Deciphering the Structural Basis of High Thermostability of Dehalogenase from Psychrophilic Bacterium Marinobacter sp. ELB17

0511267 - ÚOCHB 2020 RIV CH eng J - Článek v odborném periodiku
Chrást, L. - Tratsiak, Katsiaryna - Planas-Iglesias, J. - Daniel, L. - Prudnikova, T. - Brezovský, J. - Bednář, D. - Kutá Smatanová, I. - Chaloupková, R. - Damborský, J.
Deciphering the Structural Basis of High Thermostability of Dehalogenase from Psychrophilic Bacterium Marinobacter sp. ELB17.
Microorganisms. Roč. 7, č. 11 (2019), č. článku 498. E-ISSN 2076-2607
Grant CEP: GA MŠMT(CZ) LM2015047
Institucionální podpora: RVO:61388963
Klíčová slova: haloalkane dehalogenase * thermostability * psychrophile * access tunnel * dimer * catalytic pentad * enantiselectivity
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 4.152, rok: 2019
Způsob publikování: Open access
https://www.mdpi.com/2076-2607/7/11/498

Haloalkane dehalogenases are enzymes with a broad application potential in biocatalysis, bioremediation, biosensing and cell imaging. The new haloalkane dehalogenase DmxA originating from the psychrophilic bacterium Marinobacter sp. ELB17 surprisingly possesses the highest thermal stability (apparent melting temperature Tm,app = 65.9 °C) of all biochemically characterized wild type haloalkane dehalogenases belonging to subfamily II. The enzyme was successfully expressed and its crystal structure was solved at 1.45 Å resolution. DmxA structure contains several features distinct from known members of haloalkane dehalogenase family: (i) a unique composition of catalytic residues, (ii) a dimeric state mediated by a disulfide bridge, and (iii) narrow tunnels connecting the enzyme active site with the surrounding solvent. The importance of narrow tunnels in such paradoxically high stability of DmxA enzyme was confirmed by computational protein design and mutagenesis experiments.
Trvalý link: http://hdl.handle.net/11104/0301587