FA/0260 - BIOCHEMISTRY AND BIOCHEMISTRY LABORATORY
Academic Year 2022/2023
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TIZIANA CABRAS (Tit.)
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|[60/76] BIOTECHNOLOGY||[76/20 - Ord. 2018] Farmaceutico||12||108|
The course of Biochemistry for the Class Biotechnology is aimed to the acquisition of fundamental information and concepts in structural and functional biochemistry. Therefore, it gives basic biochemical knowledge for a first-level degree in Biology, and moreover it is preparatory to further learning courses, implying the knowledge of such fundamentals.
The Laboratory Module consolidates the manual skills already acquired in previous courses and adds new ones, especially the ability to manipulate protein and enzyme solutions, and to carry out the most common analytical determinations on them.
Knowledge and Understanding:
Understanding the Life molecular logic and the primordial abiotic chemistry.
Knowing structures and properties of the main biological compounds such as proteins, glucides, and lipids. Being able to graphically represent their molecules.
Understanding the structural and functional features of those compounds and their localization in cells and tissues.
Knowing and understanding structure/function relationship of the proteins, and their modulation: the case of respiratory proteins.
Knowing the modes of production, storage, use of metabolic energy, and understanding the related thermodynamics.
Knowing the main metabolic pathways and their molecular tools: enzymes, coenzymes, cofactors, effectors, inhibitors.
Understanding metabolic homeostasis and the general integration of metabolic pathways.
Knowing the main laboratory techniques.
Understanding the theoretical and practical bases for the study of macromolecules.
Applying Knowledge and Understanding:
The students will be able to:
- take advantage from structure formulas getting the main reaction mechanisms applied to biochemistry for both catalysis and metabolic sequences;
- correctly put the various metabolic pathways within the frame of physiological and eventually pathological mechanisms to get conclusions about material and energy balances;
- apply the acquired knowledge to the study of Molecular Biology, Genetics, Physiology, Microbiology.
- apply the laboratory practices also to contexts other than biochemical ones;
- apply the theoretical-practical knowledge acquired for standard analysis of biological matrices.
Students will be able to:
- distinguish among main and secondary topics, finding the logics lying under the study of Biochemistry, and avoiding a memory-based study;
- autonomously find and explain retrohinibition, cycles, and futile cycles, anaplerotic reactions;
- individuate the basic concepts to apply them to other disciplines along the Biology course.
- be able to organize a laboratory experiment by choosing the most suitable technique for the type of analysis to be carried out;
- correctly interpret the laboratory analytical data obtained to draw correct and consistent conclusions.
The students will be able to discuss the topics of the course by using the language and vocabulary typical for Biochemistry. They will interact with the teacher arguing the main points of the study program with the required detail. Owing to the expert tutorage they will also positively interact with their colleagues, possibly forming study groups.
Student/teacher interaction, auto-evaluation tests, slides and booklets will give the students the required tools for profitable and aware learning.
The students must have a sound basic knowledge of both Inorganic and Organic Chemistry.
It is mandatory to have taken the exams considered preparatory.
The course divided in two learning units: 1) Structural Biochemistry, and 2) Functional Biochemistry.
Biochemistry (6 CFU; 48 hours):
- Aminoacids and peptides, fibrous proteins. Aminoacids: definition and classification, main physicochemical features. Aminoacids as electrolytes. Essential aminoacids. Peptides. Protein classification and structural levels. The main fibrous proteins.
-Globular proteins. Main physicochemical features. Globular proteins as amphoionic polyelectrolytes. Glycoproteins. Immunoglobulins. Stability and denaturation of globular proteins.
-Glucides. Definition and classification, main physicochemical features. Anomerism, mutarotation, glycosides. Mono-, oligo-, and polysaccharides.
- Nucleotides and nucleic acids. The purinic and pyrimidinic bases. The nucleotides.
