Chemistry and Biochemistry of Proteins, Nucleic Acides and Lipids.
This curricular unit consists of 8 introduction modules of theoretical-practical classes and laboratorial classes and a project to be intensively developed in four weeks. For the introduction modules, students are provided with summary information of the theme, the technology to be used and 2 or 3 key-words to be used in an internet research that must be carried out before the class. The project consists of the elaboration and autonomous implementation of a protocol for the purification of an enzyme that will be drawn from a previously presented list.
The main objective of this curricular unit is the acquisition of technical and practical knowledge of extraction, purification and characterization of enzymes.
Students are expected to acquire essential knowledge of the diversity of enzymatic activities and the physical-chemical characterization of biomolecules.
Students are also expected to:
-apply the acquired knowledge in the design and assessment of the processes of proteins’ purification and in the characterization of enzymatic activity.
-enhance their skills of organization, planning, and critical thinking in the laboratory
-enhance their skills in processing, analysis, presentation and discussion of experimental data.
1.Research of enzymatic activity in raw extracts. Enzyme assay design
2.Exploration of biomolecules’ physical-chemical properties in the process of separation
2.1.Molecular Exclusion Chromatography: principles and applications
2.2.Ion Exchange Chromatography.Principles. Anionic and cationic exchangers
2.3.Affinity Chromatography.Principles. Affinity, pseudo-affinity
2.4.Hydrophobic Interaction Chromatography. Principles and applications
2.4.1.FPLC and HPLC. Applications
2.5.Native electrophoresis and electrophorese in the presence of dissociating substances. Two-dimensional electrophoresis. Applications
2.6.Electrotransference, immunodetection and immunoprecipitation. Principles and applications
3.Detection and analysis of chemical modifications in proteins
4.Characterization and enzymatic activity. Determination of kinetic parameters Activation and Inhibition
5.Design and implementation of enzyme purification protocols. Evaluation of efficiency of the purification protocol
Paula Cristina Veríssimo Pires
Presentation and discussion of experimental results: 25.0%
Report of experimental results: 25.0%
R.K. Scopes ( 1994) Protein Purification: Principles and practice, 3rd ed., Springer-Verlag, New York.
Base de dados de enzimas.* :BRENDA
*consulta directa para servidores associados a establecimentos do ensino superior
Base de Dados de proteases: MEROPS
Base de dados de estrutura molecular de proteinas: PDB Protein Data Bank