Ultra-high-throughput screening based on cell-surface display and fluorescence-activated cell sorting for the identification of novel biocatalysts.

Stefan Becker; Hans-Ulrich Schmoldt; Thorsten Michael Adams; Susanne Wilhelm; Harald Kolmar

doi:10.1016/j.copbio.2004.06.001

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Ultra-high-throughput screening based on cell-surface display and fluorescence-activated cell sorting for the identification of novel biocatalysts.

Stefan Becker·Hans-Ulrich Schmoldt·Thorsten Michael Adams·Susanne Wilhelm·Harald Kolmar

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Abstract

Enzyme libraries displayed on the surface of microbial cells or microbeads can be screened with fluorogenic substrates that provide a physical linkage of the reaction product to the corresponding enzyme. Libraries exceeding 10(9) different variants can be quantitatively analysed and screened by flow cytometry at a rate of 30 000 cells/second. The promise of screening methods based on fluorescence-activated cell sorting for directed enzyme evolution is being realized and significantly improved enzymes have been reported recently.

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Bacteria Biological Assay Catalysis Enzyme Activation Enzymes Flow Cytometry Fluorescence Resonance Energy Transfer Membrane Proteins Peptide Library Protein Engineering Protein Interaction Mapping Journal Article Review

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Stefan Becker, Hans-Ulrich Schmoldt, Thorsten Michael Adams, Susanne Wilhelm, Harald Kolmar. 2004. Ultra-high-throughput screening based on cell-surface display and fluorescence-activated cell sorting for the identification of novel biocatalysts.. https://doi.org/10.1016/j.copbio.2004.06.001

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