Document Detail


Use of "small but smart" libraries to enhance the enantioselectivity of an esterase from Bacillus stearothermophilus towards tetrahydrofuran-3-yl acetate.
MedLine Citation:
PMID:  23331978     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
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.
Authors:
Alberto Nobili; Markus G Gall; Ioannis V Pavlidis; Mark L Thompson; Marlen Schmidt; Uwe T Bornscheuer
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-1-19
Journal Detail:
Title:  The FEBS journal     Volume:  -     ISSN:  1742-4658     ISO Abbreviation:  FEBS J.     Publication Date:  2013 Jan 
Date Detail:
Created Date:  2013-1-21     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101229646     Medline TA:  FEBS J     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
© 2013 The Authors Journal compilation © 2013 FEBS.
Affiliation:
Dept. of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, D-17487, Greifswald, Germany.
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