Open AccessPurification, crystallization and X‐ray diffraction analysis of a novel ring‐cleaving enzyme (BoxC C ) from Burkholderia xenovorans LB400
Author(s) -
Bains Jasleen,
Boulanger Martin J.
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108010919
Subject(s) - crystallization , ring (chemistry) , crystallography , enzyme , burkholderia , x ray crystallography , materials science , chemistry , diffraction , biochemistry , organic chemistry , physics , biology , bacteria , optics , genetics
The assimilation of aromatic compounds by microbial species requires specialized enzymes to cleave the thermodynamically stable ring. In the recently discovered benzoate‐oxidation ( box ) pathway in Burkholderia xenovorans LB400, this is accomplished by a novel dihydrodiol lyase (BoxC C ). Sequence analysis suggests that BoxC C is part of the crotonase superfamily but includes an additional uncharacterized region of approximately 115 residues that is predicted to mediate ring cleavage. Processing of X‐ray diffraction data to 1.5 Å resolution revealed that BoxC C crystallized with two molecules in the asymmetric unit of the P 2 1 2 1 2 1 space group, with a solvent content of 47% and a Matthews coefficient of 2.32 Å 3 Da −1 . Selenomethionine BoxC C has been purified and crystals are currently being refined for anomalous dispersion studies.