Elements of biochemistry and biochemical methods
Learning outcomes of the course unit
In the first part of the course, the notions concern the structure and the multiple functions of proteins, whose crucial roles played in cellular processes will be highlighted, as well as composition, structural organization and transport properties of biological membranes. Students will also be introduced to the knowledge of the major classes of small biological molecules (carbohydrates, lipids, amino acids) and will learn the basic metabolic pathways by which these molecules are absorbed, degraded and used by the body.
In the second part of the course, students will achieve information on the main experimental techniques used in the biochemical laboratory, mainly directed to the identification and isolation of proteins, as well as on the use of antibodies and enzymes for analytical purposes.
In order to address the topics of the course "Elements of Biochemistry and Biochemical Methodologies", students should possess basic knowledge of Chemistry and Organic Chemistry.
Course contents summary
Structure and function of proteins. The amino acids. The peptide bond. The levels of the structural organization of proteins: from the primary to the quaternary structure. Enzymes as fundamental catalysts of biological reactions. Role of enzymes in digestion and assimilation of proteins, carbohydrates and lipids. Hemoglobin and myoglobin: role in the oxygen cycle and structural and functional properties.
Biological membranes. The membrane lipids, structural organization of biological membranes, active and passive transport of solutes (endogenous compounds and xenobiotics) across membranes.
The nutrients, their assimilation and their metabolic transformations, related to the extraction of energy essential for cell life and to biosyntheses: aerobic and anaerobic glycolysis, fermentations, Krebs cycle, beta-oxidation of fatty acids, amino acid catabolism, mitochondrial respiratory chain.
Basic properties of proteins related to Biochemical Methodologies: isoelectric point (pI) of proteins, protein solubility as a function of pH and ionic strength; acidic and thermal protein denaturation and denaturation by detergents; intrinsic absorption and fluorescence properties of proteins.
Basic methodologies dedicated to DNA and proteins for both analytical and preparative purposes: electrophoretic and chromatographic techniques. Antibodies: structure-function relationship and immune response; preparation of monoclonal and polyclonal antibodies; use of antibodies as analytical reagents in immunochemical techniques (ELISA methods and Western Blotting). Outline of methods of determination of enzyme activity; reversible and irreversible enzyme inhibitors, and their biotechnological relevance (drugs, pesticides ...).
Biomolecules and Metabolism
Nelson , Cox. Introduzione alla Biochimica di Lehninger, Zanichelli Editore
Bonaccorsi, Contestabile, Di Salvo. Metodologie Biochimiche, Casa Editrice Ambrosiana.
Oral lectures, with the use of multimedia means. There will also be visits to the biochemical laboratory, during which students will have the opportunity to view the use of basic techniques regarding: extraction of proteins and DNA from the cells; chromatographic separation of proteins; electrophoretic analysis of proteins and DNA; spectrophotometric analysis of protein.
Assessment methods and criteria
The acquired notions and the ability to use them in practice will be verified through a written examination regarding the structure and function of proteins, the properties of biological membranes and the metabolism, and through a final oral examination regarding the biochemical methodologies. In these tests the ability to present clearly ideas and analysis will be evaluated.