Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Simultaneous optimization of enzyme activity and quaternary structure by directed evolution.

Katherina Vamvaca1, Maren Butz, Kai U Walter

  • 1Laboratorium für Organische Chemie, Swiss Federal Institute of Technology, ETH Hönggerberg, CH-8093 Zurich, Switzerland.

Protein Science : a Publication of the Protein Society
|July 1, 2005
PubMed
Summary

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

From First Principles to Function: How AI Is Reshaping Enzyme Design.

Biochemistry·2026
Same author

An engineered closed-shell, two-component, 480-subunit nucleocapsid.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

FEMA GRAS assessment of natural flavor complexes: Vanilla extract, Bitter almond oil, Wintergreen oil and related flavoring ingredients.

Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association·2026
Same author

FEMA GRAS assessment of natural flavor complexes: Pepper, ginger, coniferous-derived and related flavoring ingredients.

Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association·2025
Same author

Identification of overoxidizing and non-overoxidizing NAD-dependent methanol dehydrogenases and implications for synthetic methylotrophy.

Nature communications·2025
Same author

Computational design of cysteine proteases.

bioRxiv : the preprint server for biology·2025

Directed evolution enhanced enzyme efficiency 400-fold. Laboratory-evolved chorismate mutase gained novel quaternary states and optimized activity through targeted mutations, showcasing protein engineering potential.

Area of Science:

  • Biochemistry
  • Protein Engineering
  • Directed Evolution

Background:

  • Natural enzymes exhibit diverse structures and high efficiency.
  • Engineered chorismate mutase possessed low activity and an unusual hexameric structure.

Purpose of the Study:

  • To augment the efficiency of an engineered chorismate mutase.
  • To investigate the effects of directed evolution on enzyme structure and function.

Main Methods:

  • Utilized two rounds of DNA shuffling and genetic selection.
  • Performed detailed biophysical characterization of the evolved enzyme variant.

Main Results:

  • Achieved a 400-fold increase in enzyme efficiency.
  • Identified three non-active-site mutations responsible for the enhancement.

Related Experiment Videos

  • Observed the evolved variant predominantly exists as a trimer in solution, with similar stability to the parent hexamer.

    • Conclusions:

    • Directed evolution is a powerful tool for optimizing enzyme activity.
    • Small, non-active-site mutations can drastically alter protein structure and function.
    • Protein-protein interactions can be modified to create novel quaternary states with improved catalytic properties.