Module 1: Principles of enzymology
1) Historical background. Nomenclature and classification of enzymes. Determination of enzyme activity. Enzyme function, active sites, cofactors, pecificity
2) Reversible intramolecular forces
3) The formation of enzyme-substrate complex and molecular recognition
4) Structural fluctuation and molecular dynamics in enzyme catalysis
5) The basic principles and key mechanisms of enzymatic catalysis
6) Thermodynamics and structure-catalysis relationships
Module2: Enzyme kinetics
1) The principles of enzyme kinetics and the factors affecting the catalytic activity.
2) Kinetic parameters and reaction equilibrium
3) The analysis of kinetic data, Michaelis-Menten equation and methods of plotting enzyme kinetics data
4) Effect of pH and temperature on enzyme stability and activity.
5) The principles of enzyme inhibition, types of inhibition and the concepts of allosteric activators or inhibitors. Reversible and irreversible inhibition (inactivation). Inhibition constants. Interaction of enzymes and xenobiotic compounds (drugs, insecticides, herbicides, etc.)
6) Multi-substrate enzyme reactions
7) Isotopes in enzyme reaction rate determination
8) Mechanobiology of enzyme systems
Module 3. Enzyme engineering
1) The principles of designing structural modifications using biocomputing methods and recombinant DNA technology
2) Molecular methods for site-directed mutagenesis and random mutagenesis.
3) Principles and methods of in vitro directed molecular evolution
4) High-throughput screening methods for enzyme selection
5) De novo design of new functional enzymes
6) Chemical modification of enzyme structure
7) Paleoenzymology and reconstruction of ancient enzymes.
8) Hybrid enzymes, semisynthetic enzymes, artificial enzymes, catalytic antibodies and ribozymes
9) Enzyme nanomachines and multi-complex enzymes
10) Applications of engineered enzymes in agriculture, medicine, industry and environmental technologies. Enzymes for molecular biology (structure, mechanism, applications)
Module 4: Enzyme applications
1) Εnzymes that recognize and modify nucleic acids
2) Enzymes as molecular targets for drug design
3) Detoxifying enzymes (oxygenases, transferases, hydrolases, etc.)
Learning outcomes
Bibliography
1) Ιωάννης Κλώνης (2007) Ενζυμολογία, Έμβρυο.
2) Yon-Kahn, Jeannine, Hervé, G. (2010) Molecular and Cellular Enzymology. Springer USA.
3) Hans Bisswanger (2011) Practical Enzymology, 2nd Edition, Wiley-Blackwell.
4) Sheldon J. Park, Jennifer R. Cochran (2010) Protein Engineering and Design. Taylor and Francis
Group.
5) Stefan Lutz, Uwe T. Bornscheuer (2011) Protein Engineering Handbook, Volume 1 & Volume 2,
Wiley-VCH Verlag GmbH & Co. KGaA.
Biotechnology is a rapidly advancing discipline which aims at exploitting the progress in life and physical sciences as well as other related fields, in developing new and advanced products, processes and services
