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Balancing the Stability–Activity Trade-Off by Fine-Tuning Dehalogenase Access Tunnels
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SYSNO ASEP 0452900 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Balancing the Stability–Activity Trade-Off by Fine-Tuning Dehalogenase Access Tunnels Tvůrce(i) 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)Zdroj.dok. ChemCatChem. - : Wiley - ISSN 1867-3880
Roč. 7, č. 4 (2015), s. 648-659Poč.str. 12 s. Jazyk dok. eng - angličtina Země vyd. DE - Německo Klíč. slova alkanes ; enzymes ; catalysis ; halogenation ; molecular dynamics ; protein engineering Vědní obor RIV CE - Biochemie Institucionální podpora RVO:67179843 - RVO:67179843 UT WOS 000349917900016 DOI 10.1002/cctc.201402792 Anotace 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. Pracoviště Ústav výzkumu globální změny Kontakt Nikola Šviková, svikova.n@czechglobe.cz, Tel.: 511 192 268 Rok sběru 2016
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