After completing this course the students should have general knowledge on the molecular nature and function of enzymes. The students should understand the role of enzymes in the cell and be able to describe the different mechanisms involved in the regulation of their activity.
The students should be able to construct mathematical models to describe the kinetic behavior of enzymes under different situations. The models might include the presence of inhibitors, one or two substrates, pH dependence, and cooperativity. The students should know how to use the models to analyze experimental data and determine the kinetic parameters that are relevant in each case, understanding their physical significance. Also, they should be aware of the assumptions used in the derivation of the models and the experimental conditions that guarantee their validity.
Maria Teresa Nunes Mangas Catarino
Weekly - 4
Total - 56
There are no requirements, but this curricular unit relies on previous knowledge of general concepts of Biochemistry.
- Fundamentals of Enzymology. 1999 (3rd Edition) Nicholas C. Price and Lewis Stevens,OxfordUniversity Press
- Cinética Enzimática. 2006. Teresa Moura e Francisco Pinto (Edição do Departamento de Química da FCT/UNL)
- Biochemistry. 2006 (6th edition) Jeremy M. Berg, John L. Tymoczko, L. Stryer, Ed. W.H. Freeman & Company
- Fundamentals of Enzyme Kinetics. 2004 (3rd Edition) Athel Cornish-Bowden,Portland Press
- Enzyme Kinetics : Behavior and Analysis of Rapid Equilibrium and Steady-State Enzyme Systems. 1993. Irwin H. Segel, Wiley Classics Library Edition
The teaching is organized in lectures, problem solving workshops and discussion of experimental results. The contents of the course will be presented to the students during the lectures. The application of the concepts and mathematical models will be made in the workshops where the students will work in small groups. In some classes the groups will be called to present and discuss a particular problem of enzyme kinetics.
Final Grade = 70% (2 written examination papers) + 20% oral examination + 10% presentation
The student has to get a final grade of at least 9.5/20 to pass the course. The average grade of the written papers cannot be lower than 9.5/20.
To be evaluated the student has to attend at least 14 classes (lectures + workshops) and has to be present at the oral examination.
1. General concepts: properties of enzymes and their role in the cell. Cofactors and coenzymes.
2. The active site. Transition state theory. Effect of temperature and pH.
3. Molecular mechanisms of enzyme action.
4. Regulation of enzymatic activity.
5. Application of enzymes in industry and health
6. Enzyme kinetics.
6.1. Derivation of the Michaelis-Menten equation and its assumptions. Determination of the kinetic parameters of the enzyme: Vmax and KM.
6.2. Enzyme kinetics in the presence of a reversible inhibitor: model of Webb.
6.3. Enzyme kinetics for reactions involving two substrates.
6.4. pH dependence of enzymatic activity. Kinetic models with one or more acid/base centres. Determination of pKa values.
6.5. Oligomeric enzymes with and without cooperativity. Kinetic models for cooperativity: Monot Wyman and Changeux (MWC) and Koshland (induced fit). Measurement of cooperativity.