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Directed Evolution of the Epoxide Hydrolase from Aspergillus niger
An efficient and genetically stable expression system for the directed evolution of epoxide hydrolase from Aspergillus niger (ANEH) has been constructed. Error prone polymerase chain reaction (PCR) with defined mutation rates was used to create biodiversity in two libraries of mutants. Screening for...
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| Published in: | Biocatalysis and biotransformation 2003-12, Vol.21 (6), p.357-364 |
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| Main Authors: | , , , , , , , , , |
| Format: | Article |
| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c450t-b9352e49bad900c13ad5ba48d210caf2a8e50bb223edaf77d67fe22a891f69833 |
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| cites | cdi_FETCH-LOGICAL-c450t-b9352e49bad900c13ad5ba48d210caf2a8e50bb223edaf77d67fe22a891f69833 |
| container_end_page | 364 |
| container_issue | 6 |
| container_start_page | 357 |
| container_title | Biocatalysis and biotransformation |
| container_volume | 21 |
| creator | Cedrone, Frederic Niel, Sebastien Roca, Sanja Bhatnagar, Tej Ait-abdelkader, Nadra Torre, Claudia Krumm, Holger Maichele, Andrea T. Reetz, Manfred C. Baratti, Jacques |
| description | An efficient and genetically stable expression system for the directed evolution of epoxide hydrolase from Aspergillus niger (ANEH) has been constructed. Error prone polymerase chain reaction (PCR) with defined mutation rates was used to create biodiversity in two libraries of mutants. Screening for activity allowed the isolation of clones with improved properties. One of these clones shows an expression level 3.4 higher than the original wild type clone in E. coli SG13009 and a 3.3 fold increased catalytic efficiency on 4-(p-nitrophenoxy)-1,2-epoxybutane. In addition, a screening assay for determining the enantioselectivity in the kinetic resolution of styrene oxide has been established using mass spectrometry. |
| doi_str_mv | 10.1080/102420310001630137 |
| format | article |
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Reetz, Manfred</creatorcontrib><creatorcontrib>C. Baratti, Jacques</creatorcontrib><title>Directed Evolution of the Epoxide Hydrolase from Aspergillus niger</title><title>Biocatalysis and biotransformation</title><description>An efficient and genetically stable expression system for the directed evolution of epoxide hydrolase from Aspergillus niger (ANEH) has been constructed. Error prone polymerase chain reaction (PCR) with defined mutation rates was used to create biodiversity in two libraries of mutants. Screening for activity allowed the isolation of clones with improved properties. One of these clones shows an expression level 3.4 higher than the original wild type clone in E. coli SG13009 and a 3.3 fold increased catalytic efficiency on 4-(p-nitrophenoxy)-1,2-epoxybutane. In addition, a screening assay for determining the enantioselectivity in the kinetic resolution of styrene oxide has been established using mass spectrometry.</description><subject>Aspergillus niger</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Catalytic efficiency</subject><subject>Directed evolution</subject><subject>Enantioselectivity</subject><subject>Enzyme engineering</subject><subject>Epoxide hydrolase</subject><subject>Expression system</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Methods. Procedures. 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| fulltext | fulltext |
| identifier | ISSN: 1024-2422 |
| ispartof | Biocatalysis and biotransformation, 2003-12, Vol.21 (6), p.357-364 |
| issn | 1024-2422 1029-2446 |
| language | eng |
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| source | Taylor and Francis Science and Technology Collection |
| subjects | Aspergillus niger Biological and medical sciences Biotechnology Catalytic efficiency Directed evolution Enantioselectivity Enzyme engineering Epoxide hydrolase Expression system Fundamental and applied biological sciences. Psychology Methods. Procedures. Technologies Protein engineering |
| title | Directed Evolution of the Epoxide Hydrolase from Aspergillus niger |
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