Crystallization and Crystallographic Analysis of a Bradyrhizobium Elkanii USDA94 Haloalkane Dehalogenase Variant with an Eliminated Halide-Binding Site

0508541 - MBÚ 2020 RIV CH eng J - Článek v odborném periodiku
Prudnikova, Tatyana - Kascaková, B. - Mesters, J. R. - Grinkevich, P. - Havlíčková, P. - Mazur, Andrii - Shaposhnikova, Anastasiia - Chaloupková, R. - Dambrovský, J. - Kutý, Michal - Kutá Smatanová, Ivana
Crystallization and Crystallographic Analysis of a Bradyrhizobium Elkanii USDA94 Haloalkane Dehalogenase Variant with an Eliminated Halide-Binding Site.
Crystals. Roč. 9, č. 7 (2019), č. článku 375. ISSN 2073-4352. E-ISSN 2073-4352
Grant CEP: GA MŠMT(CZ) LM2015055
Grant ostatní: AV ČR(CZ) DAAD-16-09
Program: Bilaterální spolupráce
Výzkumná infrastruktura: C4SYS - 90055
Institucionální podpora: RVO:61388971
Klíčová slova: Haloalkane dehalogenase * halide-binding site * random microseeding
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 2.404, rok: 2019
Způsob publikování: Open access
https://www.mdpi.com/2073-4352/9/7/375

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 Delta 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 Delta Cl was crystallized using the sitting-drop vapour-diffusion procedure with further optimization by the random microseeding technique. The crystal structure of the DbeA Delta Cl has been determined and refined to the 1.4 angstrom resolution. The DbeA Delta Cl crystals belong to monoclinic space group C121. The DbeA Delta Cl molecular structure was characterized and compared with five known haloalkane dehalogenases selected from the Protein Data Bank.
Trvalý link: http://hdl.handle.net/11104/0299416