Crystallization and preliminary characterization of chloromuconolactone dehalogenase from Rhodococcus opacus 1CP.
Chloroaromatic compounds are often very persistent environmental pollutants. Nevertheless, numerous bacteria are able to metabolize these compounds and to utilize them as sole energy and carbon sources. Rhodococcus opacus 1CP is able to degrade several chloroaromatic compounds, some of them via a variation of the 3-chlorocatechol branch of the modified ortho-cleavage pathway. This branch in R. opacus differs from that in Proteobacteria in the inability of the chloromuconate cycloisomerase to dehalogenate. Instead, a unique enzyme designated as chloromuconolactone dehalogenase (ClcF) is recruited. ClcF dehalogenates 5-chloromuconolactone to cis-dienelactone and shows a high similarity to muconolactone isomerases (EC 5.3.3.4). However, unlike the latter enzymes, it is unable to catalyse the isomerization of muconolactone to 3-oxoadipate enollactone. In order to characterize the catalytic mechanism of this unusual dehalogenase, the enzyme was crystallized and subjected to X-ray structural analysis. Data sets to up to 1.65 Å resolution were collected from two different crystal forms using synchrotron radiation. Crystal form I (space group P2(1)) contained 40 subunits in the asymmetric unit, whereas ten subunits were present in crystal form II (space group P2(1)2(1)2(1)). The self-rotation function revealed the orientations of the molecular symmetry axes of the homodecamer of 52 symmetry.
Center for Biotechnology and Biomedicine, Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, University of Leipzig, Deutscher Platz 5, 04103 Leipzig, Germany.
Christian
C
Roth
Stefan R
SR
Kaschabek
Janosch A D
JA
Gröning
Thomas
T
Handrek
Michael
M
Schlömann
Norbert
N
Sträter
D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D003460:Crystallization; D006867:Hydrolases / Q000737:chemistry* / Q000378:metabolism; D012240:Rhodococcus / Q000201:enzymology*
C088142:5-chloromuconolactone; D006867:Hydrolases; D015107:4-Butyrolactone