A novel haloalkane dehalogenase DgaA, isolated from Glaciecola Agarilytica NO2 was successfully crystallized.
Haloalkane dehalogenases represent an important class of microbial enzymes that catalyse an irreversible hydrolysis of halogenated compounds to the corresponding alcohol, halide ion and proton [1]. This enzymes are widely used for biodegradation and biosensing of environmental pollutants, decontamination of warfare agents, recycling by-products from chemical processes [2].
DgaA was isolated from psychrophilic and moderate halophillic organism Glaciecola Agarilytica NO2, found in the East Sea marine sediment in Korea [3].
In order to obtain the structure of DgaA x-ray diffraction analysis of enzyme's crystals was employed. Initial screening for crystallization conditions has been performed on Gryphon crystallization robot (Art Robbins Instruments, USA) using sitting drop vapour diffusion method [4]. After initial crystallization, microcrystals were found in several conditions of JCSG - plus and Structure screen 1&2 (Molecular Dimensions Ltd, UK) commercial screens. Optimization procedures including protein and salt concentration variations resulted in appearance of crystals sufficient for x-ray diffraction experiments. Needle clusters and plate-like crystals with size about 1.3 × 0.059 mm and 0.2 × 0.065 mm, respectively, were obtained. The datasets were collected at resolution ranging from 1.8 to 2.4 Å.
This work was partially supported by the Ministry of Education, Youth and Sport of the Czech Republic – projects CENAKVA (No. CZ 1. 05/2.1. 00/01.0024) and CENAKVA II (No. LO1205 under NPU I program) and GAJU grant (134/2013/ Z 2014 FUUP), and by the GACR 207/12/0775.