67174 - General, Applied and Nutritional Biochemistry

Learning outcomes

At the end of the course the student has acquired the basic knowledge of biochemistry and molecular biology. In particular, the student is able to understand the structure and function of the main molecules of biological interest. The teaching also provides the basic knowledge of the techniques used in biochemical analyses and in particular the methods to separate and isolate the main

biological molecules, to reveal their functional qualities and to measure their concentration. The course includes a 24-hour lab focused to the practical aspects of the biochemical techniques treated during classes. Finally  the student  knows the basics about the biochemical and nutritional role of macronutrients, micronutrients and nutraceutical components in foods commonly used in human nutrition, and is able to understand the rational use of the main dietary supplements

Course contents

General Biochemistry (1A) 3 CFU–  Dr. Marco Malaguti

The molecules of life: properties of biomolecules. Energy and living systems. Covalent and non covalent bonds. Role of weak bonds in biological molecules.

Amino acids and proteins: Peptide bonds and protein sequence. Secondary structure. Motifs and structural and functional domains in the tertiary structure. Quaternary structure. Structure of myoglobin and haemoglobin. The heme group. Oxygen binding and cooperativity. Competitive inhibitors and allosteric ligands. Hemoglobin variants (embryonic, fetal and pathologic mutant forms).

Enzymes: Classification and general properties of enzymes. Activation energy. Catalytic mechanisms. Cofactors and prosthetic groups.

Kinetics of enzymatic reactions: Significance of kinetic parameters (Km, Vmax). The Michaelis-Menten equation. pH and temperature effect on enzyme activity. Enzyme inhibitors. Control of enzyme activity: allosteric control and covalent modifications.

Carbohydrates: monosaccharides, disaccharides, polysaccharides and glycoproteins.

Lipids and biomembranes: Fatty acids, triacylglycerols, glycerophospholipids, sphingolipids and steroids. Bilayers. Membrane integral and peripheral proteins. Structural organization and basic functions of biomembranes. Fluidity of membranes. The fluid mosaic model and lipid rafts. Transport across cell membranes, kinetics and thermodynamics of transport. Passive diffusion, uniporter catalyzed transport, active transport by ATP-powered pumps, cotransport by simporters and antiporters.

The central dogma of molecular biology. Nucleotide, nucleoside and base. Phosphodiester bond and nucleotide sequences. Base pairs and structure of double strand DNA.

Replication and enzymes involved in the process. Bacteria replicon.

Ribosomal, messanger and transfer RNAs. Structures of RNA. Procaryotic transcription. Procaryotic RNA polymerases. RNA polymerase subunits: sigma factor.

Hint of gene expression regulation. Genetic code: codon and anticodon.

Structure and function of tRNAs. Aminoacil-tRNA sinthetases. Ribosome structure. Translation mechanisms.

General Biochemistry (1B) 3 CFU– Dr. Francesco Vieceli Della Sega

Introduction to metabolic pathways. Thermodynamics of metabolic pathways. Control of metabolic flux. ATP and “high-energy” compounds. Coupled reactions. Redox reactions.

Cellular energetics and glucose metabolism. Oxidation of glucose: the reactions of glycolisis. The anaerobic metabolism of pyruvate: fermentations. Regulation of glycolisis. The pentose phosphate pathway. Glycogen metabolism and gluconeogenesis: glycogen breakdown, synthesis and control. Gluconeogenesis: reactions and regulation.

The citric acid cycle. Mitochondrial oxidation of pyruvate and synthesis of acetyl-Coenzyme A. Reactions of the citric acid cycle and its regulation. Reactions related to the citric acid cycle.

Mitochondrial electron transport and oxidative phosphorylation. Electron transporters in mitochondria. Oxidative phosphorylation: the proton-motive force and the chemiosmotic theory. ATP synthase. Uncoupling and inhibition of electron transport. Control of oxidative metabolism.

Oxidation of fatty acids. Fatty acid oxidation. Ketone bodies. Fatty acid biosynthesis: reactions and regulation. Regulation of fatty acid metabolism. Synthesis of triacilglycerols and glycerophospolipids.

Catabolism of amino acids. The aminotransferase reactions. Amino acid deamination. Protein degradation. Cell signalling. Signal transduction mechanisms, the cAMP and phosphatidylinositol pathways. Receptor tyrosine kinases

General Biochemistry (2) 2 CFU– Prof. Paolo Neyroz

The course includes a 24-hour practical class focused to the experimental aspects of the biochemical techniques treated during classes.

 

Applied Biochemistry (3) 3CFU Prof. Silvana Hrelia

General principles of biochemical investigations: methods for investigating metabolism, in vivo models, in vitro models, primary cell cultures and cell lines.

Spectrophotometric methods: ultraviolet and visible light spectroscopy (principles, instrumentation, applications).

Spectrofluorimetry: principles, instrumentation, applications. Fluorescent probes.

Radioisotopic techniques: the nature of radioactivity, interaction with matter, detection and measurement, instrumentation, safety aspects, application of radioisotopes in the biological sciences.

Electrophoretic techniques: general principles, instrumentation, support media, electrophoresis of plasma proteins.

Immunochemical techniques: antibodies and their production, antisera, immunoprecipitation and immunodiffusion, RIA, IRMA. Labelling with enzymes: EMIT, ELISA, PEIA. Pregnancy test.

