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Novel methods for biocatalyst screening.

D Wahler1, J L Reymond

  • 1Departement für Chemie und Biochemie, Universität Bern, Switzerland.

Current Opinion in Chemical Biology
|April 3, 2001
PubMed
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Recent advances in catalysis assays enable high-throughput screening of biocatalyst libraries. New methods, including fluorogenic substrates, allow for enantioselective and stereoselective enzyme assays.

Area of Science:

  • Biocatalysis and enzyme engineering
  • Analytical chemistry and assay development

Background:

  • High-throughput screening is crucial for identifying novel biocatalysts.
  • Existing assay methods face limitations in speed and scope for library screening.

Purpose of the Study:

  • To review recent advances in catalysis assays for high-throughput screening of biocatalyst libraries.
  • To highlight the capabilities of various instrumental and reagent-based assay formats.

Main Methods:

  • Instrumental assays: High-performance liquid chromatography (HPLC), mass spectrometry (MS), capillary electrophoresis (CE), IR-thermography.
  • Reagent-based assays: Spectroscopic signals (UV/VIS, fluorescence).
  • Substrate-based assays: Fluorogenic and chromogenic substrates.

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Main Results:

  • Developed instrumental assays offer precise reaction monitoring.
  • Spectroscopic and substrate-based assays provide sensitive detection of enzyme activity.
  • Fluorogenic substrates enable enantioselective and stereoselective assays for diverse enzymes.

Conclusions:

  • Recent advancements have significantly improved high-throughput screening of biocatalysts.
  • A variety of assay techniques are available, catering to different enzyme classes and screening needs.
  • Fluorogenic substrates are particularly valuable for stereoselective enzyme characterization.