Publication details

Engineering Enzyme Stability and Resistance to an Organic Cosolvent by Modification of Residues in the Access Tunnel

Authors

KOUDELÁKOVÁ Táňa CHALOUPKOVÁ Radka BREZOVSKÝ Jan PROKOP Zbyněk ŠEBESTOVÁ Eva HESSELER M. KHABIRI M. PLEVAKA M. KULIK D. KUTA SMATANOVA I. REZACOVA P. ETTRICH R. BORNSCHEUER U.T. DAMBORSKÝ Jiří

Type Article in Periodical
Magazine / Source Angewandte Chemie International Edition
MU Faculty or unit

Faculty of Science

Citation
Doi http://dx.doi.org/10.1002/anie.201206708
Field Biochemistry
Keywords haloalkan dehalogenases
Description Mutations targeting as few as four residues lining the access tunnel extended enzyme’s half-life in 40% dimethyl sulfoxide from minutes to weeks (4,000-fold) and increased its melting temperature by 19 Grades C. Protein crystallography and molecular dynamics revealed that the tunnel residue packing is a key determinant of protein stability and the active-site accessibility for co-solvent molecules (red dots). The broad applicability of this concept was verified by analyzing twenty six proteins with buried active sites from all six enzyme classes.
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