Informace o publikaci

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

Autoři

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

Rok publikování 2015
Druh Článek v odborném periodiku
Časopis / Zdroj ChemCatChem
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
www http://loschmidt.chemi.muni.cz/peg/wp-content/uploads/2015/03/chemcatchem15.pdf
Doi http://dx.doi.org/10.1002/cctc.201402792
Obor Biochemie
Klíčová slova haloalkane dehalogenase DhaA;access tunnel
Popis 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
Související projekty:

Používáte starou verzi internetového prohlížeče. Doporučujeme aktualizovat Váš prohlížeč na nejnovější verzi.

Další info