Purification, crystallization and preliminary X-ray diffraction studies of the three components of the toluene 2,3-dioxygenase enzyme system
- Department of Microbiology and Center for Biocatalysis and Bioprocessing, The University of Iowa, Iowa City, Iowa 52242 (United States)
- Department of Molecular Biology, Swedish University of Agricultural Sciences, Uppsala Biomedical Center, Box 590, S-751 24 Uppsala (Sweden)
All three components of the toluene dioxygenase system have been expressed, purified and crystallized. 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 Å and belongs to space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 77.1, c = 156.3 Å. Ferredoxin{sub TOL} diffracts to 1.2 Å and belongs to space group P2{sub 1}, with unit-cell parameters a = 30.5, b = 52.0, c = 30.95 Å, β = 113.7°. Apo-Oxygenase{sub TOL} and Oxygenase{sub TOL} diffract to 3.2 Å and belong to space group P4{sub 3}32, with unit-cell parameters a = 235.9 Å and a = 234.5 Å, respectively.
- OSTI ID:
- 22356144
- Journal Information:
- Acta Crystallographica. Section F, Vol. 61, Issue Pt 7; Other Information: PMCID: PMC1952458; PMID: 16511124; PUBLISHER-ID: ll5023; OAI: oai:pubmedcentral.nih.gov:1952458; Copyright (c) International Union of Crystallography 2005; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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