TY - JOUR
T1 - Crystallization and crystallographic analysis of a bradyrhizobium elkanii usda94 haloalkane dehalogenase variant with an eliminated halide-binding site
AU - Prudnikova, Tatyana
AU - Kascakova, Barbora
AU - Mesters, Jeroen R.
AU - Grinkevich, Pavel
AU - Havlickova, Petra
AU - Mazur, Andrii
AU - Shaposhnikova, Anastasiia
AU - Chaloupkova, Radka
AU - Damborsky, Jiri
AU - Kuty, Michal
AU - Kuta Smatanova, Ivana
N1 - Funding Information:
Funding: This work was supported by the Grant Agency of the Czech Republic 17-24321S; DAAD mobility grant DAAD-16-09; ERDF project CZ.02.1.01/0.0/0.0/15_003/0000441; Ministry of Education, Youth and Sports of the Czech Republic (CZ.1.05/2.1.00/01.0024, CZ.1.05/2.1.00/01.0001, and LM2015055); GAJU 17/2019/P.
Publisher Copyright:
© 2019 by the authors. Licensee MDPI, Basel, Switzerland.
Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2019/7
Y1 - 2019/7
N2 - Haloalkane dehalogenases are a very important class of microbial enzymes for environmental detoxification of halogenated pollutants, for biocatalysis, biosensing and molecular tagging. The double mutant (Ile44Leu + Gln102His) of the haloalkane dehalogenase DbeA from Bradyrhizobium elkanii USDA94 (DbeA∆Cl) was constructed to study the role of the second halide-binding site previously discovered in the wild-type structure. The variant is less active, less stable in the presence of chloride ions and exhibits significantly altered substrate specificity when compared with the DbeAwt. DbeA∆Cl was crystallized using the sitting-drop vapour-diffusion procedure with further optimization by the random microseeding technique. The crystal structure of the DbeA∆Cl has been determined and refined to the 1.4 Å resolution. The DbeA∆Cl crystals belong to monoclinic space group C121. The DbeA∆Cl molecular structure was characterized and compared with five known haloalkane dehalogenases selected from the Protein Data Bank.
AB - Haloalkane dehalogenases are a very important class of microbial enzymes for environmental detoxification of halogenated pollutants, for biocatalysis, biosensing and molecular tagging. The double mutant (Ile44Leu + Gln102His) of the haloalkane dehalogenase DbeA from Bradyrhizobium elkanii USDA94 (DbeA∆Cl) was constructed to study the role of the second halide-binding site previously discovered in the wild-type structure. The variant is less active, less stable in the presence of chloride ions and exhibits significantly altered substrate specificity when compared with the DbeAwt. DbeA∆Cl was crystallized using the sitting-drop vapour-diffusion procedure with further optimization by the random microseeding technique. The crystal structure of the DbeA∆Cl has been determined and refined to the 1.4 Å resolution. The DbeA∆Cl crystals belong to monoclinic space group C121. The DbeA∆Cl molecular structure was characterized and compared with five known haloalkane dehalogenases selected from the Protein Data Bank.
UR - http://www.scopus.com/inward/record.url?scp=85071098152&partnerID=8YFLogxK
U2 - 10.3390/cryst9070375
DO - 10.3390/cryst9070375
M3 - Journal articles
AN - SCOPUS:85071098152
SN - 2073-4352
VL - 9
JO - Crystals
JF - Crystals
IS - 7
M1 - 375
ER -