-Lipids: definition, classification, main physicochemical features. Simple and complex lipids, phospholipids, sphingolipids. Isoprenoids.
- Porphyrins, metalloporphyrins, heme, hemoproteins and their main physicochemical features. Myoglobin and hemoglobins, oxygenation and deoxygenation, cooperativity and its logical models.
-Enzyme kinetic. Catalysis and biocatalysis, enzymes. Molecularity and reaction order of enzymatic reactions. Michaelis and Menten hypothesis and its developments. Enzyme activity inhibition and modulation.
-Enzymatic catalysis: physicochemical background of affinity and catalysis. Theory of the activated complex and enzymatic catalysis. Biocatalytic mechanisms.
-Vitamins and coenzymes: Definition and classification. Hydrosoluble vitamins, coenzymes and cofactors.
-Metabolism and energy metabolism: Definition and concepts: metabolism, anabolism, catabolism, biosynthesis, biodegradation. Aerobic and anaerobic metabolism. Metabolic paths and cycles, energy production, transportation, storage of metabolic energy: ATP and analogues.
- Glucidic metabolism: General concepts, glycolysis, lactic and alcoholic fermentations. Citrate and glyoxylate cycles.
-Lipid metabolism: General concepts, beta-oxidation, particular cases, AcetylCoa fate, ketogenesis.
-Protein metabolism: General concepts, transamination, deamination, ammonia fate, ornithine cycle.
-Respiratory chain: General concepts, constituents, consequences and purposes.
-Oxidative phosphorylation: general concepts, ATP synthase and its mechanism, respiratory control, uncoupling agents.
Methods (60 hours):
Lecture syllabus (24 hours)
Purification of protein. Homogenate preparation. Fractional precipitation with ammonium sulfate, organic solvents and precipitation heat. Centrifugation. Dialysis. Ultrafiltration. 4 hours.
Chromatographic techniques: general principles. Ion-exchange, exclusion, adsorption and affinity chromatography. HPLC. 6 hours.
Spectrophotometry UV/VIS: principles, instruments and applications. Lambert-Beer law. Fluorescence analysis: principles, instruments and applications. 6 hours.
Electrophoretic Techniques: general principles. PAGE and SDS-PAGE. Methods of detection and quantitative assessments. Isoelectric focusing (IEF). Western blot. 8 hours.
Laboratory syllabus (36 hours)
Determination of protein concentration by using Bradford method. 5 hours
SDS-PAGE and determination of the molecular weight of a protein. 6 hours
Enzymatic activity on PAGE. 5 hours
Zymogram with SDS-PAGE. 5 hours
Enzymatic activities using spectrophotometric assays and calculation of kinetic parameters (Km e IC50). Chromogen substrates. 10 hours
Assay for the evaluation of antioxidant activity of a solution. 5 hours.
Teaching will be delivered in person. The lessons can be integrated with audiovisual materials and streaming.
The course starts in October and ends in January. Are provided:
- 72 hours of lectures (including 48 hours for the Biochemistry module and 24 hours for the laboratory module) and 24 hours in the laboratory.
Biochemistry module: 20 hours on the structure of biomolecules and their biological properties (sugars, lipids, nucleic acids, amino acids and proteins, hemoproteins); 10 hours on enzymology and the structure and function of coenzymes; 18 hours on metabolism.
Laboratory Module: 24 hours of lecture including 4 hours about protein purification procedures, 6 hours about chromatographic techniques, 6 hours about spectrophotometric techniques and 8 hours about electrophoretic techniques; 36 hours of laboratory activities including 20 hours about spectrophotometric techniques and 16 hours about electrophoretic techniques.
The teacher will provide individual explanations during reception hours by appointment, and / or with the help of the available computer media.
To assist in the study, the teacher makes the slides of the lessons available, and provides exercise books and copies of previous written tests.
