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Related Concept Videos

Introduction to Mechanisms of Enzyme Catalysis01:13

Introduction to Mechanisms of Enzyme Catalysis

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For many years, scientists thought that enzyme-substrate binding took place in a simple "lock-and-key" fashion. This model stated that the enzyme and substrate fit together perfectly in one instantaneous step. However, current research supports a more refined view scientists call induced fit. The induced-fit model expands upon the lock-and-key model by describing a more dynamic interaction between enzyme and substrate. As the enzyme and substrate come together, their interaction causes...
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Enzymes02:34

Enzymes

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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...
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Induced-fit Model01:13

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Most chemical reactions in cells require enzymes—biological catalysts that speed up the reaction without being consumed or permanently changed. They reduce the activation energy needed to convert the reactants into products. Enzymes are proteins, that usually work by binding to a substrate—a reactant molecule that they act upon.
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Enzyme Inhibition01:30

Enzyme Inhibition

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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.
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Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

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Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
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Ligand Binding and Linkage00:49

Ligand Binding and Linkage

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Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked.  In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...
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Related Experiment Video

Updated: Apr 20, 2026

Hydrophobic Salt-modified Nafion for Enzyme Immobilization and Stabilization
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Hydrophobic Salt-modified Nafion for Enzyme Immobilization and Stabilization

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Enhanced enzyme stability through site-directed covalent immobilization.

Jeffrey Chun Yu Wu1, Christopher Hayden Hutchings1, Mark Jeffrey Lindsay1

  • 1Department of Chemical Engineering, Brigham Young University, Provo, UT, USA.

Journal of Biotechnology
|December 3, 2014
PubMed
Summary

Controlled enzyme immobilization using the PRECISE system enhances enzyme activity and stability. This method offers significant improvements over traditional techniques, reducing costs and boosting biocatalysis.

Keywords:
BiocatalysisCell-free protein synthesis.Enzyme immobilizationEnzyme stabilityGreen manufacturingNon-canonical amino acids

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Area of Science:

  • Biocatalysis
  • Protein Engineering
  • Biotechnology

Background:

  • Enzyme immobilization advances reduce costs and boost biocatalysis but face challenges like leaching and poor enzyme stability.
  • Current methods lack control over enzyme orientation, limiting performance.
  • Directed immobilization is crucial for optimizing enzyme function.

Purpose of the Study:

  • To investigate the impact of enzyme orientation on activity and stability using a novel immobilization system.
  • To demonstrate the benefits of site-specific covalent immobilization over random methods.

Main Methods:

  • Utilized the PRECISE (Protein Residue-Explicit Covalent Immobilization for Stability Enhancement) system for directed enzyme immobilization.
  • Incorporated non-canonical amino acids and employed click chemistry for covalent attachment.
  • Tested T4 lysozyme immobilized at various locations relative to the active site under denaturing conditions.

Main Results:

  • PRECISE system enabled true covalent immobilization with controlled enzyme orientation.
  • Immobilized enzymes exhibited enhanced activity and stability compared to randomly immobilized counterparts.
  • PRECISE immobilized enzymes showed 50% and 73% greater activity after freeze-thaw and chemical denaturant treatments, respectively.

Conclusions:

  • Orientation control of covalently immobilized enzymes offers significant advantages in activity and stability.
  • The PRECISE system provides a powerful tool for enhancing enzyme performance in biocatalysis.
  • This method overcomes limitations of traditional immobilization techniques, paving the way for more efficient enzyme applications.