Purification, crystallization and preliminary X-ray diffraction studies of the three components of the toluene 2,3-dioxygenase enzyme system.
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.
Lee
Kyoung
K
Department of Microbiology, Center for Biocatalysis and Bioprocessing, The University of Iowa, Iowa City, Iowa 52242, USA.
Friemann
Rosmarie
R
Parales
Juan V
JV
Gibson
David T
DT
Ramaswamy
S
S
D003460:Crystallization; D018360:Crystallography, X-Ray; D049308:Dioxygenases / Q000737:chemistry; D005288:Ferredoxins / Q000737:chemistry; D006899:Mixed Function Oxygenases / Q000737:chemistry*; D008954:Models, Biological; D008956:Models, Chemical; D016958:Pseudomonas putida / Q000201:enzymology*; D013696:Temperature; D014961:X-Ray Diffraction
D005288:Ferredoxins; D006899:Mixed Function Oxygenases; D049308:Dioxygenases; C112823:toluene 2,3-dioxygenase