  • Dioxygenases / chemistry
  • Ferredoxins / chemistry
  • Mixed Function Oxygenases / chemistry*
  • Models, Biological
  • Models, Chemical
  • Pseudomonas putida / enzymology*
  • Temperature
  • X-Ray Diffraction

Substances

  • Ferredoxins
  • Mixed Function Oxygenases
  • Dioxygenases
  • toluene 2,3-dioxygenase