Cell biology and technology

APPLIED BIOCHEMISTRY

Course objectives

General skills The course aims to provide students with the biochemical bases to understand the most advanced biotechnological applications of enzymes, proteins and complex multienzymatic systems. The biochemical principles of methods for the study of protein-protein interactions will also be illustrated. Furthermore, aspects of metal metabolism in procaryotes and eukaryotes will be illustrated. Specific skills A) knowledge and understanding - knowledge of the main biotechnological applications of enzymes,proteinsandcomplex multienzymatic systems; - knowledge of the main biochemical techniques for the study of protein-protein interactions - knowledge of the strategies required for protein and enzyme production and engineering B) ability to apply knowledge and understanding - exploiting the knowledge of biochemical techniques to investigate the applications of enzymes and proteins in the field of biotechnology - understanding and evaluating the impact of structural modifications of biological macromolecules on their biological function; C) Making judgements - critical thinking through the study of examples of biotechnological applications of proteins and enzymes taken from the scientific literature - learning by questioning D) Communication skills -ability to communicate what has been learned during the oral exam E) Learning skills - learning the specific terminology - ability to make the logical connections between the topics covered - ability to identify the most relevant topics

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MARIA CARMELA BONACCORSI DI PATTI Lecturers' profile

Program - Frequency - Exams

Course program

The course is composed of sections that deal with different aspects of biochemical applications of enzymes and proteins. Biotechnological applications of enzymes. Properties of enzymes: chemo-, regio- and stereo-selectivity. Reversible, irreversible and ‘suicide’ enzyme inhibition: practical examples (inhibition of the biosynthesis of folic coenzymes, inhibition of thimidylate synthase and beta-lactamase). Main techniques of cell and enzyme immobilization. Use of proteases, lipases and penicillin acylase for production of biotechnologically relevant compounds. Production of recombinant enzymes and proteins: methods and strategies. Enzyme engineering: site-specific mutagenesis, in vitro evolution. Multienzymatic systems for biosynthesis of bioactive secondary metabolites. Non-ribosomal biosynthesis of peptides. Enzyme mechanism of peptide synthetases. Examples (gramicidin, surfactin, penicillins and cefalosporins). Biosynthesis of polyketides. Mechanism of polyketide formation and analogies with fatty acid biosynthesis. Examples (erythromycin). Methods for the study of protein-protein interactions. Biochemical methods to identify interacting proteins: affinity chromatography, ligand blot and use of cross-linkers. Systems based on split-proteins. Biochemical methods for the quantitative study of protein-protein interactions. Biosensors. The ‘bio’ and the ‘sensor’ components. Examples of applications for biomedical, food and pesticide analyses. Metallobiochemistry. Metalloproteins and aspects of metabolism of metals. Mechanisms of copper and iron homeostasis in prokaryotes and eukaryotes. Guided activities in the laboratory.

Prerequisites

No formal propedeuticity is required. However, basic knowledge of biochemistry, molecular biology as well as of cellular biology is necessary.

Books

Text books and other materials are promptly published at the educational site Elearning2 at http://elearning2.uniroma1.it

Teaching mode

Classroom lectures and guided activities in the laboratory

Frequency

Lectures and guided activities are not mandatory.

Exam mode

The aim of the exam is to test the level of knowledge and deepening of the topics of the course program and the reasoning skills developed by the student on issues related to applied biochemistry. The assessment is expressed in thirtieths (minimum grade 18). The exam is oral and the answers to the questions are evaluated both for the content and for the appropriate scientific language. The student can choose to take an oral exam with open questions or produce an oral presentation of a scientific paper regarding a topic of the course program.

Lesson mode