Basic techniques in molecular biology: manipulation of nucleic acids, the polymerase chain reaction, nucleic acid blotting methods, protein blotting methods. Gene expression analysis: the DNA microarray technique. Large scale protein expression analysis: the proteomic approach.

 

Nutritional Biochemistry (4) 3CFU Dr. Emanuela Leoncini

 

 

Food and Nutrition: Definition of Food Science and Nutrition Science. Food quality/safety and its determinants from farm to fork. The role of consumers and the new "from fork to farm" assumption.

Nutritional standards and guidelines: Formulation of dietary guidelines. Body mass index (BMI) and biochemical evaluations, RDA, food pyramids. Categories of nutrients, macronutrients and micronutrients. From nutrients to foods. The seven classes of foods.

Nutrition and metabolism of carbohydrates: food sources, digestion, absorption, metabolism and function. Lactose intolerance. Galactosemia. Foods and glucose tolerance, glycemic index, nutrition and diabetes. Dietary fiber.

Nutrition and metabolism of fats: food sources, absorption, metabolism and function. Saturated, monounsaturated and polyunsaturated fatty acids. Trans fatty acids. N-6 and n-3 fatty acids: biosynthesis, metabolic functions, RDA, nutritional sources. Cholesterol, nutrition and control of serum cholesterol concentration. Lipoproteins.

Nutrition and metabolism of proteins: essential aminoacids, food sources of proteins and protein quality, digestion, absorption, metabolism and function. Nitrogen balance. Protein requirements.

Water: endogenous and exogenous sources. Water requirement. Water homeostasis in the body.

Ethanol metabolism, diet and alcoholism. Coffee, tea, cocoa and chocolate.

Nutrition and metabolism of vitamins: nomenclature, sources and properties. Vitamin A, D, K and E: absorption, metabolism, storage, functions, requirements, deficiency and toxicity. Vitamin C: function, metabolism, requirements, deficiency and toxicity. The vitamin B complex: function, metabolism, requirements, deficiency and toxicity.

Nutrition and metabolism of the major minerals and of trace elements: absorption, metabolism, excretion, storage, requirements, deficiency and toxicity.

Functional foods and Nutraceuticals: Oxidative stress and reactive oxygen and nitrogen species. Intracellular and extracellular antioxidants. Functional foods and nutraceutical compounds in the prevention of chronic degenerative diseases.

Dietary supplements: definition and legal regulation in the EU. Reasons for taking supplements. Vitamin and mineral supplements: judging the adequacy of micronutrient intakes. Protein supplements. Branched chain aminoacids and creatine: rationale and evidence for the use of supplements.

 

 

Readings/Bibliography

General Biochemistry

D.L.Nelson and M.M. Cox - Lehninger principles of Biochemistry - 4th Edition, Freeman 2004

D.Voet, J.G.Voet and C.W.Pratt - Fundamentals of Biochemistry - 2nd Edition, Wiley 2006

R. H. Garrett , C. M. Grisham –Biochemistry – 4th edition, Mary Finch, 2010

J.M. Berg, J.L. Tymoczko, L. Stayer-Biochemistry -7th edition, Freeman 2012

 

Applied Biochemistry

 

K. WILSON, J. WALKER. Principles and techniques of practical biochemistry. Cambridge University Press, Cambridge 2005

 

Nutritional Biochemistry

 

M.C. Linder (ed.) Nutritional Biochemistry and metabolism with clinical application. Appleton & Lange, Norwalk, Connecticut (USA)

Teaching methods

Theoretical lessons. A practical class (2 ECTS) will deal with laboratory experiments and clinical applications (Biological laboratory, Via Bastioni Settentrionali, 45 Rimini).

Assessment methods

The examination at the end of the course aims to assess the achievement of learning objectives:

- to know the structure and function of the main biological macromolecules and the basis of enzymology, to know the cellular bioenergetics, the main metabolic pathways and the mechanisms underlying their regulation and signal transduction, to know the fundamental concepts of molecular biology, the basic molecular mechanisms of living organisms and their regulation.

- to know the main techniques used in biological and biochemical studies,  to know and apply the main analysis protocols used in biochemistry laboratories

- to know the macro and micronutrients in food and their metabolic role,  to know the role of functional foods and bioactive nutraceutical compounds in the prevention of chronic/degenerative diseases,  to know the rationale and evidence to use nutritional supplements in sedentary people and in physical activity.

Final examination consists of an oral examination on all the modules, which can be broken down into moments of verification separated (General Biochemistry, Applied Biochemistry, Nutritional Biochemistry). The single module mark is defined by an oral test with two questions aimed at evaluating the acquired knowledge. Over the course a written test in progress can be set up "one-off" for the single modules consisting of open-ended questions that if overcome with positive vote (>18) will count as the acquisition of knowledge and will contribute for to the final vote.  The final vote of the course will be the weighted average of the marks obtained in the two modules.

 

Teaching tools

PC, overhead projector, biological laboratory

Office hours

See the website of Silvana Hrelia

See the website of Marco Malaguti

See the website of Francesco Vieceli Dalla Sega

See the website of Paolo Neyroz

See the website of Emanuela Leoncini