| Use of "small but smart" libraries to enhance the enantioselectivity of an esterase from Bacillus stearothermophilus towards tetrahydrofuran-3-yl acetate. | |
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MedLine Citation:
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PMID: 23331978 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Two libraries of simultaneous double mutations in the active-site region of an esterase from Bacillus stearothermophilus (BsteE) were constructed to improve the enantioselectivity in the hydrolysis of tetrahydrofuran-3-yl acetate. As screening of large mutant libraries is hampered by the necessity of a GC/MS analysis, mutant libraries were designed using a "small, but smart" concept. The design of focused libraries was based on data derived from a structural alignment of 3,317 amino acid sequences of α/β hydrolase fold enzymes using the bioinformatic tool 3DM. In this way, the number of mutants to be screened was substantially reduced compared to a standard site-saturation mutagenesis approach. Whilst the wild type esterase exhibited only poor enantioselectivity (E = 4.3) in the hydrolysis of (S)-tetrahydrofuran-3-yl acetate the best variants obtained by this approach showed increased E-values of up to 10.4. Furthermore, some variants with inverted enantiopreference were found. © 2013 The Authors Journal compilation © 2013 FEBS. |
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Authors:
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Alberto Nobili; Markus G Gall; Ioannis V Pavlidis; Mark L Thompson; Marlen Schmidt; Uwe T Bornscheuer |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2013-1-19 |
Journal Detail:
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Title: The FEBS journal Volume: - ISSN: 1742-4658 ISO Abbreviation: FEBS J. Publication Date: 2013 Jan |
Date Detail:
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Created Date: 2013-1-21 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101229646 Medline TA: FEBS J Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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© 2013 The Authors Journal compilation © 2013 FEBS. |
Affiliation:
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Dept. of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, D-17487, Greifswald, Germany. |
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