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; pp. 669 - 672
• Main Authors: Lee, Kyoung, Friemann, Rosmarie, Parales, Juan V., Gibson, David T., Ramaswamy, S.
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Munksgaard International Publishers 01.07.2005; International Union of Crystallography
• Subjects: 3-dioxygenase enzyme system; CARBON; CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; Crystallization Communications; Crystallography, X-Ray; Dioxygenases - chemistry; Ferredoxins - chemistry; HEME; IRON; Mixed Function Oxygenases - chemistry; Models, Biological; Models, Chemical; OXYGEN; Pseudomonas putida - enzymology; SPACE GROUPS; Temperature; TOLUENE; toluene 2; toluene 2,3‐dioxygenase enzyme system; X-RAY DIFFRACTION
• ISSN: 1744-3091; 1744-3091
• DOI: 10.1107/S1744309105017549

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 Å and belongs to space group P41212, with unit‐cell parameters a = b = 77.1, c = 156.3 Å. FerredoxinTOL diffracts to 1.2 Å and belongs to space group P21, with unit‐cell parameters a = 30.5, b = 52.0, c = 30.95 Å, β = 113.7°. Apo‐OxygenaseTOL and OxygenaseTOL diffract to 3.2 Å and belong to space group P4332, with unit‐cell parameters a = 235.9 Å and a = 234.5 Å, respectively.