Directed Enzyme Evolution via Small and Effective Neutral Drift Libraries

Nat Methods. 2008 Nov;5(11):939-42. doi: 10.1038/nmeth.1262. Epub 2008 Oct 19.

Abstract

Small libraries for directed evolution can be obtained by neutral drifts that maintain the protein's original function, yielding highly polymorphic, stable and evolvable variants. We describe methods for preparing such libraries, using serum paraoxonase (PON1). An optimized GFP variant fused to PON1 reported levels of soluble, functional enzyme, enabling selection by flow cytometry and identification of enzyme variants exhibiting improved specific and total activities toward several substrates, including toxic organophosphates.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Aryldialkylphosphatase / analysis*
  • Aryldialkylphosphatase / genetics
  • Aryldialkylphosphatase / metabolism*
  • Directed Molecular Evolution*
  • Genes, Reporter / genetics
  • Genetic Drift
  • Mutation / genetics
  • Peptide Library*

Substances

  • Peptide Library
  • Aryldialkylphosphatase