BIOCHEMISTRY WITH LAB.
Acquisition of knowledge of the universal molecular components of cells, with particular regard to their biological function. Moreover, the students will be able to describe the biochemical pathways involved on the energy transduction. Finally, the student will get the knowledge of the basic techniques for isolation and purification of proteins.
The student will be able to disseminate, to not expert audience, what are the main biomolecules, their biological role and their involvement in the primary metabolism.
The student will be able to access the main literature databases in the field of biochemistry, in order to update the acquired knowledge and to get more insights in the most recent issues in basic biochemistry.
The knowledge of the following topics, derived by the “General and Inorganic Chemistry” and “Organic Chemistry” courses are required:
Periodical properties of the elements (electronegativity, metal and non metals, et cetera)
Definitions of pH and buffer solutions
Basic knowledge of reaction kinetics (activation energy, reaction coordinate diagrams)
Colligative properties and osmosis
Properties and reactions of main organic molecules (alkanes, carboxylic acids and their derivatives, amines, aromatic compounds)
Types of isomers: conformation, configuration, chiral compounds.
Basic knowledge of stereochemistry.
The cellular theory and the molecular bases of life. The structure of eukaryotic and prokaryotic cells.
The proteins: structure and properties of standard aminoacids. The primary, secondary, tertiary and quaternary structures. Interaction with ligands and allosteric proteins. The enzymes, the Michaelis - Menten kinetics, the enzyme inhibition, the allosteric enzymes.
Experimental methods for isolation and purification of proteins. The carbohydrates: mono-, oligo- and polysaccharides.
The simple and compound lipids; cellular membranes. The metabolism: thermodynamics and bioenergetics. Hydrolysis of the phosphorylated compounds. The redox processes in the biological systems and the electrons' carrier molecules.
The chemical reactions, the enzymes regulation and the energy balance of: Glycolysis and Gluconeogenesis; alcoholic and lactic fermentation; pentose phosphate pathway; glycogen metabolism; Krebs cycle; beta-oxidation of fatty acids; Biosynthesis of fatty acids;
The electron transport chain and the oxidative phosphorylation: electron carrier molecules and complexes of the respiratory chain. The chemiosmotic model and the synthesis of ATP.
The protein synthesis and the genetic code.
Block 1: isolation and purification of proteins. (12 hours classroom lesson) Topics: Chromatography techniques. Purity criteria of proteins. Centrifugation. Isoelectrofocusing. Immunochemistry. Proteins’ concentration determination.
Block 2: Enzymes’ activity assays (6 hours classroom lesson) Topics: Biological assays for enzymes’ activity. Fluorescence and luminometry applications.
Block 3: Proteins’ molecular weight and primary structure determination. (10 hours classroom lesson) Topics: Light scattering and SEC-MALLS. Mass Spectrometry. Proteomic analysis.
Block 4: Proteins’ conformation determination (6 hours classroom lesson) Topics: Uv-VIS spectroscopy, fluorescence, FRET and fluorophores.
Block 5: Enzymes’ immobilization (2 hours classroom lesson) Topics: Main immobilization techniques and support materials. Bioreactors.
Block 6: Enzymes and sustainability (6 hours classroom lesson) Topics: The biorefinery concept, biomass definition. Enzymes and biofuels.
Block 7: Enzymes for environment (6 hours classroom lesson) Topics: Enzymes for environmental monitoring (biosensors) and for bioremediation.
The course includes classroom lectures and one laboratory session.
Slides published on the e-learning site of the Department of Science and Technology
"Lehninger Principles of Biochemistry", David L. Nelson, Michael M. Cox, Ed. Zanichelli
The final exam consists of an oral examination to verify the knowledge of metabolic pathways and their regulation, the knowledge of the structure and function of biomolecules.
The students’ skills will be verified by discussion of at least three of the seven blocks, that will be selected by the teacher during the examination.