Publication details

Balancing the Stability-Activity Trade-off by Fine-Tuning Dehalogenase Access Tunnels.

Authors

LIŠKOVÁ Veronika BEDNÁŘ David PRUDNIKOVA T. REZACOVA P. KOUDELÁKOVÁ Táňa ŠEBESTOVÁ Eva KUTA- SMATANOVÁ I., BREZOVSKÝ Jan CHALOUPKOVÁ Radka DAMBORSKÝ Jiří

Year of publication 2015
Type Article in Periodical
Magazine / Source ChemCatChem
MU Faculty or unit

Faculty of Science

Citation
Web http://loschmidt.chemi.muni.cz/peg/wp-content/uploads/2015/03/chemcatchem15.pdf
Doi http://dx.doi.org/10.1002/cctc.201402792
Field Biochemistry
Keywords haloalkane dehalogenase DhaA;access tunnel
Description 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 stabilized enzyme with superior catalytic activity, two of the four originally modified residues were randomized. The resulting mutant F176G exhibited 10- and 32-times enhanced activity towards 1,2-dibromoethane in buffer and 40% (v/v) DMSO, respectively, while retaining high stability. Structural and molecular dynamics analyses showed that the new variant exhibited superior activity because the F176G mutation increased the radius of the tunnel’s mouth and the mobility of alpha-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 four-point variant. Delicate balance between activity and stability of enzymes can be manipulated by fine-tuning the diameter and dynamics of their access tunnels
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