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

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Kyoung LeeS Ramaswamy

Abstract

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.

References

Dec 14, 1979·Biochemical and Biophysical Research Communications·V SubramanianD T Gibson
Jan 1, 1997·Environmental Research·M M Greenberg
Oct 26, 1999·Acta Crystallographica. Section D, Biological Crystallography·H R Powell
Oct 26, 1999·Acta Crystallographica. Section D, Biological Crystallography·J W Pflugrath
Jun 14, 2000·Current Opinion in Biotechnology·D T Gibson, R E Parales

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Citations

Jan 21, 2009·Acta Crystallographica. Section D, Biological Crystallography·Rosmarie FriemannS Ramaswamy
Feb 1, 2008·Expert Opinion on Drug Discovery·Ramesh N Patel
Sep 20, 2005·Biochemical and Biophysical Research Communications·Daniel J FerraroS Ramaswamy
Nov 26, 2014·The Journal of General and Applied Microbiology·Ajiraporn KongpolAlisa S Vangnai

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