LEARNING OUTCOMES

The teaching of Biochemistry aims to provide the knowledge for understand the mechanisms that are the basis of life by describing  the structure and function of biomolecules. The principles of bioenergetics that rule the biochemical transformations will allow to clarify the metabolic changes that undergo biomolecules and their interactions both at the molecular and cellular level. Finally, we will address the biochemical aspects related to genetic information and how it is translated into amino acid sequences. The teaching of applied biochemistry provides general approaches to biochemical testing.

AIMS AND LEARNING OUTCOMES

The study of Biochemistry has as objective the study of the structure and function of the cell to a better understanding of the vital processes occurring in living beings with particular attention to the interrelations of catabolic and anabolic processes and their regulation mechanisms. Understand, in fact biochemical processes is a prerequisite for any professional profile in the biological sciences. During the course we will delve into the structure, function and metabolism of carbohydrates, lipids and proteins with emphasis on enzymatic catalysis to its regulation and its control. Then It will be introduced the concepts of Molecular Biology with a focus on the processes of DNA replication, RNA transcription and protein synthesis via translation. The final part of the course will be dedicated to Applied Biochemistry with the deepening of the main techniques used in a biochemical laboratory with particular attention to those used in protein purification processes. The topics covered in the course are the chemistry of biological macromolecules with references to physicochemical, thermodynamics and bioenergetics.

During the course, students must achieve certain objectives:

• to acquire the concept of the relationship between structure and function of biomolecules, especially proteins
• to acquire the concept of relationship between structure and function of enzymes. Particular attention should be given to enzyme catalysis.
• to acquire the concept of relationship structure and function of carbohydrates and lipids
• to understand the metabolic pathways of catabolic type used to obtain energy from the degradation of biomolecules
• knowing the type of anabolic pathways for carbohydrates, lipids, amino acids and nucleotides, the components necessary for cell structure and function
• to know the basics of molecular biology: DNA as the genetic material, the mechanisms of replication, transcription, translation and regulation.
• to learn from the theoretical point of view of the fundamental principles of the main techniques used in a biochemical laboratory, paying particular attention to the techniques used in the purification of proteins.

TEACHING METHODS

Lectures and discretionary laboratory

SYLLABUS/CONTENT

Amino acids and proteins Classification and chemical properties of amino acids. Primary, secondary and tertiary protein structure: examples. quaternary structure. Hemoglobin and myoglobin: structure and function. Pathological hemoglobins. Catalysis and enzyme kinetics: factors and laws that regulate it. Enzyme inhibition, allosterism and regulating the activity of enzymes.

Metabolism and energy problems connected Anabolism and catabolism, synthesis and degradation of high-energy bonds.

Glucose metabolism Structure of main mono, di and polysaccharides. Main glucose utilization pathways. Glycolysis: enzymatic steps, co-enzymes and vitamins concerned, energy balance and regulation. Hexose monophosphate shunt and its biological importance. Reduced glutathione: structure and functions. Interconversion of phosphorylated sugars. Synthesis and degradation of glycogen and hormonal regulation (adrenaline, glucagon and insulin). Metabolism of fructose, mannose and galactose. Gluconeogenesis: specific enzymatic steps and regulation. The oxidative decarboxylation of pyruvic acid: vitamin co-enzymes and enzymes involved. The citric acid cycle: enzymatic steps, energy balance and regulation.

Lipid Metabolism Oxidative demolition of the main classes of fatty acids (odd and even, saturated and unsaturated): enzymatic steps and energy efficiency. Formation of ketone bodies in normal and pathological conditions. Synthesis of fatty acids: Fatty acid synthase complex, lengthening process and unsaturation. Synthesis and degradation of triacylglycerols, phospholipids, sphingolipids. Arachidonic acid metabolism. Cholesterol: structure, function, enzymatic steps of the synthesis and regulation. Synthesis of compounds derived from cholesterol and isoprenoid.

Bioenergetics Oxidative Phosphorylation: organization and localization of protein complexes in mitochondria Electron transport, ATP synthesis and proton pumps. Photosynthesis: light and dark phase. Photorespiration.

Nitrogen metabolism Metabolism of amino acids. Transamination and oxidative deamination. Stages enzyme of the urea cycle. Oxidative degradation of the carbon skeleton of amino acids. Biosynthetic processes related to amino acids. Ex-novo synthesis, recovery streets and demolition of purine and pyrimidine nucleotides.

The biological information Structure of nucleic acids. DNA replication. Transcription. RNA maturation. Genetic code. Protein synthesis. Post-translational modifications, transport and demolition of proteins. Regulation of gene expression in prokaryotes: lac operon.

Applied Biochemistry General Approach to biochemical research. Working in lab: Good Laboratory Practices, basic instrumentation, solutions. Protein purification: separation techniques (cell breakage methods, centrifugation techniques, fractional precipitation, chromatographic techniques) and analytical techniques (electrophoresis, spectrophotometric and radioisotopic techniques, immunochemical techniques, cytofluorimetric techniques). Determination of primary structure and molecular weight of proteins by Edman degradation and mass spectrometry.

RECOMMENDED READING/BIBLIOGRAPHY

• Nelson D.L e Cox M.M., I Principi di Biochimica di  Lehninger, Zanichelli ed.
• Berg J., Tymoczko J.L. e Stryer L., Biochimica, Zanichelli ed.
• Garret R.H. e Grisham C.M., Biochimica con aspetti molecolari della Biologia cellulare, Zanichelli ed.
• Voet D., Voet J.G. e Pratt C.W., Fondamenti di Biochimica, Zanichelli ed.
• Garret R.H. e Grisham C.M., Biochimica, Piccin ed.
• Stoppini M., Bellotti V., Biochimica Applicata, EdiSES Ed.
• Bonaccorsi Di Patti M.C., Contestabile R., Di Salvo M.L., Metodologie Biochimiche, Zanichelli Ed.
• Maccarrone M, Metodologie Biochimiche e Biomolecolari, Zanichelli Ed.