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- Title
Balancing the Stability-Activity Trade-Off by Fine-Tuning Dehalogenase Access Tunnels.
- Authors
Liskova, Veronika; Bednar, David; Prudnikova, Tatyana; Rezacova, Pavlina; Koudelakova, Tana; Sebestova, Eva; Smatanova, Ivana Kuta; Brezovsky, Jan; Chaloupkova, Radka; Damborsky, Jiri
- Abstract
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 F 176 G 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 F 176 G mutation increased the radius of the tunnel's mouth and the mobility of α-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.
- Subjects
ENZYME stability; DEHALOGENASES; HALOALKANES; ORGANIC solvents; CATALYTIC activity; ETHYLENE dibromide
- Publication
ChemCatChem, 2015, Vol 7, Issue 4, p648
- ISSN
1867-3880
- Publication type
Article
- DOI
10.1002/cctc.201402792