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

Cytoskeletal Proteins in Bacteria01:29

Cytoskeletal Proteins in Bacteria

3.3K
Bacterial cells were initially considered simple, randomly organized structures lacking a cytoskeleton. However, the discovery of cytoskeleton homologs in bacteria led to the change of this opinion. Bacterial cytoskeletal filaments regulate the cell shape, cell polarity, cell division, and partitioning of plasmids during cell division. It was later discovered that bacterial cytoskeletal proteins, mainly actin and tubulin homologs, are diverse compared to their eukaryotic counterparts. On the...
3.3K
Allosteric Proteins-ATCase01:19

Allosteric Proteins-ATCase

5.7K
Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to  N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
5.7K
Microtubule Instability02:17

Microtubule Instability

5.0K
Microtubules are hollow cylindrical filaments having a diameter of approximately 25 nm and a length that varies from 200 nm to 25 μm. GTP-bound tubulin subunits form αβ-heterodimers for microtubule assembly. These core building blocks interact longitudinally, polymerizing into protofilaments. The protofilaments then interact with one another through lateral bonding forces to form stable cylindrical microtubules. These cylindrical filaments are dynamic as they undergo repeated...
5.0K
Protein Complex Assembly02:41

Protein Complex Assembly

10.6K
Proteins can form homomeric complexes with another unit of the same protein or heteromeric complexes with different types.  Most protein complexes self-assemble spontaneously via ordered pathways, while some proteins need assembly factors that guide their proper assembly. Despite the crowded intracellular environment, proteins usually interact with their correct partners and form functional complexes.
Many viruses self-assemble into a fully functional unit using the infected host cell to...
10.6K
Introduction to Actin01:26

Introduction to Actin

5.0K
Actin is a highly conserved cytoskeletal protein found abundantly in eukaryotic cells. It constitutes 10% weight of the total cellular protein in muscle cells, while in non-muscle cells, it is lower and makes up around 1–5 percent of the total cell protein. Actin found in the unicellular amoebae and complex multicellular animals is around 80% similar, demonstrating their conservation over a billion years of evolution.  Actin coding genes are conserved within species and across...
5.0K
Ligand Binding and Linkage00:49

Ligand Binding and Linkage

4.8K
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...
4.8K

You might also read

Related Articles

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

Sort by
Same author

Computer Simulation Modeling for Recreation Management: A Study on Carriage Road Use in Acadia National Park, Maine, USA.

Environmental management·1999
Same author

An Analysis of Higher Order Vibration-Torsion-Rotation Interactions and Spectra for a Molecule with an Internal Rotor.

Journal of molecular spectroscopy·1999
Same author

Preparation of Palladium Ultrafine Particles in Reverse Micelles.

Journal of colloid and interface science·1999
Same author

Prediction of photosynthetic production rate of ethylene using a recombinant cyanobacterium

Journal of theoretical biology·1999
Same author

Conformational Structure of Triblock Copolymers by FT-Raman and FTIR Spectroscopy.

Journal of colloid and interface science·1999
Same author

Surface Characters and Adsorption Behavior of Pb(II) onto a Mesoporous Titanosilicate Molecular Sieve.

Journal of colloid and interface science·1999

Related Experiment Video

Updated: Jun 14, 2025

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments
20:28

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

Published on: October 2, 2012

14.1K

Activity and stability improvement: structure-function insights into CotA from Bacillus subtilis.

Wei He1, Yiru1, Wang1

  • 1Beijing Key Laboratory of Biomass Waste Resource Utilization, College of Biochemical Engieering, Beijing Union University, Beijing 100023, China; Beijing Key Laboratory of Bioactive Substances and Functional Foods, College of Biochemical Engineering, Beijing Union University, Beijing 100023, China.

International Journal of Biological Macromolecules
|June 12, 2025
PubMed
Summary

Researchers engineered a bacterial multicopper oxidase (CotA) variant with 6-fold higher activity and improved thermal stability. Molecular dynamics simulations revealed structural changes linked to enhanced enzyme function and stability for industrial applications.

Keywords:
Catalytic efficiencyCotAMD simulationSite-directed mutagenesisThermal stability

More Related Videos

Monitoring Intraspecies Competition in a Bacterial Cell Population by Cocultivation of Fluorescently Labelled Strains
06:45

Monitoring Intraspecies Competition in a Bacterial Cell Population by Cocultivation of Fluorescently Labelled Strains

Published on: January 18, 2014

8.5K
Using Coculture to Detect Chemically Mediated Interspecies Interactions
08:29

Using Coculture to Detect Chemically Mediated Interspecies Interactions

Published on: October 31, 2013

13.6K

Related Experiment Videos

Last Updated: Jun 14, 2025

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments
20:28

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

Published on: October 2, 2012

14.1K
Monitoring Intraspecies Competition in a Bacterial Cell Population by Cocultivation of Fluorescently Labelled Strains
06:45

Monitoring Intraspecies Competition in a Bacterial Cell Population by Cocultivation of Fluorescently Labelled Strains

Published on: January 18, 2014

8.5K
Using Coculture to Detect Chemically Mediated Interspecies Interactions
08:29

Using Coculture to Detect Chemically Mediated Interspecies Interactions

Published on: October 31, 2013

13.6K

Area of Science:

  • Biochemistry
  • Structural Biology
  • Enzyme Engineering

Background:

  • Bacterial multicopper oxidase (CotA) is a versatile enzyme with potential in industrial biocatalysis and environmental remediation.
  • Existing methods to enhance CotA's properties include directed evolution and protein design, but structural insights remain limited.

Purpose of the Study:

  • To investigate the structural basis for enhanced activity and stability in CotA mutants.
  • To identify specific mutations that improve CotA's performance for industrial applications.

Main Methods:

  • Site-directed mutagenesis was used to create CotA single and double mutants from Bacillus subtilis.
  • Molecular dynamics (MD) simulations were employed to analyze the structural features of the mutants.
  • Enzyme activity and thermal stability were experimentally assessed.

Main Results:

  • The T377I/T418G CotA variant showed a 6.12-fold increase in ABTS-specific activity and 50% improved thermal stability at 80°C.
  • MD simulations indicated that increased activity correlated with an expanded enzyme binding pocket.
  • Enhanced thermal stability was associated with a higher proportion of random coils in the enzyme's secondary structure.

Conclusions:

  • Specific mutations can significantly enhance CotA's catalytic activity and thermal stability.
  • Structural analysis via MD simulations provides insights into the mechanisms underlying enzyme improvement.
  • This study provides a foundation for optimizing CotA for diverse industrial and environmental applications.