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Related Experiment Videos

In vitro selection for enzymatic activity: a model study using adenylate cyclase.

Heike Strobel1, Daniel Ladant, Jean-Luc Jestin

  • 1Unité de Chimie Organique, Institut Pasteur, 28 rue du Dr Roux, 75724 Paris 15, France.

Journal of Molecular Biology
|August 30, 2003
PubMed
Summary

A novel in vitro enzyme selection method using phage display and antibody fragments was developed. This technique enables the selection of enzymes for various chemical reactions, exemplified by adenylate cyclase activity.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzyme Engineering

Background:

  • Enzyme selection is crucial for biocatalysis and synthetic biology.
  • Developing versatile in vitro selection methods is essential for discovering novel enzymatic functions.
  • Antibody-based selection offers a powerful approach for targeting specific reaction products.

Purpose of the Study:

  • To describe a generalizable in vitro enzyme selection method.
  • To demonstrate the utility of filamentous phage display in enzyme discovery.
  • To validate the approach using Bordetella pertussis adenylate cyclase.

Main Methods:

  • Utilized filamentous phage display technology.
  • Engineered a tailor-made antibody fragment targeting the reaction product.

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  • Applied the method to select for enzymes catalyzing ATP to cyclic AMP conversion.

    • Main Results:

    • Successfully demonstrated an in vitro enzyme selection system.
    • The method is adaptable for a wide range of chemical reactions.
    • The specific example showed successful selection related to adenylate cyclase activity.

    Conclusions:

    • The described phage display and antibody-based selection is a versatile tool for enzyme discovery.
    • This method can be broadly applied to various enzymatic transformations.
    • It offers a promising avenue for engineering enzymes with desired catalytic activities.