Gerichtete Evolution als Methode zur Erzeugung enantioselektiver Cyclohexanonmonooxygenasen (CHMOs) für die Katalyse von Baeyer-Villiger-Reaktionen
- Aim of the project was to improve the enantioselectivity of Cyclohexanone Monooxygenase (CHMO) towards 4 hydroxycyclohexanone in the Baeyer-Villiger reaction using the principle of directed evolution. Mutants were produced by epPCR and saturation mutagenesis. Reaction conditions were optimized and a medium-throughput gas chromatography-based screening system for the identification of improved mutants was developed, so that 800 mutants per day can be screened. After two rounds of directed evolution the enantioselectivity of CHMO towards the substrate was increased from 9 to 90%. A mutant of reverse selectivity of 79% was found which was also able to catalyse on other substrates with excellent enantioselectivities (up to 99% ee).
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| Author: | Toni SchneiderGND |
|---|---|
| URN: | urn:nbn:de:hbz:294-12354 |
| Referee: | Manfred T. ReetzGND, Martin FeigelGND |
| Document Type: | Doctoral Thesis |
| Language: | German |
| Date of Publication (online): | 2005/02/18 |
| Date of first Publication: | 2005/02/18 |
| Publishing Institution: | Ruhr-Universität Bochum, Universitätsbibliothek |
| Granting Institution: | Ruhr-Universität Bochum, Fakultät für Chemie und Biochemie |
| Date of final exam: | 2005/01/20 |
| Creating Corporation: | Fakultät für Chemie und Biochemie |
| GND-Keyword: | Evaluation; Mutagenese; Screening; Kapillargaschromatographie; Asymmetrische Synthese |
| Institutes/Facilities: | Max-Planck-Institut für Kohlenforschung, Mülheim an der Ruhr |
| Dewey Decimal Classification: | Naturwissenschaften und Mathematik / Chemie, Kristallographie, Mineralogie |
| faculties: | Fakultät für Chemie und Biochemie |
| Licence (German): |  Keine Creative Commons Lizenz - es gelten der Veröffentlichungsvertrag und das deutsche Urheberrecht |
