Directed enzyme evolution : screening and selection methods /

Bibliographic Details
Other Authors: Arnold, Frances Hamilton, Georgiou, George
Format: Book
Language:English
Published: Totowa, N.J. : Humana Press, [2003]
Series:Methods in molecular biology (Clifton, N.J.) ; v. 230.
Subjects:
Table of Contents:
  • Genetic complementation protocols
  • Use of pol I-deficient E. coli for functional complementation of DNA polymerase
  • Selection of novel eukaryotic DNA polymerases by mutagenesis and genetic complementation of yeast
  • Autogene selections
  • Selection for soluble proteins via fusion with chloramphenicol acetyltransferase
  • Proside : a phage-based method for selecting thermostable proteins
  • Minimization of proteins by random fragmentation and selection
  • Evaluating a screen and analysis of mutant libraries
  • Screening mutant libraries in Saccharomyces cerevisiae
  • Solid-phase screening using digital image analysis
  • Screening for thermostability
  • High-throughput screening of mutant α-amylase libraries for increased activity at 120⁰C
  • High-throughput carbon monoxide binding assay for cytochromes P450
  • High-throughput screen for aromatic hydroxylation
  • Colorimetric screen for aliphatic hydroxylation by cytochrome P450 using p-nitrophenyl-substituted alkanes
  • High-throughput screens based on NAD(P)H depletion
  • High-throughput tetramethylbenzidine (TMB) screen for peroxidases
  • Screen for oxidases by detection of hydrogen peroxide with horseradish peroxidase
  • Colorimetric dehydrogenase screen board on NAD(P)H generation
  • Colorimetric assays for screening laccases
  • pH sensing agar plate assays for esterolytic enzyme activity
  • A pH-indicator-based screen for hydrolytic haloalkane dehalogenase
  • Detection of aromatic α-hydroxyketones with tetrazolium salts
  • Selection of heat-stable Clostridium cellulovorans cellulases
  • Screening and selection strategies for disulfide isomerase activity
  • An overview of high-throughput screening systems for enantioselective enzymatic transformations
  • Select protocols of high-throughput ee-screening systems for enantioselective enzymatic transformations
  • Directed evolution of the substrate specificities of a site-specific recombinase and an aminoacyl-tRNA synthetase using fluorescence-activated cell sorting (FACS)
  • Calmodulin-tagged phage and two-filter sandwich assays for the identification of enzymatic activities
  • High-throughput FACS method for directed evolution of substrate specificity
  • Improving protein folding efficiency by directed evolution using the GFP folding reporter