Title : Directed evolution of an esterase from Pseudomonas fluorescens. Random mutagenesis by error-prone PCR or a mutator strain and identification of mutants showing enhanced enantioselectivity by a resorufin-based fluorescence assay - Henke_1999_Biol.Chem_380_1029 |
Author(s) : Henke E , Bornscheuer UT |
Ref : Biol Chem , 380 :1029 , 1999 |
Abstract :
The gene encoding an esterase from Pseudomonas fluorescens (PFE) was subjected to random mutagenesis by error-prone PCR or by using the mutator strain Epicurian coli XL1-Red. Enzyme libraries were then created in microtiter plates by expression of PFE-variants in E. coli. These were assayed for improved enantioselectivity in a Beckman robot system using optically pure (R)- or (S)-3-phenylbutyric acid resorufin esters, resulting in the identification of several mutants showing up to almost two-fold enantioselectivity (E(true) = 5.2 to 6.6) compared to wild-type PFE (E(true) = 3.5). |
PubMedSearch : Henke_1999_Biol.Chem_380_1029 |
PubMedID: 10494857 |
Gene_locus related to this paper: psefl-este |
Substrate | Resorufin-butyrate |
Gene_locus | psefl-este |
Henke E, Bornscheuer UT (1999)
Directed evolution of an esterase from Pseudomonas fluorescens. Random mutagenesis by error-prone PCR or a mutator strain and identification of mutants showing enhanced enantioselectivity by a resorufin-based fluorescence assay
Biol Chem
380 :1029
Henke E, Bornscheuer UT (1999)
Biol Chem
380 :1029