at Google ScholarTitle data
Claren, Jörg ; Malisi, Christoph ; Höcker, Birte ; Sterner, Reinhard:
Establishing wild-type levels of catalytic activity on natural and artificial (βα)8-barrel protein scaffolds.
In: Proceedings of the National Academy of Sciences of the United States of America.
Vol. 106
(2009)
Issue 10
.
- pp. 3704-3709.
ISSN 1091-6490
DOI: https://doi.org/10.1073/pnas.0810342106
Abstract in another language
The generation of high levels of new catalytic activities on natural and artificial protein scaffolds is a major goal of enzyme engineering. Here, we used random mutagenesis and selection in vivo to establish a sugar isomerisation reaction on both a natural (beta alpha)(8)-barrel enzyme and a catalytically inert chimeric (beta alpha)(8)-barrel scaffold, which was generated by the recombination of 2 (beta alpha)(4)-half barrels. The best evolved variants show turnover numbers and substrate affinities that are similar to those of wild-type enzymes catalyzing the same reaction. The determination of the crystal structure of the most proficient variant allowed us to model the substrate sugar in the novel active site and to elucidate the mechanistic basis of the newly established activity. The results demonstrate that natural and inert artificial protein scaffolds can be converted into highly proficient enzymes in the laboratory, and provide insights into the mechanisms of enzyme evolution.
Further data
| Item Type: | Article in a journal |
|---|---|
| Refereed: | Yes |
| Institutions of the University: | Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry > Chair Biochemistry - Univ.-Prof. Dr. Birte Höcker Faculties Faculties > Faculty of Biology, Chemistry and Earth Sciences Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry |
| Result of work at the UBT: | No |
| DDC Subjects: | 500 Science > 500 Natural sciences 500 Science > 570 Life sciences, biology |
| Date Deposited: | 27 Jan 2021 07:13 |
| Last Modified: | 05 Sep 2022 07:42 |
| URI: | https://eref.uni-bayreuth.de/id/eprint/62428 |