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A highly stereoselective recombinant alcohol dehydrogenase aus 'Pseudomonas fluorescens' DSM50106
- The alcohol dehydrogenase was biochemically characterized. A broad range of arylaliphatic ketones is efficiently reduced to the corresponding optically active (R)-alcohols by a recombinant alcohol dehydrogenase (PF-ADH) produced by overexpression in 'Escherichia coli'. PF-ADH shows high activity and stereoselectivity in the reduction of acetophenone and various derivatives (45-99%), as well as in the reduction of 3-oxy-butyric acid methyl ester and 3-oxy-butyric acid methyl ester and 3-oxy-hexanoic acid ethyl ester (>99%). The highest activity was observed between 10 and 20°C. The copfactor NADH can be efficiently recycled by the addition of 10-20% of iso-propanol. A flow-through-polarimetry-based assay to determine oxidoreductase activity and stereoselectivity is described.
| Author: | Petra Hildebrandt |
|---|---|
| URN: | urn:nbn:de:gbv:9-000016-5 |
| Title Additional (German): | Eine hochgradig stereoselektive rekombinante Alkoholdehydrogenase aus 'Pseudomonas fluorescens' DSM50106 |
| Advisor: | Prof. Dr. Uwe T. Bornscheuer |
| Document Type: | Doctoral Thesis |
| Language: | English |
| Date of Publication (online): | 2006/07/03 |
| Granting Institution: | Ernst-Moritz-Arndt-Universität, Mathematisch-Naturwissenschaftliche Fakultät (bis 31.05.2018) |
| Date of final exam: | 2005/11/26 |
| Release Date: | 2006/07/03 |
| Tag: | Ketoreductase; Oxidoreductase; alcohol dehydrogenase; enantioselectivity; recombinant enzyme |
| GND Keyword: | Oxocarbonsäureester |
| Faculties: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie |
| DDC class: | 500 Naturwissenschaften und Mathematik / 540 Chemie |