Verification of learning
Biochemistry: Written and oral exam. The written biochemistry test can be evaluated for the purposes of the final exam and will take place on the following topics:
- Structure and acid-base properties of amino acids and peptides; structure of proteins, carbohydrates, lipids, nucleotides and ac. nucleic; biological membranes. Properties of respiratory proteins. Interpretation of the bond curves.
The assessment of the test is not considered definitive until the student has demonstrated in the final oral exam that he has recovered all the topics on which he has taken insufficiencies.
The student can repeat the written test on the day of the exam or bring the topics back to the oral exam. For non-attending students, on the day of the exam, there is a written biochemistry test that will precede the oral exam.
Laboratory Module: oral exam.
For both modules, will be verified:
a) acquisition of related, general and basic, detailed and specific notions
b) logical integration of the acquired notions, their understanding and interpretation
c) ability to develop the knowledge acquired for study, work or profession
d) ability to carry out, understand and explain laboratory experiences
The final grade takes into account:
a) Quality of knowledge, skills, competences possessed and / or manifested.
b) Exhibition method:
c) Logical capacity and consequentiality of the contents
d) Manual and practical skills
e) Ability to connect different topics by taking care of the structure, organization and logical connections of the expository discourse
f) Ability to synthesize also through the use of the symbolism of the material and the graphic expression of notions and concepts.
g) Relational quality and willingness to interact with the teacher during the interview.
h) critical spirit; self-assessment skills; ability to operate also in a group.
The judgment can be:
• 18-20. The candidate demonstrates few acquired notions, superficial level, many gaps. Modest expressive skills but still sufficient to support a coherent dialogue; logical skills and consequentiality in connecting elementary level arguments; poor synthesis skills and rather poor graphic expression skills; poor interaction with the teacher during the interview
• 21-23. The candidate demonstrates a fair acquisition of notions, but little depth, few gaps; expressive skills more than sufficient to support a coherent dialogue; acceptable command of scientific language; logical skills and consequentiality in the connection of topics of moderate complexity; more than sufficient capacity for synthesis and acceptable graphic expression capacity
• 24-26. The candidate demonstrates a rather broad knowledge base, moderate in-depth study, with small gaps; satisfactory expressive skills and significant command of scientific language; dialogic ability and critical spirit well noticeable; good synthesis skills and more than acceptable graphic expression skills
• 27-29. The candidate demonstrates a very extensive, well-studied knowledge base, marginal gaps; remarkable expressive skills and high command of scientific language; remarkable dialogic ability, good competence and relevant aptitude for logical synthesis; high capacity for synthesis and graphic expression.
• 30. The candidate shows a very extensive and in-depth knowledge base, high expressive skills and mastery of scientific language; excellent dialogue skills, strong aptitude for making connections between different topics; excellent ability to synthesize and great familiarity with graphic expression
Honors are attributed to candidates clearly above the average and whose possible notional, expressive, conceptual or logical limits are, on the whole, completely irrelevant.
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- L. Pollegioni, Fondamenti di Biochimica, EdiSES
- Campbell, Farrell, McDougal, Biochimica, EdiSES
- Voet-Voet-Pratt; Fondamenti di Biochimica; Zanichelli
- Jeremy M. Berg, John L. Tymozcko, Gregory J. Gatto, Lubert Stryer, Biochimica; Zanichelli
- Garrett-Grisham; Principi di Biochimica; Piccin
- Mathews-Van Holden-Appling- Anthony Cahill; Biochimica; Piccin
Bonaccorsi di Patti et al., Metodologie Biochimiche, Zanichelli;
Stoppini, Bellotti. Biochimica Applicata. EdiSES
- Cabras Tiziana
Department: Life and Environment Sciences
Adress: Cittadella Universitaria Monserrato
Student will be received every working day by appointment.
- Francesca Pintus, Department of Life and Environment Sciences, Biomedical Section, Cittadella Universitaria S.S 554 km 0.700 bivio per Sestu, 09042 Monserrato.
Student will be received every working day by appointment.