Open AccessPurification, crystallization and preliminary X‐ray diffraction studies of the three components of the toluene 2,3‐dioxygenase enzyme system
Author(s) -
Lee Kyoung,
Friemann Rosmarie,
Parales Juan V.,
Gibson David T.,
Ramaswamy S.
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105017549
Subject(s) - ferredoxin , dioxygenase , toluene , crystallization , pseudomonas putida , chemistry , enzyme , crystallography , stereochemistry , organic chemistry
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 (Reductase TOL ), a ferredoxin (Ferredoxin TOL ) and a Rieske non‐heme iron dioxygenase (Oxygenase TOL ). The three components and the apoenzyme of the dioxygenase (apo‐Oxygenase TOL ) were overexpressed, purified and crystallized. Reductase TOL diffracts to 1.8 Å and belongs to space group P 4 1 2 1 2, with unit‐cell parameters a = b = 77.1, c = 156.3 Å. Ferredoxin TOL diffracts to 1.2 Å and belongs to space group P 2 1 , with unit‐cell parameters a = 30.5, b = 52.0, c = 30.95 Å, β = 113.7°. Apo‐Oxygenase TOL and Oxygenase TOL diffract to 3.2 Å and belong to space group P 4 3 32, with unit‐cell parameters a = 235.9 Å and a = 234.5 Å, respectively.