Construction and characterisation of a genetically engineered Escherichia coli strain for the epoxide hydrolase-catalysed kinetic resolution of epoxides
| Autor: | H. Visser, M. de Oliveira Vil Filho, Andreas Liese, Jan C. Verdoes, C.A.G.M. Weijers |
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| Jazyk: | angličtina |
| Rok vydání: | 2003 |
| Předmět: |
Epoxide
Rhodotorula medicine.disease_cause Biochemistry Microbiology Catalysis Inclusion bodies Kinetic resolution chemistry.chemical_compound encoding gene Microbiologie medicine Epoxide hydrolase Escherichia coli rhodotorula-glutinis VLAG degradation biology Chemistry Organic Chemistry Substrate (chemistry) abnormal protein enantioselective hydrolysis biology.organism_classification Enterobacteriaceae groel Organische Chemie tools trigger factor Wageningen University Biotechnology |
| Zdroj: | Biocatalysis and Biotransformation 21 (2003) 1 Biocatalysis and Biotransformation, 21(1), 33-40 |
| ISSN: | 1024-2422 |
| Popis: | The Rhodotorula glutinis epoxide hydrolase, Eph1, was produced in the heterologous host Escherichia coli BL21(DE3) in order to develop a highly effective epoxide hydrolysis system. A 138-fold increase in Eph1 activity was found in cell extracts of the recombinant E. coli when compared to cell extracts of Rhodotorula glutinis, despite the formation of Eph1 inclusion bodies. Optimization of cultivation conditions and co-expression of molecular chaperones resulted in a further increase in activity and a reduction of the inclusion bodies formation, respectively. Compared to Rhodotorula glutinis cells and cell extracts, a total increase in Eph1 activity of over 200 times was found for both Escherichia coli cells and crude enzyme preparations of these cells. The improved conditions for recombinant Eph1 production were used to demonstrate the Eph1-catalysed kinetic resolution of a new Eph1 substrate, 1-oxaspiro[2.5]octane-2-carbonitrile. |
| Databáze: | OpenAIRE |
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