Balancing the stability-activity trade-off by fine-tuning dehalogenase access tunnels
Authors:
- Veronika Liskova,
- David Bednar,
- Tatyana Prudnikova,
- Pavlina Rezacova,
- Tana Koudelakova,
- Eva Sebestova,
- Ivana Kuta Smatanova,
- Jan Brezovsky,
- Radka Chaloupkova,
- Jiri Damborsky
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 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 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. Tuning enzyme access tunnels: The catalytic activity and stability of enzymes can be inversely related. It is shown that haloalkane dehalogenases with high stability and activity can be created by fine-tuning the diameter and dynamics of the access tunnels that connect buried active sites to the surrounding bulk solvent.
- Record ID
- UAMb4b640549a824f8e92185386e66bd378
- Author
- Journal series
- Chemcatchem, ISSN 1867-3880
- Issue year
- 2015
- Vol
- 7
- Pages
- 648-659
- ASJC Classification
- ; ; ;
- DOI
- DOI:10.1002/cctc.201402792 Opening in a new tab
- Language
- (en) English
- Score (nominal)
- 35
- Score source
- journalList
- Score
- Publication indicators
- = 31; = 33; : 2015 = 1.007; : 2015 (2 years) = 4.724 - 2015 (5 years) =5.107
- Citation count
- 44
Share
- Uniform Resource Identifier
- https://researchportal.amu.edu.pl/info/article/UAMb4b640549a824f8e92185386e66bd378/
- URN
urn:amu-prod:UAMb4b640549a824f8e92185386e66bd378
* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or PerishOpening in a new tab system.