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Balancing the Stability–Activity Trade-Off by Fine-Tuning Dehalogenase Access Tunnels
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SYSNO ASEP 0452900 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Balancing the Stability–Activity Trade-Off by Fine-Tuning Dehalogenase Access Tunnels Author(s) Lišková, V. (CZ)
Bednář, D. (CZ)
Prudnikova, Tatyana (UEK-B) RID, SAI, ORCID
Řezáčová, P. (CZ)
Koudeláková, T. (CZ)
Šebestová, E. (CZ)
Kutá-Smatanová, Ivana (UEK-B) RID
Březovský, J. (CZ)
Chaloupková, R. (CZ)
Damborský, J. (CZ)Source Title ChemCatChem. - : Wiley - ISSN 1867-3880
Roč. 7, č. 4 (2015), s. 648-659Number of pages 12 s. Language eng - English Country DE - Germany Keywords alkanes ; enzymes ; catalysis ; halogenation ; molecular dynamics ; protein engineering Subject RIV CE - Biochemistry Institutional support RVO:67179843 - RVO:67179843 UT WOS 000349917900016 DOI 10.1002/cctc.201402792 Annotation A variant of the haloalkane dehalogenase DhaA with greatly enhanced stability and tolerance of organic solvents but reduced activity was created by mutating four residues in the access tunnel. To create a stabilised enzyme with superior catalytic activity, two of the four originally modified residues were randomised. The resulting mutant F176G exhibited 32- and 10-times enhanced activity towards 1,2-dibromoethane in buffer and 40% DMSO, respectively, upon retaining high stability. Structural and molecular dynamics analyses demonstrated that the new variant exhibited superior activity because the F176G mutation increased the radius of the tunnel’s mouth and the mobility of a-helices lining the tunnel. The new variant’s tunnel was open in 48% of trajectories, compared to 58% for the wild-type, but only 0.02% for the original fourpoint variant. Delicate balance between activity and stability of enzymes can be manipulated by fine-tuning the diameter and dynamics of their access tunnels. Workplace Global Change Research Institute Contact Nikola Šviková, svikova.n@czechglobe.cz, Tel.: 511 192 268 Year of Publishing 2016
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