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| rdf:type
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| Description
| - A large number of chemical reactions are catalyzed in the active sites localized on the protein surface, but many enzymes have their active site buried and connected with the surrounding solvent by an access tunnel. The aim of this study was to identify structural features of the enzymes possessing buried active sites governing their stability and resistance to organic cosolvents. The haloalkane dehalogenase DhaA from Rhodococcus rhodochrous NCIMB 13064 (EC 3.8.1.5) has been chosen as a model system for this protein engineering study. Here, we constructed a systematic set of DhaA variants by random and site-directed mutagenesis, and characterized them structurally, kinetically, and computationally. The acquired complementary data provided detailed insights into the structure–stability relationships of this enzyme and enabled us to construct a biocatalyst with substantially improved stability and resistance to organic cosolvent. We conclude that saturation mutagenesis of the residues lining the access tunnels is a widely applicable strategy for engineering resistance to organic cosolvents and elevated temperatures in the proteins with buried active sites.
- A large number of chemical reactions are catalyzed in the active sites localized on the protein surface, but many enzymes have their active site buried and connected with the surrounding solvent by an access tunnel. The aim of this study was to identify structural features of the enzymes possessing buried active sites governing their stability and resistance to organic cosolvents. The haloalkane dehalogenase DhaA from Rhodococcus rhodochrous NCIMB 13064 (EC 3.8.1.5) has been chosen as a model system for this protein engineering study. Here, we constructed a systematic set of DhaA variants by random and site-directed mutagenesis, and characterized them structurally, kinetically, and computationally. The acquired complementary data provided detailed insights into the structure–stability relationships of this enzyme and enabled us to construct a biocatalyst with substantially improved stability and resistance to organic cosolvent. We conclude that saturation mutagenesis of the residues lining the access tunnels is a widely applicable strategy for engineering resistance to organic cosolvents and elevated temperatures in the proteins with buried active sites. (en)
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| Title
| - Engineering Enzyme Stability and Resistance to an Organic Cosolvent by Modification of Residues in the Access Tunnel
- Engineering Enzyme Stability and Resistance to an Organic Cosolvent by Modification of Residues in the Access Tunnel (en)
|
| skos:prefLabel
| - Engineering Enzyme Stability and Resistance to an Organic Cosolvent by Modification of Residues in the Access Tunnel
- Engineering Enzyme Stability and Resistance to an Organic Cosolvent by Modification of Residues in the Access Tunnel (en)
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| skos:notation
| - RIV/68378050:_____/13:00425304!RIV14-AV0-68378050
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| http://linked.open...avai/predkladatel
| |
| http://linked.open...avai/riv/aktivita
| |
| http://linked.open...avai/riv/aktivity
| - I, P(ED0001/01/01), P(GA203/08/0114), P(GAP202/10/1435), P(GAP207/12/0775), P(LC06010), Z(MSM0021622412)
|
| http://linked.open...iv/cisloPeriodika
| |
| http://linked.open...vai/riv/dodaniDat
| |
| http://linked.open...aciTvurceVysledku
| |
| http://linked.open.../riv/druhVysledku
| |
| http://linked.open...iv/duvernostUdaju
| |
| http://linked.open...titaPredkladatele
| |
| http://linked.open...dnocenehoVysledku
| |
| http://linked.open...ai/riv/idVysledku
| - RIV/68378050:_____/13:00425304
|
| http://linked.open...riv/jazykVysledku
| |
| http://linked.open.../riv/klicovaSlova
| - directed evolution; enzyme catalysis; enzymes; protein engineering; protein stability (en)
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| http://linked.open.../riv/klicoveSlovo
| |
| http://linked.open...odStatuVydavatele
| - DE - Spolková republika Německo
|
| http://linked.open...ontrolniKodProRIV
| |
| http://linked.open...i/riv/nazevZdroje
| - Angewandte Chemie. International Edition in English
|
| http://linked.open...in/vavai/riv/obor
| |
| http://linked.open...ichTvurcuVysledku
| |
| http://linked.open...cetTvurcuVysledku
| |
| http://linked.open...vavai/riv/projekt
| |
| http://linked.open...UplatneniVysledku
| |
| http://linked.open...v/svazekPeriodika
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| http://linked.open...iv/tvurceVysledku
| - Chaloupková, R.
- Damborský, J.
- Ettrich, Rüdiger
- Kutá-Smatanová, Ivana
- Khabiri, Morteza
- Řezáčová, Pavlína
- Prokop, Z.
- Bornscheuer, U.T.
- Březovský, J.
- Hesseler, M.
- Koudeláková, T.
- Kulik, D.
- Plevaka, M.
- Šebestová, E.
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| http://linked.open...ain/vavai/riv/wos
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| http://linked.open...n/vavai/riv/zamer
| |
| issn
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| number of pages
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| http://bibframe.org/vocab/doi
| |
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