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Related Concept Videos

Enzymes02:34

Enzymes

69.1K
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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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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Enzyme Kinetics01:19

Enzyme Kinetics

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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...
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Introduction to Enzyme Kinetics01:19

Introduction to Enzyme Kinetics

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Enzyme kinetics studies the rates of biochemical reactions. Scientists monitor the reaction rates for a particular enzymatic reaction at various substrate concentrations. Additional trials with inhibitors or other molecules that affect the reaction rate may also be performed.
The experimenter can then plot the initial reaction rate or velocity (Vo) of a given trial against the substrate concentration ([S]) to obtain a graph of the reaction properties. For many enzymatic reactions involving a...
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Enzymes and Activation Energy01:13

Enzymes and Activation Energy

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Enzymes and Activation Energy01:13

Enzymes and Activation Energy

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The activation energy (or free energy of activation), abbreviated as Ea, is the small amount of energy input necessary for all chemical reactions to occur. During chemical reactions, certain chemical bonds break, and new ones form. For example, when a glucose molecule breaks down, bonds between the molecule's carbon atoms break. Since these are energy-storing bonds, they release energy when broken. However, the molecule must be somewhat contorted to get into a state that allows the bonds to...
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Related Experiment Video

Updated: May 6, 2026

Determination of Microbial Extracellular Enzyme Activity in Waters, Soils, and Sediments using High Throughput Microplate Assays
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Determination of Microbial Extracellular Enzyme Activity in Waters, Soils, and Sediments using High Throughput Microplate Assays

Published on: October 1, 2013

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Analysis of enzyme activities.

Guillaume Ménard1, Benoit Biais, Duyên Prodhomme

  • 1UMR 1332, INRA, Villenave d'Ornon cedex, France.

Methods in Molecular Biology (Clifton, N.J.)
|November 14, 2013
PubMed
Summary
This summary is machine-generated.

Microplate technology allows for the efficient measurement of over 50 enzyme activities, crucial for modeling biochemical pathways. This study presents protocols for UDP-glucose pyrophosphorylase and pyrophosphate:fructose-6-phosphate 1-phosphotransferase assays.

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Area of Science:

  • Biochemistry
  • Metabolic Engineering
  • Enzymology

Background:

  • Enzyme activity evaluation is vital for biochemical pathway modeling.
  • Microplate technology offers high-throughput analysis of numerous enzymes in diverse biological samples.
  • Central metabolic pathways can be comprehensively studied using this approach.

Purpose of the Study:

  • To highlight the utility of microplate technology for enzyme activity determination.
  • To present detailed protocols for specific enzyme assays.
  • To demonstrate the application of these methods in biological research.

Main Methods:

  • Utilizing microplate technology for enzyme activity quantification.
  • Employing direct, indirect, and highly sensitive assay formats.
  • Implementing real-time assays for enzyme kinetics.

Main Results:

  • Demonstrated the capacity to measure >50 enzyme activities per microplate.
  • Provided validated protocols for UDP-glucose pyrophosphorylase (E.C. 2.7.7.9) and pyrophosphate:fructose-6-phosphate 1-phosphotransferase (E.C. 2.7.1.90).
  • Showcased the applicability across various biological materials.

Conclusions:

  • Microplate technology is a powerful tool for comprehensive enzyme analysis.
  • The presented protocols facilitate the study of key enzymes in central metabolism.
  • This methodology supports advancements in metabolic pathway modeling and biological research.