Open AccessCrystallization and preliminary X‐ray diffraction analysis of recombinant chlorocatechol 1,2‐dioxygenase from Pseudomonas putida
Author(s) -
Rustiguel Joane Kathelen,
Pinheiro Matheus Pinto,
Araújo Ana Paula Ulian,
ato Maria Cristina
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430911100635x
Subject(s) - pseudomonas putida , crystallization , monoclinic crystal system , dioxygenase , rhodococcus , crystallography , molecule , crystal structure , materials science , polyethylene glycol , chemistry , organic chemistry , enzyme
Chlorocatechol 1,2‐dioxygenase from the Gram‐negative bacterium Pseudomonas putida (Pp 1,2‐CCD) is considered to be an important biotechnological tool owing to its ability to process a broad spectrum of organic pollutants. In the current work, the crystallization, crystallographic characterization and phasing of the recombinant Pp 1,2‐CCD enzyme are described. Reddish‐brown crystals were obtained in the presence of polyethylene glycol and magnesium acetate by utilizing the vapour‐diffusion technique in sitting drops. Crystal dehydration was the key step in obtaining data sets, which were collected on the D03B‐MX2 beamline at the CNPEM/MCT – LNLS using a MAR CCD detector. Pp 1,2‐CCD crystals belonged to space group P 6 1 22 and the crystallographic structure of Pp 1,2‐CCD has been solved by the MR‐SAD technique using Fe atoms as scattering centres and the coordinates of 3‐chlorocatechol 1,2‐dioxygenase from Rhodococcus opacus (PDB entry 2boy ) as the search model. The initial model, which contains three molecules in the asymmetric unit, has been refined to 3.4 Å resolution.