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Crystallographic analysis of 1,2,3-trichloropropane biodegradation by the haloalkane dehalogenase DhaA31
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SYSNO ASEP 0435285 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Crystallographic analysis of 1,2,3-trichloropropane biodegradation by the haloalkane dehalogenase DhaA31 Author(s) Lahoda, M. (CZ)
Mesters, J. R. (DE)
Stsiapanava, A. (CZ)
Chaloupková, R. (CZ)
Kutý, Michal (UEK-B)
Damborský, J. (CZ)
Kutá-Smatanová, Ivana (UEK-B) RIDSource Title Acta Crystallographica Section D-Biological Crystallography. - : WILEY-BLACKWELL - ISSN 0907-4449
Roč. 70, FEB 2014 (2014), s. 209-217Number of pages 9 s. Language eng - English Country DK - Denmark Keywords DhaA31 ; substrate-free ; 3rk4 ; complex with TCP ; 4fwb ; wild-type DhaA ; 4hzg Subject RIV CE - Biochemistry Institutional support RVO:67179843 - RVO:67179843 UT WOS 000331554500001 DOI https://doi.org/10.1107/S1399004713026254 Annotation Haloalkane dehalogenases catalyze the hydrolytic cleavage of carbon-halogen bonds, which is a key step in the aerobic mineralization of many environmental pollutants. One important pollutant is the toxic and anthropogenic compound 1,2,3-trichloropropane (TCP). Rational design was combined with saturation mutagenesis to obtain the haloalkane dehalogenase variant DhaA31, which displays an increased catalytic activity towards TCP. Here, the 1.31 angstrom resolution crystal structure of substrate-free DhaA31, the 1.26 angstrom resolution structure of DhaA31 in complex with TCP and the 1.95 angstrom resolution structure of wild-type DhaA are reported. Crystals of the enzyme-substrate complex were successfully obtained by adding volatile TCP to the reservoir after crystallization at pH 6.5 and room temperature. Comparison of the substrate-free structure with that of the DhaA31 enzyme-substrate complex reveals that the nucleophilic Asp106 changes its conformation from an inactive to an active state during the catalytic cycle. The positions of three chloride ions found inside the active site of the enzyme indicate a possible pathway for halide release from the active site through the main tunnel. Comparison of the DhaA31 variant with wild-type DhaA revealed that the introduced substitutions reduce the volume and the solvent-accessibility of the active-site pocket. Workplace Global Change Research Institute Contact Nikola Šviková, svikova.n@czechglobe.cz, Tel.: 511 192 268 Year of Publishing 2015
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