INTRODUCTION TO ENZYMATIC ANALYSIS – Session in English
Biotechnologies
Objectifs
Understand the basic principles of enzymatic analysis and its applications.
Become familiar with common methods and techniques used to measure enzyme activity.
Interpret and critically assess enzymatic assay results in a laboratory context.
Contenu pédagogique
THEORY
- Enzymes
- Structure and the active site
- The enzyme–substrate complex
- Enzyme–substrate reaction
- Concept of initial velocity
- The Michaelis–Menten model
- Kinetic parameters of a single-substrate reaction: graphical determination
- Effectors of enzymatic reactions
- Influence of pH and temperature
- Enzyme inhibition: basic concepts
- Kinetic parameters of inhibition
- Methods for measuring enzymatic activity
- Substrate assays
- General principles: endpoint assays, kinetic assays
- Review of selected protocols (case studies)
PRACTICAL WORK
- Kinetics
- Substrate assays
- Measurement of enzyme activity