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

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 61; no. (7) ; 2005
• Main Authors: Lee, Kyoung, Friemann, Rosmarie, Parales, Juan V., Gibsona, David T., Ramaswamy, S.
• Format: Journal Article
• Language: English
• Published: United States 19.07.2010
• Subjects: 60 APPLIED LIFE SCIENCES; BASIC BIOLOGICAL SCIENCES; CARBON; CRYSTALLIZATION; ENZYMES; FERREDOXIN; IRON; OXIDOREDUCTASES; PSEUDOMONAS; PURIFICATION; SPACE GROUPS; TOLUENE; X-RAY DIFFRACTION
• ISSN: 1744-3091; 1744-3091

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{sub TOL}), a ferredoxin (Ferredoxin{sub TOL}) and a Rieske non-heme iron dioxygenase (Oxygenase{sub TOL}). The three components and the apoenzyme of the dioxygenase (apo-Oxygenase{sub TOL}) were overexpressed, purified and crystallized. Reductase{sub TOL} diffracts to 1.8 {angstrom} and belongs to space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 77.1, c = 156.3 {angstrom}. Ferredoxin{sub TOL} diffracts to 1.2 {angstrom} and belongs to space group P2{sub 1}, with unit-cell parameters a = 30.5, b = 52.0, c = 30.95 {angstrom}, {beta} = 113.7{sup o}. Apo-Oxygenase{sub TOL} and Oxygenase{sub TOL} diffract to 3.2 {angstrom} and belong to space group P4{sub 3}32, with unit-cell parameters a = 235.9 {angstrom} and a = 234.5 {angstrom}, respectively.