Acquisition of knowledge of main analytical techniques for the investigation of proteins’ structure and function; ability to choose the most suitable techniques to implement sustainable processes and environmental monitoring.
Ability to disseminate, to not expert audience, what are the environmental issues that could be managed by means of biochemical techniques.
Ability 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 biochemical techniques and their potential spectrum of application.
The knowledge of the following topics, derived by the “Biochemistry” course, are required:
The levels of Proteins structure
Biological function of proteins
Enzymes as biological catalysts
Enzyme unit: definition and its determination
The interaction between electromagnetic radiation and matter: the spectroscopy techniques
Proteins’ concentration determination
Principles and application of chromatography
Principles and application of gel electrophoresis
General introduction to the applied biochemistry. Proteins’ investigation: chromatography, centrifugation, isoelectric focusing, immunochemistry.
Protein concentration and enzyme activity assays: more common techniques. Spectrofluorescence, luminometry.
Protein structure determination: light scattering and mass spectrometry. Proteome determination. Conformational studies: fluorescence, flurorophores and FRET.
Enzmyes: immobilization; application for sustainable processes, biofuels production (II and II generation bioethanol)
Environmental applications of enzymes: biosensors and bioremediation.
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 is based on classroom lectures.
Slides published on the e-learning site of the Department of Science and Technology
•Maccarrone “Metodologie biochimiche e biomolecolari. Strumenti e tecniche per il laboratorio”, Editore Zanichelli
The final exam consists of an oral examination to verify the knowledge of biochemical methods and their potential application for environmental monitoring. The students’ skills will be verified by discussion of three of the seven blocks, that will be selected by the teacher during the examination.