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

J 2015

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

LIŠKOVÁ, Veronika, David BEDNÁŘ, T. PRUDNIKOVA, P. REZACOVA, Táňa KOUDELÁKOVÁ et. al.

Basic information

Original name

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

Authors

LIŠKOVÁ, Veronika (203 Czech Republic, belonging to the institution), David BEDNÁŘ (203 Czech Republic, belonging to the institution), T. PRUDNIKOVA (112 Belarus), P. REZACOVA (203 Czech Republic), Táňa KOUDELÁKOVÁ (203 Czech Republic, belonging to the institution), Eva ŠEBESTOVÁ (203 Czech Republic, belonging to the institution), I., KUTA- SMATANOVÁ (203 Czech Republic), Jan BREZOVSKÝ (203 Czech Republic, belonging to the institution), Radka CHALOUPKOVÁ (203 Czech Republic, belonging to the institution) and Jiří DAMBORSKÝ (203 Czech Republic, guarantor, belonging to the institution)

Edition

ChemCatChem, 2015, 1867-3880

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 4.724

RIV identification code

RIV/00216224:14310/15:00080815

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1002/cctc.201402792

UT WoS

000349917900016

Keywords in English

haloalkane dehalogenase DhaA;access tunnel

Abstract

V originále

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

Links

EE2.3.30.0037, research and development project

Name: Zaměstnáním nejlepších mladých vědců k rozvoji mezinárodní spolupráce

GAP207/12/0775, research and development project

Name: Strukturně-funkční vztahy haloalkan dehalogenas

Investor: Czech Science Foundation

LH14027, research and development project

Name: Nové koncepty a nástroje pro racionální design enzymů

Investor: Ministry of Education, Youth and Sports of the CR

LM2010005, research and development project

Name: Velká infrastruktura CESNET (Acronym: VI CESNET)

Investor: Ministry of Education, Youth and Sports of the CR

LO1214, research and development project

Name: Centrum pro výzkum toxických látek v prostředí (Acronym: RECETOX)

Investor: Ministry of Education, Youth and Sports of the CR

Citovat

LIŠKOVÁ, Veronika, David BEDNÁŘ, T. PRUDNIKOVA, P. REZACOVA, Táňa KOUDELÁKOVÁ, Eva ŠEBESTOVÁ, I., KUTA- SMATANOVÁ, Jan BREZOVSKÝ, Radka CHALOUPKOVÁ and Jiří DAMBORSKÝ. Balancing the Stability-Activity Trade-off by Fine-Tuning Dehalogenase Access Tunnels. ChemCatChem. 2015, vol. 7, No 4, p. 648-659. ISSN 1867-3880. Available from: https://dx.doi.org/10.1002/cctc.201402792.

@article{1300087,
   author = {Lišková, Veronika and Bednář, David and Prudnikova, T. and Rezacova, P. and Koudeláková, Táňa and Šebestová, Eva and Kutaand Smatanová, I., and Brezovský, Jan and Chaloupková, Radka and Damborský, Jiří},
   article_number = {4},
   doi = {http://dx.doi.org/10.1002/cctc.201402792},
   keywords = {haloalkane dehalogenase DhaA;access tunnel},
   language = {eng},
   issn = {1867-3880},
   journal = {ChemCatChem},
   title = {Balancing the Stability-Activity Trade-off by Fine-Tuning Dehalogenase Access Tunnels.},
   url = {http://loschmidt.chemi.muni.cz/peg/wp-content/uploads/2015/03/chemcatchem15.pdf},
   volume = {7},
   year = {2015}
}

TY  - JOUR
ID  - 1300087
AU  - Lišková, Veronika - Bednář, David - Prudnikova, T. - Rezacova, P. - Koudeláková, Táňa - Šebestová, Eva - Kuta- Smatanová, I., - Brezovský, Jan - Chaloupková, Radka - Damborský, Jiří
PY  - 2015
TI  - Balancing the Stability-Activity Trade-off by Fine-Tuning Dehalogenase Access Tunnels.
JF  - ChemCatChem
VL  - 7
IS  - 4
SP  - 648-659
EP  - 648-659
SN  - 18673880
KW  - haloalkane dehalogenase DhaA;access tunnel
UR  - http://loschmidt.chemi.muni.cz/peg/wp-content/uploads/2015/03/chemcatchem15.pdf
L2  - http://loschmidt.chemi.muni.cz/peg/wp-content/uploads/2015/03/chemcatchem15.pdf
N2  - 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
ER  -

LIŠKOVÁ, Veronika, David BEDNÁŘ, T. PRUDNIKOVA, P. REZACOVA, Táňa KOUDELÁKOVÁ, Eva ŠEBESTOVÁ, I., KUTA- SMATANOVÁ, Jan BREZOVSKÝ, Radka CHALOUPKOVÁ and Jiří DAMBORSKÝ. Balancing the Stability-Activity Trade-off by Fine-Tuning Dehalogenase Access Tunnels. \textit{ChemCatChem}. 2015, vol.~7, No~4, p.~648-659. ISSN~1867-3880. Available from: https://dx.doi.org/10.1002/cctc.201402792.

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