Related Experiment Video
Updated: Apr 27, 2026
16:02
Quantitative FRET Förster Resonance Energy Transfer Analysis for SENP1 Protease Kinetics Determination
Published on: February 21, 2013
18.0K
Introducing enzyme selectivity: a quantitative parameter to describe enzymatic protein hydrolysis
Claire I Butré1, Stefano Sforza, Harry Gruppen
1Laboratory of Food Chemistry, Wageningen University, Postbus 17, 6700 AA, Wageningen, The Netherlands.
Analytical and Bioanalytical Chemistry
|July 12, 2014
Summary
Enzyme selectivity quantifies how enzymes cleave specific sites in proteins. Researchers measured this for Bacillus licheniformis protease on whey protein, revealing significant differences in hydrolysis rates at various sites.
Area of Science:
- Biochemistry
- Enzymology
- Proteomics
Background:
- Enzymatic protein hydrolysis is crucial in food science and biotechnology.
- Understanding enzyme specificity is key to controlling hydrolysis outcomes.
- Whey protein isolate is a valuable protein source often modified by enzymatic treatment.
Purpose of the Study:
- To introduce and quantify enzyme selectivity as a parameter for describing hydrolysis rates at individual cleavage sites.
- To investigate the specificity of Bacillus licheniformis protease towards beta-lactoglobulin.
- To establish new metrics for assessing peptide identification and quantification quality.
Main Methods:
- Hydrolysis of whey protein isolate using Bacillus licheniformis protease.
- Determination of molar concentration of 58 peptides from beta-lactoglobulin using UV214 signal.
- Calculation of enzyme selectivity based on hydrolysis rates of individual cleavage sites.
- Definition and application of peptide sequence coverage and molar sequence coverage.
Main Results:
- Bacillus licheniformis protease demonstrated high specificity for Glutamic acid (Glu) and Aspartic acid (Asp) residues.
- Enzyme selectivity varied up to a factor of 5,000 among different cleavage sites.
- Average peptide sequence coverage was 94%, and average molar sequence coverage was 75%.
Conclusions:
- Enzyme selectivity provides a quantitative measure of enzyme preference for specific cleavage sites.
- Quantitative discrimination of enzyme preference is essential for understanding enzymatic protein hydrolysis.
- The developed parameters enhance the assessment of peptide identification and quantification in proteomic studies.
Related Concept Videos
01:19Enzyme Kinetics
80.7K
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...
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...
80.7K
01:19Introduction to Enzyme Kinetics
24.3K
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...
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...
24.3K
01:30Enzyme Inhibition
72.3K
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.
72.3K
02:34Enzymes
68.8K
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...
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...
68.8K
01:07Catalytically 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...
Most enzymes...
4.3K
01:13Introduction to Mechanisms of Enzyme Catalysis
9.0K
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...
9.0K