Open AccessCrystals of DhaA mutants from Rhodococcus rhodochrous NCIMB 13064 diffracted to ultrahigh resolution: crystallization and preliminary diffraction analysis
Author(s) -
Stsiapanava Alena,
Koudelakova Tana,
Lapkouski Mikalai,
Pavlova Martina,
Damborsky Jiri,
Kuta Smatanova Ivana
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108002066
Subject(s) - rhodococcus rhodochrous , crystallization , resolution (logic) , diffraction , crystallography , materials science , rhodococcus , chemistry , physics , optics , biochemistry , organic chemistry , computer science , artificial intelligence , enzyme
The enzyme DhaA from Rhodococcus rhodochrous NCIMB 13064 belongs to the haloalkane dehalogenases, which catalyze the hydrolysis of haloalkanes to the corresponding alcohols. The haloalkane dehalogenase DhaA and its variants can be used to detoxify the industrial pollutant 1,2,3‐trichloropropane (TCP). Three mutants named DhaA04, DhaA14 and DhaA15 were constructed in order to study the importance of tunnels connecting the buried active site with the surrounding solvent to the enzymatic activity. All protein mutants were crystallized using the sitting‐drop vapour‐diffusion method. The crystals of DhaA04 belonged to the orthorhombic space group P 2 1 2 1 2 1 , while the crystals of the other two mutants DhaA14 and DhaA15 belonged to the triclinic space group P 1. Native data sets were collected for the DhaA04, DhaA14 and DhaA15 mutants at beamline X11 of EMBL, DESY, Hamburg to the high resolutions of 1.30, 0.95 and 1.15 Å, respectively.