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 Concept Videos

Enzyme Kinetics01:19

Enzyme Kinetics

81.0K
Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
81.0K
Catalytically Perfect Enzymes01:07

Catalytically Perfect Enzymes

4.3K
The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at high-speed. Their catalytic efficiency values range from 108-109 M-1s-1. These enzymes are also called 'diffusion-controlled' as the only rate-limiting step in the catalysis is that of the substrate diffusion into the active site. Examples include triose phosphate isomerase, fumarase, and superoxide dismutase.
 
Most enzymes...
4.3K
Enzyme Inhibition01:30

Enzyme Inhibition

72.4K
Inhibitors are molecules that reduce enzyme activity by binding to the enzyme. In a normally functioning cell, enzymes are regulated by a variety of inhibitors. Drugs and other toxins can also inhibit enzymes. Some inhibitors bind to the enzyme’s active site, while others inhibit enzymatic activity by binding to other sites on the protein structure.
72.4K
Enzymes02:34

Enzymes

68.9K
Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...
68.9K

You might also read

Related Articles

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

Sort by
Same author

CASP11 refinement experiments with ROSETTA.

Proteins·2015
Same author

Structure of a designed tetrahedral protein assembly variant engineered to have improved soluble expression.

Protein science : a publication of the Protein Society·2015
Same author

Unique double-ring structure of the peroxisomal Pex1/Pex6 ATPase complex revealed by cryo-electron microscopy.

Proceedings of the National Academy of Sciences of the United States of America·2015
Same author

Mechanistic Analysis of an Engineered Enzyme that Catalyzes the Formose Reaction.

Chembiochem : a European journal of chemical biology·2015
Same author

Design of ordered two-dimensional arrays mediated by noncovalent protein-protein interfaces.

Science (New York, N.Y.)·2015
Same author

Designing Two-Dimensional Protein Arrays through Fusion of Multimers and Interface Mutations.

Nano letters·2015

Related Experiment Video

Updated: May 4, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

7.7K

Computationally designed libraries for rapid enzyme stabilization.

Hein J Wijma1, Robert J Floor, Peter A Jekel

  • 1Department of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.

Protein Engineering, Design & Selection : PEDS
|January 10, 2014
PubMed
Summary
This summary is machine-generated.

Computational design created a library of enzyme variants with significantly improved stability and function. This Framework for Rapid Enzyme Stabilization by Computational libraries (FRESCO) method enhances protein engineering efficiency.

Keywords:
enzyme stabilityin silico designin silico screeningprotein stability engineeringthermostability

More Related Videos

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems
07:35

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems

Published on: June 14, 2021

2.3K
Development of Inhibitors of Protein-protein Interactions through REPLACE: Application to the Design and Development Non-ATP Competitive CDK Inhibitors
10:33

Development of Inhibitors of Protein-protein Interactions through REPLACE: Application to the Design and Development Non-ATP Competitive CDK Inhibitors

Published on: October 26, 2015

10.9K

Related Experiment Videos

Last Updated: May 4, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

7.7K
Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems
07:35

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems

Published on: June 14, 2021

2.3K
Development of Inhibitors of Protein-protein Interactions through REPLACE: Application to the Design and Development Non-ATP Competitive CDK Inhibitors
10:33

Development of Inhibitors of Protein-protein Interactions through REPLACE: Application to the Design and Development Non-ATP Competitive CDK Inhibitors

Published on: October 26, 2015

10.9K

Area of Science:

  • Protein Engineering
  • Computational Biology
  • Biochemistry

Background:

  • Enzyme engineering for enhanced stability has broad applications.
  • Mesostable enzymes like limonene epoxide hydrolase are targets for improvement.

Purpose of the Study:

  • To computationally design and experimentally validate stabilized variants of limonene epoxide hydrolase.
  • To develop a rapid computational framework for enzyme stabilization.

Main Methods:

  • In silico selection of point mutations and disulfide bonds based on predicted folding free energy and flexibility.
  • Orthogonal screening to remove chemically unfavorable mutations.
  • Experimental validation of computationally designed variants.

Main Results:

  • Identified 21 stabilizing mutations in the enzyme.
  • Developed multi-site mutants with melting temperatures increased from 50°C to 85°C.
  • Achieved enhanced catalytic activity, preserved regioselectivity, and a >250-fold longer half-life.

Conclusions:

  • The Framework for Rapid Enzyme Stabilization by Computational libraries (FRESCO) enables efficient engineering of highly stable and functional enzymes.
  • This computational approach significantly reduces screening efforts compared to traditional directed evolution.