Paper Report for: Jochens_2010_Protein.Eng.Des.Sel_23_903
Reference
Title: Thermostabilization of an esterase by alignment-guided focussed directed evolution Jochens H, Aerts D, Bornscheuer UT Ref: Protein Engineering Des Sel, 23:903, 2010 : PubMed
Site-saturation libraries of the Pseudomonas fluorescens esterase were created targeting three surface positions to increase its thermostability on the basis of the B-factor iterative test principle. All three positions were saturated simultaneously using our recently developed protocol for the design of 'small, but smart' mutant libraries bearing only consensus-like mutations. Hence, the library size could be significantly reduced while ensuring a high hit rate. Variants could be identified that showed significantly improved stability (8 degrees C higher compared with the wild type) without compromising specific activity. Subsequent iterative saturation mutagenesis gave an esterase mutant with a 9 degrees C increased melting point, but unchanged catalytic properties.
        
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Jochens H, Aerts D, Bornscheuer UT (2010) Thermostabilization of an esterase by alignment-guided focussed directed evolution Protein Engineering Des Sel23: 903-9
Jochens H, Aerts D, Bornscheuer UT (2010) Protein Engineering Des Sel23: 903-9