ENZYMOLOGY
cod. 04241

Academic year 2016/17
1° year of course - Second semester
Professor
Davide FERRARI
Academic discipline
Biochimica (BIO/10)
Field
Discipline del settore biomolecolare
Type of training activity
Characterising
42 hours
of face-to-face activities
6 credits
hub: PARMA
course unit
in - - -

Learning objectives

knowledge of enzymatic reactions and their application in industry
Students must show to have: knowledge and understanding, applying knowledge and understanding, making judgements, communication skills, learning skills

Prerequisites

none

Course unit content

Enzymes: Background. Nomenclature and classification of enzymes. Biological catalysts.
Mechanism of action of enzymes: Relationships protein structure / function enzyme. The enzyme-substrate complex, induced fit. Theory of the transition state. Catalysis. Acid-base catalysis, covalent catalysis, metal ion catalysis, electrostatic catalysis. Effect of proximity and orientation. Stabilization of the transition state. Coenzymes, vitamins and essential metals. Examples of enzyme catalytic mechanisms (ie: Serin protease; Cysteine proteases; Aspartate proteases, etc.). Experimental methods for the study of enzyme mechanisms.

Enzyme kinetics: Chemical Kinetics. Order of the reactions. Michaelis-Menten equation and its derivation at steady state. Calculation of Vmax and Km Meaning of the kinetic constants Km and Kcat. Reaction rate. Graphical methods for the determination of Km and Vmax. Reactions to most substrates. Allosteric enzymes, cooperativity and sigmoidal kinetics. Effect of pH and temperature on enzyme activity. State pre-stationary.

Enzyme inhibition: Reversible inhibitors: competitive inhibitors, non-competitive and incompetitivi. Graphical methods for the identification of the mechanism of inhibition. Examples of enzyme inhibitors for pharmacological use.

Enzyme regulation: regulation of synthesis and degradation of enzymes.

Applied enzymology: Immobilization of enzymes, various types of immobilized enzymes. Most common reactions for the immobilization of enzymes on insoluble matrices or by using bifunctional reagents. Biosensors. Industrial applications of enzymes (notes). Enzymes as analytical tools. ELISA methods.

Full programme

Nomenclature and classification of enzymes. Biological catalysts.
Mechanism of action of enzymes: Relationships protein structure / function enzyme. The enzyme-substrate complex, induced fit. Theory of the transition state. Catalysis. Acid-base catalysis, covalent catalysis, metal ion catalysis, electrostatic catalysis. Effect of proximity and orientation. Stabilization of the transition state. Coenzymes, vitamins and essential metals. Examples of enzyme catalytic mechanisms (ie: Serin protease; Cysteine proteases; Aspartate proteases, etc.). Experimental methods for the study of enzyme mechanisms.

Enzyme kinetics: Chemical Kinetics. Order of the reactions. Michaelis-Menten equation and its derivation at steady state. Calculation of Vmax and Km Meaning of the kinetic constants Km and Kcat. Reaction rate. Graphical methods for the determination of Km and Vmax. Reactions to most substrates. Allosteric enzymes, cooperativity and sigmoidal kinetics. Effect of pH and temperature on enzyme activity. State pre-stationary.

Enzyme inhibition: Reversible inhibitors: competitive inhibitors, non-competitive and incompetitivi. Graphical methods for the identification of the mechanism of inhibition. Examples of enzyme inhibitors for pharmacological use.

Enzyme regulation: regulation of synthesis and degradation of enzymes.

Applied enzymology: Immobilization of enzymes, various types of immobilized enzymes. Most common reactions for the immobilization of enzymes on insoluble matrices or by using bifunctional reagents. Biosensors. Industrial applications of enzymes (notes). Enzymes as analytical tools. ELISA methods.

Bibliography

Enzymes: A Practical Introduction to Structure, Mechanism, and Data Analysis
R. A. Copeland

Teaching methods

frontal teaching

Assessment methods and criteria

written examination
Students must show to have: knowledge and understanding, applying knowledge and understanding, making judgements, communication skills, learning skills

Other information

none

2030 agenda goals for sustainable development

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